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Salidroside inhibits migration and invasion of human fibrosarcoma HT1083 cells.

ARTICLE INFO

Keywords: Salidroside invasion Metastasis HT1080 cells Reactive oxygen species

ABSTRACT

Oxidative stress plays an important role in tumorigenesis and metastasis. Salidroside, a phenylpropanoid glycoside isolated from Rhodiola roses L, shows potent antioxidant property. Here we investigated the inhibitory effects of salidroside on tumor metastasis in human fibrosarcoma HT1080 cells in vitro. The results indicated that salidroside significantly reduced wound closure areas of HT1080 cells, inhibited HT1080 cells invasion into Matrigel-coated membranes, suppressed matrix metalloproteinases (MMP-2 and MMP-9) activity, and increased tissue inhibitor of metalloproteinase-2 (TIMP-2) expression in a dose-dependent manner in HT1080 cells. Salidroside treatment upregulated the E-cadherin expression, while downregulated the expression of [beta]1-integrin. As an antioxidant, salidroside inhibited the intracellular reactive oxygen species (ROS) formation in a dose-dependent manner. The results also showed that salidroside could inhibit the activation of protein kinase C (PKC) and the phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) in a dose-dependent manner. In conclusion, these results suggest that salidroside inhibits tumor cells metastasis, which may due to its interfere in the intracellular excess ROS thereby down-regulated the ROS-PKC-ERK1/2 signaling pathway.

Introduction

Cancer has been the leading cause of global deaths, the mortality of cancer still rises every year (Hanahan and Weinberg 2000). Malignant tumor cells often metastasize to distant organs, and metastatic tumor recurrence can occur many years after treatment of the primary tumor, then leading to the death of the patient. Although metastasis is a major target of cancer therapy, the high death rate from cancer clearly indicates the difficulty in preventing or inhibiting it. Most anti-cancer agents, which rely on the differences in the proliferation rate of normal and tumor cells, have little effect on micrometastases because tumor cells distributed to distant organs often remain dormant. Other therapeutic methods, such as surgical removal or irradiation therapy, are not generally suitable for treatment of micrometastases. Previous studies in clinical and experimental settings have reported that the removal of a primary tumor aggravates the growth of micrometastases (Goodison et al. 2003). There are no effective therapeutic approaches to treat cancer recurrence and metastasis until now.

Difficulties in the treatment of cancer are attributed mainly to the complex nature of metastasis. Tumor metastasis consists of a series of steps: angiogenesis of endothelial cells (ECs) in solid tumor, dissociation and intravasation of cells from a primary tumor into the circulation, arrest in small vessels in down-stream organs, adhesion and extravasation into surrounding tissues, proliferation (Bogenrieder and Herlyn 2003; Gupta and Massague 2006). In these complex steps cancer cells are not the sole players, but they interact with extracellular matrix, surrounding tissues, blood cells, and endothelial cells. Previous studies have demonstrated that interaction of cancer cells with these components results in changes not only in cancer cells but also non-cancerous cells, which leads to the altered expression of various genes closely involved in tumor metastasis. Therefore, the complex nature of tumor metastasis cannot always be attributed to the abnormalities of cancer cells (Nishikawa 2008).

ROS have been identified as a kind of important messengers involved in the transduction of several signaling pathways (Reth 2002; Forman et al. 2002), gene expression, and cell proliferatio (Sauer et al. 2001). Recently, the involvement of ROS signaling in tumor metastasis was highlighted (Storz 2005; Radisky et al. 2005). One of the important down-stream signal cascades regulated by ROS is mitogen activated protein kinase (MAPK) including ERK, JNK and p-38. Studies have shown that oxidative activation and/or inactivation of PKC and PTP (Shackelford et al. 2000; Chiarugi 2005; Lee and Esselman 2002), respectively, by ROS result in MAPK activation followed by activation of various transcriptional factors including SMAD, AP-1, Ets-1 and Snail (Lin et al. 2004; Lo et al.2005; Rhyu et al. 2005; Wu et al. 2008). Each transcriptional factor then regulates its target genes such as E-cadherin, MMPs, integrin and biglycan leading to epithelial-mesenchymal transition (EMT), migration and invasion (Wu 2006).

Salidroside (p-hydroxyphenethyl-[beta]-D-glucoside, Fig. 1), a phenylpropanoid glycoside extracted from Rhodiola rosea L. which has long been used as an adaptogen in traditional Tibetan medicine, has been reported to possess various pharmacological properties including antiaging, anticancer, neuroprotective, anti-inflammation, hepatoprotective and strong antioxidative activities (Mao et al. 2010; Kanupriya et al. 2005; Kucinskaite et al. 2004; Zhang et al. 2007; Cao et al. 2005). Nevertheless, no report has been issued on the interrelation of salidroside with tumor metastasis. The aim of this study was to evaluate its anti-metastasis effect in vitro and to reveal the anti-mechanism from ROS-PKC-ERK pathway.

Materials and methods

Materials

Salidroside (purity >99%) was purchased from National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Salidroside was dissolved in water and filtered through a 0.22[micro]M filter before use. 2,7-Dichlorofluorescin diacetate (DCFH-DA), 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide (MIT), DMEM medium were purchased from GIBCO Chemical Co. Fetal bovine serum (FBS) were purchased from Hangzhou Sijiging Biological Manufacture Co.,Ltd.(Hangzhou, China). TIMP ELISA Kit were purchased from SenXiong Biotech Co., Ltd. (Shanghai, China).[beta]1-Integrin monoclonal antibody and E-cadherin monoclonal antibody were obtained from Biosynthesis Biotechnology Co., Ltd. (Beijing, China). All other chemicals were of analytical grade and commercially available.

Cell culture

Human fibrosarcoma HT1080 cells were obtained from China Center for Type Culture Collection (CCTCC), Wuhan, China. HT1080 cells were maintained in DMEM medium supplemented with 10% fetal calf serum, 100 U/m1 penicillin and 100 U/m1 streptomycin in a humid atmosphere of 5% C[O.sub.2] and 95% air at 37[degrees]C.

MTT assay

The cytotoxic activity of salidroside was evaluated in 1-1T1080 cells using the MTT assay (Mosmann 1983). Cells were plated in 96-well plates (0.5 x [10.sup.5] cells/well for cells in 100 [micro]l of medium).100[micro]1 salidroside solution was added to each well at a final concentration of 10-100 [micro]mo1/1. The cells were exposed to the drug for 24 h. Thereafter, the medium was replaced by fresh medium (200 [micro]1) containing 0.5 mg/ml MTT. After 3 h incubation, the MITT formazan was dissolved in 150 ill DMSO, and the optical absorbance was measured at 570 nm with reference at 630 nm using a microplate reader (Thermo Varioskan Flash 3001, USA).

Trypan blue exclusion test

The lethality of salidroside on HT1080 cells was determined by the trypan blue exclusion test (Li et al. 2009). After 24 h incubation with salidroside from 10 to 100 [micro]mo1/1, HT1080 cells were removed from cultures and cells that excluded trypan blue were counted in a Neubauer chamber.

Scratch wound closure assay

For detection of HT1080 cells migration, cells were grown in 48-well plates, and a portion of cells monolayer was scraped away with a sterile disposable rubber policeman (Weis et al. 2002). The remaining cells were washed with medium and incubated with different concentrations (10, 20 and 40 [micro]mo1/1) of salidroside for 24 h. HT1080 migration into the denuded area was quantified with a computer-assisted microscope (ZEISS, M1C00266).

In vitro invasion assay

Invasion of HT1080 cells in vitro was measured by the invasion of cells through Matrigel-coated transwell inserts (Wu et al. 2009).Firstly, serum-free DMEM diluted Matrigel matrix (60 [micro]g/well) was put into the upper chamber of transwell filter (8 [micro]m pore size) and incubated for 4 h at 37[degrees]C for gelling. Then, cells were tiypsinized and seeded at 2 x [10.sup.5] per upper chamber in medium with or without addition of salidroside. The cells in the upper side of the insert membrane were rubbed with a cotton swab. Cells that had migrated to the underside of the insert membrane were fixed with methanol and dried. The cells were then stained for 20 min with hematoxylineosin staining solution, rinsed in deionized water, air-dried, and observed under a microscope equipped with a camera. For quantitation, the migrated cells on the underside were counted based on the number of the cells under a microscope at 200x magnification field. Eight fields per insert were scored and averaged.

Assay of intra-cellular reactive oxygen species

HT1080 cells incubated with salidroside (10, 20 and 40 [micro]mol/l) for 24 h were washed with serum-free and phenol red-free DMEM and loaded with 5 p.M 2,7-dichlorofluorescin diacetate (DCFH-DA).After incubation for 30 min in the dark, cells were washed with PBS twice, and observed with fluorescence microscopy (ZEISS, MIC00266). Then, cells were detached, and resuspended in 1 ml of PBS. Cellular ROS in 10,000 cells as a result of the oxidation of 2,7-dichlorofluorescin diacetate was measured (excitation, 470 nm; emission, 530 nm, Thermo Varioskan Flash 3001, USA) (Martinez et al.2008).

Measurement of MMP-2 and MMP-9 activity by zymography

HT1080 cells (300,000/1.5 ml) were initially grown in DMEM supplemented with 10% FBS in petridishes, washed with serumfree culture medium, and treated with increasing concentrations of saldroside (10, 20 and 40[micro]mo1/1) for 24 h in serum-free culture medium. Cells were cultured for 24 h and the medium were collected. The conditioned medium was then subjected to zymography on 7.5% SDS-PAGE copolymerized with 0.1% gelatin. Gel was washed in 2.5% Triton-X-100 for 30 min to remove SDS and was then incubated overnight in reaction buffer (50 mM Tris-HCI pH 7, 4.5 mM Ca[Cl.sub.2], 0.2 M NaCI). After incubation, the gel was stained with 0.5% coomassie brilliant blue in 30% methanol and 10% glacial acetic acid (Dutta et al. 2009). The bands were visualized by destaining the gel with water.

Assay of tissue inhibitors of meralloproteirtase-2 (TIMP-2) content

Tissue inhibitors of metalloproteinase-2 (TIMP-2) level in conditioned medium (collected in "Measurement of MMP-2 and MMP-9 activity by zymography" section) were detected with a TIMP-2 ELISA kit according the instruction.

Measurement of E-cadhedrin and,81-integrin expression in cell surface

Salidroside-treated cells ([10.sup.6]) were washed once in PBS sup-plemented with 0.5% FBS and 0.25% bovine serum albumin (BSA). Cells were then incubated for 30 min at 4[degrees]C in PBS with rabbit anti-human E-cadherin monoclonal antibody (FITC con-jugated) or rabbit anti-human 131-integrin monoclonal antibody (FITC conjugated). Subsequently, cells were washed twice and fixed in PBS until they were analyzed by flow cytometry (EPICSV, Coulter Instruments, Miami, FL).

Assay of PKC activation

Nonradioactive PKC assays were done with PepTag PKC assaykit (Promega, Madison, WI) according to manufacturer's instructions.Briefly, 20 [micro]g protein in particulate fraction of the cellular extract was incubated with PepTag PKC substrate and lipid activator at 30 [degrees]C for 30 min followed by inactivation at 95 [degrees]C for 5 min. After electrophoresis in 0.8% agarosegel for 15 min, the phosphorylated PKC and unphosphorylated PKC substrate migrating toward the anode and cathode, respectively, were detected under UV light.

Assay of ERK1/2 activation

HT1080 cells were lysed in buffer containing 50 mmol/1 Tris at pH 7.4, 50 mmo1/1 NaC1, 0.1% Triton X-100, 0.1% SDS, 0.3 mmo1/1 sodium orthovanadate, 50 mmo1/1 NaF, 1 mmo1/1 dithiotheritol, 10 mg/1 leupeptin, and 5 mg/l aprotinin. The protein concentrations of the resulting cell lysates were determined using a BCA protein assay kit (Pierce, Rockford, IL USA). An aliquot of each extract (40 [micro]g protein) was fractionated by electrophoresis in an SDS-polyacrylamide gel and transferred to a PVDF membrane. Membranes were blocked in 20 mmolil Tris (pH 7.6) and 250 mmol/l NaCl containing 5% drymilk and probed with antibodies against phosphorylated ERK 1/2 and ERK 1/2 (Santa Cruz Biotechnology, Santa Cruz, CA). For detection, enhanced chemiluminescence reagent (Pierce) was used with horseradish peroxidase-conjugated anti-mouse IgG or anti-goat IgG secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA). Reactive proteins were visualized using a chemiluminescence kit (Santa Cruz Biotechnology, Santa Cruz, CA) according to the manufacturer's instructions.

Statistical analysis

Data are presented as means S.E.M. from at least three independent experiments and evaluated by analysis of variance (ANOVA) followed by Student-Newman-Keuls test. Values of p < 0.05 were considered statistically significant.

Results

Salidroside slightly inhibited the proliferation of HT1080 cells

The effect of salidroside on HT1080 cells proliferation was determined by MIT assay after 24 h of exposure, cells viability was measured by trypan blue exclusion test. As shown in Fig. 2A, no obvious lethal effect was found after 24 h treatment with different concentrations salidroside (from 10 to 100 [micro]mo1/1), meanwhile salidroside just slightly inhibited the proliferation of HT1080 cells under 50 [micro]mo1/1. Compared to the control group, the 40 [micro]molfisalidroside-treated group showed no obvious morphological changes (Fig. 2B). Then the concentrations 10, 20 and 40 p/mo1/1 of salidroside were chose to study its anti-metastasis effect.

Salidroside inhibited the migration and invasion of HT1080 cells in vitro

In the in vitro migration assay, confluent monolayers of cells were wounded with a uniform scratch, rinsed to remove debris, and incubated in the absence or presence of salidroside (10, 20 and 40 [micro]mol/1) in serum free DMEM culture medium for 24 h. As shown in Fig. 3A, the wound closure level was 74.3% in the absence of salidroside, while after treatment with salidroside (10, 20 and 40 Rmol/l), the wound closure levels were significantly decreased (p<0.01 at 20 and 40 [micro]mol/l) in a concentration dependent way.

Invasion of HT1080 cells in vitro was measured by the Matrigel-coated transwell inserts. The result showed that salidroside treatment significantly inhibited the invasion of HT1080. After 24 h treatment with salidroside, the invaded cells were decreased to 58.6%, 41.1% and 18.2% (p < 0.01) at 10, 20 and 40 [micro]mo1/1 respectively (Fig. 3B).

Salidroside decreased the intracellular ROS formation

The fluorescent probe DCFH-DA was used to measure the effect of salidroside on the intracellular ROS level in HT1080 cells. As shown in Fig. 4, salidroside treatment significantly decreased the intracellular ROS level in HT1080 cells in a dose-dependent manner.

Salidroside decreased the MMP-2 activity and increased the TIMP-2 content in HT1080 cells conditioned medium

Since MMP-2 and MMP-9 play a pivotal role in tumor cell invasiveness, we wished to assess the effect of salidroside on MMP-2 and MMP-9 enzyme activities. MMP-2 and MMP-9 activities in conditioned media were detected by gelatin zymography assay. The result showed that salidroside treatment significantly suppressed the MMP-2 and MMP-9 activities in the conditioned medium of HT1080 cells (Fig. 5A). Whereas the endogenous MMPs inhibitor TIMP-2 content increased obviously in the conditioned medium (Fig. 56). These results indicated that a decrease in HT1080 cell invasion is a consequence, at least in part, of reduced activities of MMP-2/9 and increased TIMP-2 protein level.

Salidroside up-regulated the E-cadherin expression whereas down-regulated the [beta]l-integrin expression in HT1080 cells

To further confirm the ability of salidroside to inhibit metasta-sis of HT1080 cells, the expression of E-cadherin and [beta]1-integrin in HT1080 cells membrane were measured by flow cytometry with fluorescent-labeled antibodies. The results demonstrated that salidroside treatment induced the expression of E-cadherin in HT1080 cells. The positive rates of E-cadherin cells increased from 4.35% (control group) to 6.59%, 11.6% and 18.0% after 10, 20 and 40 [micro]mol/I salidroside treatment respectively (Fig. 6A). However, the same treatment with salidroside effectively down-regulated the [beta]1-integrin expression, which is an extracellular matrix adhesion receptor mediated the adhesion of cancer cells with normal tissue. The positive rates of [beta]1-integrin cells decreased from 35.4% (control group) to 15.9%, 9.37% and 4.58% after 10, 20 and 40 [micro]mol/l salidroside treatment respectively (Fig. 6B).

Salidroside inhibited activation of PKC

There are reports suggesting that PKC may also be activated by ROS to mediate many cellular effects (Gopalakrishna and Jaken 2000). Interestingly, ROS may amplify PKC signaling such as observed in highly glucose-treated human peritoneal mesothelial cells (Lee et al. 2004). In order to clarify whether salidroside may inactivate PKC to inhibit the PIKE-ERIC pathway by removing ROS. Thus, we used the PepTag PKC kit to assay the level of phosphorylated PKC. As shown in Fig. 7, the ratio of the intensity of phosphorylated PKC vs PKC for each sample represented the level of activated PKC. The ratio of the control samples was taken as 100%.Compared to the control group, salidroside-treated groups (10, 20 and 40 [micro]mol/l) were decreased to 54.7%, 36.3% and 26.7% (p <0.01 vs control). These results indicated that salidroside could inhibit the activation of PKC effectively.

Salidroside inhibited phosphorylation of ERK1/2

Since we have shown salidroside inhibited the cell metastasis and activities of MMP-2 and expression of the [beta]1-integrin, the underlying mechanisms were further investigated. Several studies have indicated the activatory ERK 1/2 are involved in activity of MMP-2, E-cadherin, and 131 -integrin on different cell types (Chen et al. 2005; Turner et al. 2007). To assess whether salidroside inhibits activation of ERK 1/2, we investigated the phosphorylated status of ERK 1/2 in HT1080 cells which were treated with various concentrations of salidroside for 24 h. Fig. 8 shows that salidro-side significantly inhibited the activation of ERK 1/2 as shown by decreasing the phosphorylation of ERK 1/2 and increasing the non-phosphorylated ERIC 1/2 in a dose-dependent manner.

Discussion

Salidroside has been reported to have many kinds of pharmacological effects in both in vivo and in vitro experimental models, but the effect of salidroside on cancer cells migration and invasion has not been reported. The present study demonstrated for the first time that salidroside treatment effectively inhibited the migration (Fig. 3A) and invasion (Fig. 3B) of HT1080 cells. Hu's works showed that salidroside induced apoptosis in cancer cells and the cytotoxic effects of salidroside were very strong (Hu et at 2010a,b), whereas there were other data suggest that salidroside has protective effects against apoptosis in cardiomyocytes and nerve cells and the cytotoxic effects were not obvious (Zhang et al. 2010; Thong et al. 2010). Wang's study proved that oral administration of 80, 40 and 20 mg/kg salidroside once daily for 7 days showed significantly antivirus effect in CVB3 virus infected mice (Wang et al. 2009). Furthermore, single intraperitoneal injection of salidroside (25, 50, 100, 200, 400, 800 mg/kg) had been proved to possess the sedative and hypnotic effect in mice (Li et al. 2007). No obvious toxicity was found in these studies, which demonstrated that salidroside was safe for usage at the concentration we studied. In our work, no obvious cytotoxic activity was found at 10-40 [micro]mol/l (Fig. 2), which indicated that the inhibitory effect of salidroside on HT1080 cells invasion and migration was not due to its cytotoxic effect. So we believed that salidroside possessed potent anti-metastasis activity with no serious toxicity.

Matrix metalloproteinases play a key role in tumor cells invasion, migration and tumor angiogenesis (Shimada et al. 2000; Rajoria et al. 2011; Yan et al. 2010). MMPs are the main family of proteolytic enzymes that facilitate tumor cell migration by degrading the basement membrane and other components of extra-cellular matrix (ECM). However, the activity of MMPs in extra cellular space is specifically inhibited by tissue inhibitors of metalloproteinases (TIMPs). TIMPs bind to the highly conserved zinc binding site of active MMPs at molar equivalence. Over expression of TIMP-2 can inhibit the activity of MMP-2 (Yan et al. 2010) and it also inhibits the invasive and metastatic behaviors of cancer cells. Our convincing evidence demonstrated that the salidroside could decrease MMP-2 and MMP-9 activities (Fig. 5A) and increase TIMP-2 expression (Fig. 5B) in HT1080 cells. Thus, salidroside may suppress the MMP-2 and MMP-9 activities, and increase the TIMP-2 expression to reduce the metastatic capabilities of HT1080 cells.

Invasion, generally leading to metastasis, is presented as the result of a balance between the activation of two sets of genes, coined i+ (invasion promotor) and i- (invasion suppressor) genes (Makrilia et al. 2009; Mareel et al. 1992). Experiments in vitro as well as in vivo have indicated that the homotypic homophilic epithelial cell-cell adhesion molecule E-cadherin is an i-gene product, it acts as an invasion suppressor (Khamis et al. 2011; Debruyne et al. 1999; Mareel et al. 1995; Mareel et al. 1994).But as know that E-cadherin is down-regulated during metastasis (Saegusa et al. 2000; Fogel et al. 2004; Vestweber and Blanks 1999; Flugy et al. 2002), thereby promoting cell invasion. The integrin family of heterogeneous cell-cell adhesion molecules are heterodimeric receptors involved in cell-to-cell and cell-to-extracellular matrix interactions (Albelda and Buck 1990).The integrin is an i+ gene product, it acts as an invasion promotor. A relationship has been found for several cancers, between metastatic potential of tumor cells during invasion through the basal membranes, and qualitative as well as quantitative changes in the integrin expression (Ramirez et al. 2011; Gehlsen et a1.1992; Prifti et al. 2008). Interestingly, we found that salidroside could not only increase the positive cells of E-cadherin (Fig. 6A), up-regulate the expression of E-cadherin, but also decrease the positive cells of [beta]1-integrin (Fig. 6B), down-regulate the expression of [beta]1-integrin.The results further confirmed the ability of salidroside to inhibit metastasis of HT1080 cells.

The MAPK sighaling cascade, including ERK 1/2, c-Jun N-terminal kinase (JNK), and P38, has also been implicated in the migration of numerous cell types (Huang et al. 2004). Specific inhibitors for ERIC may inhibit the migration of cells in response to extracellular matrix and growth factors, such as fibroblast growth factor and epidermal growth factor (Webb et al. 2000; Cheresh et al.1999; Cho and Klemke 2000). Moreover, inhibition of ERK by an antisense strategy was shown to prevent cell migration (Lai et al.2001). ROS production has been shown to be coupled with the sustained activation of the ERK signaling pathway for a variety of cellular effects, including apoptosis (Lee et al. 2005) and phagocytosis. Furthermore, a report showed that oxidative stress may induce invasive potential of mammary epithelial cells (Mori et al.2004). In addition, activations of PKC and ERK were shown to be responsible for oleic acid- and angiotensin II-induced vascular smooth muscle cell migration (Greene et al. 2001). Wu's work proved that ROS play a central role in mediating TPA-triggered sustained PKC and ERK signaling for regulation of gene expression of MMPs, integrins and E-cadherin that are responsible for EMT and migration of HepG2 (Wu et al. 2006). Our present work showed that salidroside treatment decreased the phosphorylation of ERK1 /2 in HT1080 cells (Fig. 8). Panossian et al. found that salidroside could reduce the phosphorylation of JNK, one of the MAPK family, to fight against fatigue and stress in rabbits (Panossian et al. 2007). The results obtained in vitro and in vivo indicated that salidroside could attenuate the activation of MAPK family pathway. However, the underlying mechanism was unclear. Our work showed that salidroside treatment decreased the intracellular ROS level in HT1080 cells significantly (Fig. 4) and inhibited the activation of PKC (Fig. 7), these might be associated with the inhibitory effect of salidroside on MAPK activation.

It is well known that inhibition of metastasis of cancer cells have important preventive and therapeutic benefits on cancer. The present study proved the anti-metastasis efficacy of salidroside and our data together with the results published previously suggested that the mechanisms might he due to its downregulation of the ROS-PKC-ERK1/2 signaling pathway. Therefore, we suggest that salidroside may be used as an effective constituent for anti-metastases products. Further studies are necessary to assess the anti-metastases capabilities of salidroside in vivo, especially, to demonstrate the effect on tumor cells prior to their escape from the primary tumor to form metastases. These further studies are now in progress.

Acknowledgements

This study was supported by the National Basic Research Program of China (973 Program) (2010CB834202) to H.Z., National Natural Science Foundation of China (No. 30760058) to LW., High Level Talent Start Fund Project of Shihezi University (No.RCZX200667) to Z.W., Basic Research Project of Xinjiang Production and Construction Corps (No.2007JCO3) to Z.W., and Special Grade of China Postdoctoral Science Foundation (No. 200902313) to Z.W.

* Corresponding author at: Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, PR China. Tel.: +86 931 4969747; fax: +86 931 4969200.

** Corresponding author at: Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, PR China. Tel.: +86 931 4969344.

E-mail addresses: wangzh@impcas.ac.cn (Z. Wang), zhangh@impcas.ac.cn (H. Zhang).

References

Albelda, S.M., Buck, CA., 1990. Integrins and other cell adhesion molecules. FASEB J. 4, 2868-2880.

Bogenrieder, T., Herlyn, M., 2003. Axis of evil: molecular mechanisms of cancer metastasis. Oncogene 22, 6524-6536.

Cao, L.L., Du, G.H., Wang, M.W., 2005. Effect of salidroside on mitochondria injury induced by sodium azide. Yao Xue Xue Bao 40, 700-704.

Chen, P.N., Hsieh, Y.S., Chiou, H.L., Chu, S.C., 2005. Silibinin inhibits cell invasion through inactivation of both 113K-Akt and MAPK signaling pathways. Chem. Biol. Interact. 156, 141-150.

Cheresh, D.A., Leng, J., Klemke, R.L., 1999. Regulation of cell contraction and mem-brane ruffling by distinct signals in migratory cells. J. Cell Biol. 146, 1107-1116. Chiarugi, P., 2005. PTPs versus PTKs: the redox side of the coin. Free Radic. Res. 39, 353-364.

Cho, S.Y., Klemke, R.L., 2000. Extracellular-regulated kinase activation and CAS/Crk coupling regulate cell migration and suppress apoptosis during invasion of the extracellular matrix. J. Cell Biol. 149, 223-236.

Debruyne, P., Vermeulen, S., Mareel, M., 1999. The role of the E-cadherin/catenin complex in gastrointestinal cancer. Acta Gastroenterol. Belg. 62, 393-402.

Dutta, A., Sen, T., Banerji, A., Das, S., Chatterjee, A., 2009. Studies on multifunctional effect of all-trans retinoic acid (ATRA) on matrix metalloproteinase-2 (MMP2) and its regulatory molecules in human breast cancer cells (MCF-7). J. Oncol. [Epub].

Flugy, A.M., D'Amato, M., Russo, D., Di Bella, M.A., Alaimo, G., Kohn, E.C., De Leo, G., Alessandro, R., 2002. E-selectin modulates the malignant properties of T84 colon carcinoma cells. Biochem. Biophys. Res. Commun. 293, 1099-1106.

Fogel, M., Huszar, M., Altevogt, P., Ben-Arie, A., 2004. Ll (CD171) as a novel biomarker for ovarian and endometrial carcinomas. Expert Rev. Mol. Diagn. 4, 455-462.

Forman, KJ., Torres, M., Fukuto, J., 2002. Redox signaling. Mol. Cell Biochem. 234, 49-62.

Gehlsen, K.R., Davis, G.E., Sriramarao, P., 1992. Integrin expression in human melanoma cells with differing invasive and metastatic properties. Clin. Exp. Metastasis 10, 111-120.

Goodison, S., Kawai, K., Hihara, J., Jiang, P., Yang, M., Urquidi, V., Hoffman, R.M., Tartu, D., 2003. Prolonged dormancy and site-specific growth potential of cancer cells spontaneously disseminated from nonmetastatic breast tumors as revealed by labeling with green fluorescent protein. Clin. Cancer Res. 9, 3808-3814.

Gopalakrishna, R., Jaken, S., 2000. Protein kinase C signaling and oxidative stress. Free Radic. Biol. Med. 28, 1349-1361.

Greene, Et., Lu, G., Zhang, D., Egan, B.M., 2001. Signaling events mediating the additive effects of oleic acid and angiotensin II on vascular smooth muscle cell migration. Hypertension 37, 308-312.

Gupta, G.P., Massague, J., 2006. Cancer metastasis: building a framework. Cell 127, 679-695.

Hanahan, D., Weinberg, R.A., 2000. The hallmarks of cancer. Cell 100, 57-70. Huang, C., Jacobson. K., Schaller, M.D., 2004. MAP kinases and cell migration. J. Cell Sci. 117, 4619-4628.

Hu, X., Zhang, X., Qiu, S., Yu, D., Lin, S., 2010a. Salidroside induces cell-cycle arrest and apoptosis in human breast cancer cells. Biochem. Biophys. Res. Commun. 398, 62-67.

Hu, X., Lin, S., Yu, D., Qiu, S., Zhang, X., Mei, R., 2010b. A preliminary study: the antiproliferation effect of salidroside on different human cancer cell lines. Cell Biol. Toxicol. 26, 499-507.

Kanupriya, Prasad, D., Sai Ram, M., Kumar, R., Sawhney, R.C., Sharma, S.K., Ilavazhagan G., Kumar, D., Banerjee, P.K., 2005. Cytoprotective and antioxidant activity of Rhodiola imbricata against tert-butyl hyd roperoxide induced oxidative injury in 1..1-937 human macrophages. Mol. Cell. Biochem. 275, 1-6.

Khamis, Z.I., Iczkowski, K.A., Sang, Q.X., 2011. Metastasis suppressors in human benign prostate, intraepithelial neoplasia, and invasive cancer: their prospects as therapeutic agents. Med. Res. Rev. 16 [Epub ahead of print].

Kucinskaite, A., Briedis, V., Savickas, A., 2004. Experimental analysis of therapeutic properties of Rhodiofa roses L and its possible application in medicine. Medicina (Kaunas) 40, 614-619.

Lai, C.F., Chaudhary, L., Fausto, A., Halstead, L.R., Ory, D.S., Avioli, L.V., Cheng, S.L., 2001. Erk is essential for growth, differentiation, integrin expression, and cell function in human osteoblastic cells. J. Biol. Chem. 276, 14443-14450.

Lee, E.R., Rang, YJ., Kim, J.H., Lee, H.T., 2005. Modulation of apoptosis in HaCaT keratinocytes via differential regulation of ERIC signaling pathway by flavonoids. J. Biol. Chem. 280, 31498-31507.

Lee, H.B., Yu, M.R., Song, J.S., Ha, H., 2004. Reactive oxygen species amplify protein kinase C signaling in high glucose-induced fibronectin expression by human peritoneal mesothelial cells. Kidney Int. 65, 1170-1179.

Lee, IC., Esselman, W.J., 2002. Inhibition of PTPs by 1-1202 regulates the activation of distinct MAPIC pathways. Free Radic. Biol. Med. 33, 1121-1132.

Li, T., Xu, G., Wu, L., Sun, C., 2007. Pharmacological studies on the sedative and hypnotic effect of salidroside from the Chinese medicinal plant Rhodiofa sachalinensis. Phytomedicine 14, 601-604.

Li, D., Wang, Z., Chen, H., Wang, J., Meng, Q., Shang, J., Li, J., 2009. Isoliquiritigenin induces monocytic differentiation of HL-60 cells. Free Radic. Biol. Med. 46, 731-736.

Lin, S.J., Shyue, S.K., Liu, P.L., Chen, Y.H., Ku, H.H., Chen, j.W.,Tam, K.B., Chen, Y.L., 2004. Adenovirus-mediated overexpression of catalase attenuates oxLDL-induced apoptosis in human aortic endothelial cells via AP-1 and C-Jun N-terminal kinase/extracellular signal-regulated kinase mitogen-activated protein kinase pathways. J. Mol. Cell. Cardiol. 6, 129-139.

Lo, 1.C., Shih, J.M., Jiang, M.j., 2005. Reactive oxygen species and ERK 1/2 mediate monocyte chemotactic protein-1-stimulated smooth muscle cell migration. J. Biomed. Sci. 12, 377-388.

Mao, G.X., Deng, H.B., Yuan, L.G., Li, D.D., Li, Y.Y., Wang, Z., 2010. Protective role of salidroside against aging in a mouse model induced by D-galactose. Biomed. Environ. Sci. 23, 161-166.

Makrilia, N., Kollias, A.. Manolopoulos, L, Syrigos, K., 2009. Cell adhesion molecules: role and clinical significance in cancer. Cancer Invest. 27, 1023-1037.

Mareel, M., Vleminckx, K., Vermeulen, S., Bracke, M., Van Roy. F., 1992. E-cadherin expression: a counterbalance for cancer cell invasion. Bull. Cancer 79, 347-355.

Mareel, M., Vleminckx, K., Vermeulen, S., Yan, G., Bracke, M., Van Roy, F., 1994. Downregulation in vivo of the invasion-suppressor molecule E-cadherin in experimental and clinical cancer. Princess Takamatsu Symp. 24, 63-80.

Mareel, M., Bracke, M., Van Roy, F., 1995. Cancer metastasis: negative regulation by an invasion-suppressor complex. Cancer Detect. Prey. 19, 451-464.

Martinez, V.G., Williams. KJ., Stratford, I.J., Clynes, M., O'Connor, R., 2008. Over-expression of cytochrome P450 NADPH reductase sensitises MDA 231 breast carcinoma cells to 5-fluorouracil: possible mechanisms involved. Toxicology 22, 582-588.

Mosmann, T., 1983. Rapid colon metric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65, 55-63.

Mori, K., Shibanuma, M., Nose, IC, 2004. Invasive potential induced under longterm oxidative stress in mammary epithelial cells. Cancer Res. 64, 7464-7472.

Nishikawa, M., 2008. Reactive oxygen species in tumor metastasis. Cancer Lett. 266, 53-59.

Panossian, A., Hambartsumyan, M., Hovanissian, A., Gabrielyan, E., Wikman, G., 2007. The adaptogens rhodiola and schizandra modify the response to immobilization stress in rabbits by suppressing the increase of phosphorylated stress-activated protein kinase, nitric oxide and cortisol. Drug Targets Insights 1, 39-54.

Prifti, S., Zourab, Y., Koumouridis, A., Bohlmann, M., Strowitzki, T., Rabe, T., 2008. Role of integrins in invasion of endometrial cancer cell lines. Gynecol. Oncol. 84. 12-20.

Radisky, D.C., Levy, D.D., Littlepage, LE., Liu, H., Nielson, C.M., Fata, J.E., Leake, D., Godden, E.L., Albertson, D.G., Nieto, M.A., Werb, Z., Bissell, M.J., 2005. Racl b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability. Nature 436, 123-127.

Rajoria, S., Suriano, R., George, A., Shanmugam, A., Schantz, SP., Geliebter, J., Tiwari, R.K., 2011. Estrogen induced metastatic modulators MMP-2 and MMP-9 are targets of 3,3'-diindolylmethane in thyroid cancer. PLoS One 18, e15879.

Ramirez, N.E., Zhang, Z., Madamanchi, A., Boyd, K.L., O'Rear, L.D., Nashabi, A., Li, Z., Dupont, W.D., Zijlstra, A., Zutter, M.M., 2011. The [alpha]2[beta]1 integrin is a metastasis suppressor in mouse models and human cancer. J. Clin. Invest. 121, 226-237.

Reth, M., 2002. Hydrogen peroxide as second messenger in lymphocyte activation. Nat. Immunol. 3, 1129-1134.

Rhyu, D.Y., Yang, Y., Ha, FL, Lee, G.T., Song, J.S., Uh, S.T., Lee, H.B., 2005. Role of reactive oxygen species in TGF-betal-induced mitogen-activated protein kinase activation and epithelial-mesenchymal transition in renal tubular epithelial cells. J. Am. Soc. Nephrol. 16, 667-675.

Saegusa, M., Machida, D., Okayasu, 1., 2000. Loss of DCC gene expression during ovarian tumorigenesis: relation to tumour differentiation and progression. Br.). Cancer 82, 571-578.

Sauer, H., Wartenberg, M., Hescheler, J., 2001. Reactive oxygen species as intracellular messengers during cell growth and differentiation. Cell. Physiol. Biochem. 11, 173-186.

Shackelford, R.E., Kaufmann, W.K., Paules, R.S., 2000. Oxidative stress and cell cycle checkpoint function. Free Radic. Biol. Med. 28, 1387-1404.

Shimada, T., Nakamura, H., Yamashita, K., Kawata, R., Murakami, Y., Fujimoto, N., Sato, H., Seiki, M., Okada, Y., 2000. Enhanced production and activation of

progelatinase A mediated by membrane-type 1 matrix metalloproteinase in human oral squamous cell carcinomas: implications for lymph node metastasis. Clin. Exp. Metastasis 18, 179-188.

Storz, P., 2005. Reactive oxygen species in tumor progression. Front. Biosci. 10, 1881-1896.

Turner, NA, Aley, P.K., Hall, K.T., Warburton, P., Galloway, S., Midgley, L, O'Regan, D.J., Wood, 1.C., Ball, S.G., Porter, ICE., 2007. Simvastatin inhibits TNFalphainduced invasion of human cardiac myofibroblasts via both MMP-9-dependent and - independent mechanisms. J. Mol. Cell. Cardiol. 43, 168-176.

Vesrweber, D, Blanks, J.E., 1999. Mechanisms that regulate the function of the selectins and their ligands. Physiol. Rev. 79, 181-213.

Wang, H., Ding, Y., Zhou, J., Sun, X., Wang, S., 2009. The in vitro and in vivo antiviral effects of salidroside from Rhodiola rosea L. against coxsackievirus B3. Phytomedicine 16, 146-155.

Webb, D.J., Nguyen, D.H., Gonias, S.L., 2000. Extracellular signal-regulated kinase functions in the urokinase receptor-dependent pathway by which neutralization of low density lipoprotein receptor-related protein promotes fibrosarcoma cell migration and Matrigel invasion. J. Cell Sci. 113, 123-134.

Weis, M., Heeschen, C., Glassford, A.J., Cooke, J.P., 2002. Statins have biphasic effects on angiogenesis. Circulation 105, 739-745.

Wu, H.M., Zhu, S.L., He, L.J., Liu, Y.H., Xie, D., 2009. Clinical significance of macrophage migration inhibitory factor in invasion of ovarian cancer. Chin. J. Cancer 28, 1054-1060.

Wu, W.S., 2006. The signaling mechanism of ROS in tumor progression. Cancer Metastasis Rev. 25, 695-705.

Wu, W.S., Tsai, R.K., Chang, C.H., Wang, S., Wu, J.R., Chang, Y.X., 2006. Reactive oxygen species mediated sustained activation of protein kinase C alpha and extracellular signal-regulated kinase for migration of human hepatoma cell HepG2. Mol. Cancer Res. 4, 747-758.

Wu, W.S., Wu, J.R., Hu, CT., 2008. Signal cross talks for sustained MAPK activation and cell migration: the potential role of reactive oxygen species. Cancer Metastasis Rev. 27, 303-314.

Yan, L, Lin, B., Gao, L, Gao, S., Liu, C., Wang, C., Wang, Y., Zhang, S., Iwamori, M., 2010. Lewis (y) antigen overexpression increases the expression of MMP-2 and MMP-9 and invasion of human ovarian cancer cells. Int. J. Mol. Sci. 11, 4441-4452.

Zhong, H., Xin, H., Wu, LX., Zhu, Y2., 2010. Salidroside attenuates apoptosis in ischemic cardiomyocytes: a mechanism through a mitochondria-dependent pathway. J. Pharmacol. Sci. 114, 399-408.

Zhang, L, Yu, H., Sun, Y., Lin, X., Chen, B., Tan, C., Cao, G., Wang, Z., 2007. Protective effects of salidroside on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells. Eur. J. Pharmacol. 564, 18-25.

Zhang, L., Yu, H., Zhao, X., Lin, X., Tan, C., Cao, G., Wang, Z., 2010. Neuroprotective effects of salidroside against beta-amyloid-induced oxidative stress in SH-SYSY human neuroblastoma cells. Neurochem. Mt. 57, 547-555.

Chao Sun (a), (b), (c), (d), Zhenhua Wang(a), (b), (c), (e), * Qiusheng Zheng(e) Hong Zhang (a), (b), (c),**

(a.) Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, PR China

(b.) Key Laboratory of Heavy Ion Radiation Medicine of Chinese Academy of Sciences, Lanzhou, PR China

(c.) Key Laboratory of Heavy Ion Radiation Medicine of Gansu Province, Lanzhou, PR China

(d.) Graduate University of Chinese Academy of Sciences, Beijing, PR China

(e.) Key Laboratory of Xinjiang Endemic Phytomedicine Resources, Ministry of Education, School of Pharmacy, Shihezi University, Shihezi, PR China

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doi:10.1016/j.phymed.2011.09.070
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Author:Sun, Chao; Wang, Zhenhua; Zheng, Qiusheng; Zhang, Hong
Publication:Phytomedicine: International Journal of Phytotherapy & Phytopharmacology
Article Type:Report
Geographic Code:9CHIN
Date:Mar 1, 2012
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