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Detection of PRL-3 protein in the preoperative serum of patients with colorectal cancer.

INTRODUCTION

Numerous research on carcinogenesis in colorectal cancer have been presented PRL-3 protein as one of the major poor prognostic marker for patients but also as a promising target for the cancer treatment. PRL-3 protein (phosphatase of regenerating liver-3) belongs to the family of protein tyrosine phosphatases, which play an important role in regulating physiological and pathological cell processes. Specifically, it is involved in reconstructing of the cytoskeleton, regulating adhesion and cell cycle of the cancer cells, and epithelial-mesenchymal transition. Through these mechanisms PRL-3 protein participates in invasion, migration, metastasis and angiogenesis. Studies indicate the PRL-3 protein takes part especially in metastases of colon cancer [1].

The family of PRL phosphatases consist of three proteins (PRL-1, PRL-2, PRL-3), all of which have a unique C-terminal prenylation motif. They show 75% homology in amino acid sequence and have a molecular weight of about 20kDa [2]. The gene responsible for encoding PRL-3 protein is located at chromosome 8q24.3 [3]. Human PRL-3 molecule is composed of 173 amino acids and is a monomer with a complex structure. Enzyme active side is marked by a signature sequence HCXXGXXR where Cys104 is the enzymatic nucleophile and Arg110 coordinates with the phosphate group on phosohotyrosine [4]. In contrast to other phosphatases, catalytic core of PRL-3 does not contain serine or threonine, important for the catalytic activity of the other phosphatases [5]. PRL-3 molecule contains C-terminal consensus sequence for prenylation (CAAX motif) which is responsible for farnesylation of the protein. As a result, it can be found in the membranes and intracellular structures when is prenylated and in the nucleus as the non-prenylated form [6-8]. In healthy subjects, PRL-3 protein is detected in cardiomyocytes, skeletal muscles and to a certain degree in the pancreas [2, 3]. In colorectal cancer, PRL-3 protein is found in a small percentage of primary tumors and in almost all metastases [1]. The cellular localization of PRL3 protein was closely associated with its function in tumor metastases [9].

The aim of study was to detect PRL-3 protein in sera of patients with colorectal cancer.

MATERIALS AND METHODS

Patients

The study group consisted of 22 patients diagnosed with colorectal cancer (13 men and 9 women) treated surgically in the 2nd Department of General and Gastroenterological Surgery in the Medical University of Bialystok in 2007-2009. The pathological diagnosis confirmed colorectal cancer and classified as adenocarcinoma type without mucin in 20/22 of patients and adenocarcinoma with mucous component in 2/22 of patients. The investigated tumors were classified as moderately (20/22 of patients) and poorly differentiated (2/22 of patients). According to TNM classification, tumor infiltrated a submucosa (pT1) in 1 case, muscle layer (pT2) in 2 cases and subserosa (pT3) in 19 cases. The metastases to local lymph nodes were observed in 9/22 of cases and the presence of metastases to distant organs was noted in 7/22 of cases.

Study material consisted of serum samples obtained from both the blood of the patients with colorectal carcinoma collected prior to the surgery.

Blood serum was stored at -800C immediately after centrifugation until the assay was performed.

The control group consisted of 10 healthy volunteers (5 males and 5 females aged 49-80).

The study received the approval of the local Bioethics Committee. All the participants received information about method and purpose of study and signed consent forms prior to the examination.

ELISA method

To determine the concentration of PRL-3 protein in serum it was used commercially available ELISA kit (Uscn Life Science Inc.) according to the manufacturer's instructions. The sera were slowly defrosted to room temperature. The detectable concentration of PRL-3 protein range from 0.312-20ng/mL.

Statistics

Statistical analysis was conducted based on the STATISTICA 8.0 program. In order to compare the two groups, the U Mann-Whitney test was used. Correlations between the protein level in serum and clinical-pathological parameters were calculated by the Spearman's correlation coefficient tests. The level of significance was <0.05.

RESULTS

Serum PRL-3 levels in colorectal cancer and control patients

In the control group, 4 cases had the levels of PRL-3 protein below minimum value indicated by kit (<0.115ng/ml) and those results were defined as a level 0 (absent protein) whereas the concentration of PRL-3 protein ranged from 0.263-1.158ng/ml) in 6 cases.

In the study group with colorectal cancer, the level of protein in preoperative sera was negative (<0.115ng/ml) in 7 cases while the mean value of PRL-3 concentration was 0.616ng/ml (range 0.206-2.072ng/ml) in 15 patients. No statistically significant differences between the study group and healthy controls were observed (Tab. 1).

Correlation between serum PRL-3 levels and clinicopathological parameters in colorectal cancer patients

No correlations of preoperative serum PRL-3 levels in colorectal cancer patients with age, gender, tumor localization, histological type, stage, local lymph node involvement, distant organs metastases and Duke's classification were observed. However, our data showed that increase in level of PRL-3 protein in sera of patients with colorectal cancer is associated with greater tumor size (p<0.05) (Tab.2).

DISCUSSION

Since it has been demonstrated that the PRL-3 protein is significantly involved in the development of cancer, it began to see it as a possible target in anti-cancer treatment. It was also observed that PRL-3 knockdown inhibits the tumor growth through reducing the potential for its expansion and the activation of apoptosis. It reduces the lymph node involvement too [10].

Therefore, it has been started to explore the inhibitors of PRL-3 protein such as TGFp, PCBP1 [11], thienopryridone [12], curcumin [13], methanolic extract of the roots of Rubia Akane [14] as possible attractive targets for cancer therapy. However, antibody therapy gives the best hope for cancer patients. Guo et al. [9] presented the treatment of a mouse model of chimeric antibodies which blocked the intracellular PRL-3 protein. They observed that those antibodies specifically reduce the metastases formation related to the expression of PRL-3 protein. Since the research on therapy associated with PRL-3 protein has been improved, an attempt was made to select the patients too. The mechanism of this protein overexpression is still not understood so Ooki et al. [15] evaluated the PRL-3 gene amplification in patients with gastric cancer based on the assessment of HER2 gene status. They demonstrated that gene amplification occurred in 20% of cases with the positive expression of PRL-3 protein in tumour tissue. Moreover, they stated that the assessment of the presence of PRL-3 expression, but not the level of expression, may have a significant potential for the classification of PRL-3-linked therapy. A similar research should be carried out on the group of patients with colorectal cancer.

In our study, we tried to detect the extracellular presence of PRL-3 protein in serum. While it has been proved that PRL-3 protein presents in sera of patients with colorectal cancer, we did not observe a significant differences between the levels of this protein compare to sera of healthy patients. However, the question is if the presence of PRL-3 in serum may influence on antibody therapy.

The level of PRL-3 protein was not found to correlate with clinicopathological parameters, except tumor size. In cases of tumor greater than 5 cm, an increase in the level of PRL-3 protein was observed. Nevertheless, we cannot establish the source of PRL-3 protein.

CONCLUSIONS

1. PRL-3 protein was fund to be present in sera of patients with colorectal cancer.

2. However our study indicates lack of clinical usefulness of determining the levels of PRL-3 in the sera of patients with colorectal cancer.

Conflicts of interest

The authors declare no conflict of interest.

REFERENCES

[1.] Guzinska-Ustymowicz K, Pryczynicz A. PRL-3, an emerging marker of carcinogenesis, is strongly associated with poor prognosis. Anticancer Agents Med Chem. 2011 Jan; 11(1):99-108. Review.

[2.] Zeng Q, Hong W, Tan YH. Mouse PRL-2 and PRL-3, two potentially prenylated protein tyrosine phosphatases homologous to PRL-1. Biochem Biophys Res Commun. 1998 Mar 17; 244(2):421-7.

[3.] Matter WF, Estridge T, Zhang C, Belagaje R, Stancato L, Dixon J, Johnson B, Bloem L, Pickard T, Donaghue M, Acton S, Jeyaseelan R, Kadambi V, Vlahos CJ. Role of PRL-3, a human muscle-specific tyrosine phosphatase, in angiotensin-II signaling. Biochem Biophys Res Commun. 2001 May 25; 283(5):1061-8.

[4.] Kim KA, Song JS, Jee J, Sheen MR, Lee C, Lee TG, Ro S, Cho JM, Lee W, Yamazaki T, Jeon YH, Cheong C. Structure of human PRL-3, the phosphatase associated with cancer metastasis. FEBS Lett. 2004 May 7; 565(1-3):181-7.

[5.] Kozlov G, Cheng J, Ziomek E, Banville D, Gehring K, Ekiel I. Structural insights into molecular function of the metastasis-associated phosphatase PRL-3. J Biol Chem. 2004 Mar 19; 279(12):11882-9.

[6.] Guo K, Li J, Tang JP, Koh V, Gan BQ, Zeng Q. Catalytic domain of PRL-3 plays an essential role in tumor metastasis: formation of PRL-3 tumors inside the blood vessels. Cancer Biol Ther. 2004 Oct; 3(10):945-51.

[7.] Zeng Q, Si X, Horstmann H, Xu Y, Hong W, Pallen CJ. Prenylation-dependent association of protein-tyrosine phosphatases PRL-1,-2, and-3 with the plasma membrane and the early endosome. J Biol Chem. 2000 Jul 14; 275(28):21444-52.

[8.] Bessette DC, Qiu D, Pallen CJ. PRL PTPs: mediators and markers of cancer progression. Cancer Metastasis Rev. 2008 Jun; 27(2):231-52.

[9.] Guo K, Tang JP, Jie L, Al-Aidaroos AQ, Hong CW, Tan CP, Park JE, Varghese L, Feng Z, Zhou J, Chng WJ, Zeng Q. Engineering the first chimeric antibody in targeting intracellular PRL-3 oncoprotein for cancer therapy in mice. Oncotarget. 2012 Feb; 3(2):158-71.

[10.] Matsukawa Y, Semba S, Kato H, Koma Y, Yanagihara K, Yokozaki H. Constitutive suppression of PRL-3 inhibits invasion and proliferation of gastric cancer cell in vitro and in vivo. Pathobiology. 2010; 77(3):155-62.

[11.] Wang H, Vardy LA, Tan CP, Loo JM, Guo K, Li J, Lim SG, Zhou J, Chng WJ, Ng SB, Li HX, Zeng Q. PCBP1 suppresses the translation of metastasis-associated PRL-3 phosphatase. Cancer Cell. 2010 Jul 13; 18(1):52-62.

[12.] Daouti S, Li WH, Qian H, Huang KS, Holmgren J, Levin W, Reik L, McGady DL, Gillespie P, Perrotta A, Bian H, Reidhaar-Olson JF, Bliss SA, Olivier AR, Sergi JA, Fry D, Danho W, Ritland S, Fotouhi N, Heimbrook D, Niu H. A selective phosphatase of regenerating liver phosphatase inhibitor suppresses tumor cell anchorage-independent growth by a novel mechanism involving p130Cas cleavage. Cancer Res. 2008 Feb 15; 68(4):1162-9.

[13.] Wang L, Shen Y, Song R, Sun Y, Xu J, Xu Q. An anticancer effect of curcumin mediated by down-regulating phosphatase of regenerating liver-3 expression on highly metastatic melanoma cells. Mol Pharmacol. 2009 Dec; 76(6):1238-45.

[14.] Moon MK, Han YM, Lee YJ, Lee LH, Yang JH, Kwon BM, Kim DK. Inhibitory activities of anthraquinones from Rubia akane on hosphatase egenerating liver-3. Arch Pharm Res. 2010 Nov; 33(11):1747-51.

[15.] Ooki A, Yamashita K, Kikuchi S, Sakuramoto S, Katada N, Waraya M, Kawamata H, Nishimiya H, Nakamura K, Watanabe M. Therapeutic potential of PRL-3 targeting and clinical significance of PRL-3 genomic amplification in gastric cancer. BMC Cancer. 2011 Apr 6; 11:122.

Pryczynicz A. (1) *, Dymicka-Piekarska V. (2), Niewiarowska K1, Gryko M. (3), Niksa M. (1), Hawryluk M. (1), Kemona A. (1), Famulski W. (4)

(1) Department of General Pathomorphology, Medical University of Bialystok, Poland

(2) Department of Clinical Laboratory Diagnostics, Medical University of Bialystok, Poland

(3) 2nd Department of General and Gastroenterological Surgery, Medical University of Bialystok, Poland

(4) Department of Medical Pathomorphology, Medical University of Bialystok, Poland

Corresponding author:

Anna Pryczynicz

Department of General Pathomorphology

Medical University of Bialystok

ul. Waszyngtona 13, 15-269 Bialystok, Poland

Tel/Fax: +48 85 7485996

e-mail: pryczynicz.anna@gmail.com

Received: 10.05.2013

Accepted: 21.05.2013
Table 1. Serum PRL-3 levels in patients with
colorectal cancer and control group.

                        PRL-3 (ng/ml)

         N    Mean    Median    SD     Range    p value

Normal   10   0.251   0.036    0.383   0-.158     NS
Tumor    22   0.616   0.500    0.637   0-.072

Mann-Whitney U-test. NS-non significant.

Table 2. Correlations between serum PRL-3 levels and
clinicopathological parameters in colorectal cancer patients.

                                               PRL-3 (ng/ml)
                                                                  p
Parameter                            N    Mean    Coefficient   value

Age                     <60          9    0.581     -0.099       NS
                        >60          13   0.640

Gender                  Male         13   0.501     -0.772       NS
                       Female        9    0.782

Localization           Colon         14   0.622      0.067       NS
                       Rectum        8    0.606

Adenocarcinoma      Nonmucinous      20   0.646     -0.570       NS
  type                Mucinous       2    0.312

Grade of               Poorly        20   0.616      0.455       NS
  malignancies     differentiated
                     Moderately      2    0.615
                   differentiated

pT stage                 1           1    0.891     -0.759       NS
                         2           2    0.539
                         3           19   0.610

Lymph node             Absent        13   0.466      0.983       NS
  metastasis          Present        9    0.833

Distant            Absent Present    15   0.642     -0.385       NS
  metastasis                         7    0.56

Duke stage               A           2    0.862      0.019       NS
                         B           7    0.354
                         C           3    0.879
                         D           10   0.671

Tumor size              <5cm         12   0.421      0.511      <0.05
                  [greater than or   10   0.717
                    equal to] 5cm

Spearman's correlation coefficient test. NS-non significant
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Article Details
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Author:Pryczynicz, A.; Dymicka-Piekarska, V.; Niewiarowska, K.; Gryko, M.; Niksa, M.; Hawryluk, M.; Kemona,
Publication:Progress in Health Sciences
Article Type:Clinical report
Geographic Code:4EXPO
Date:Jun 1, 2013
Words:2182
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