Silencing of pancreatic adenocarcinoma upregulated factor by RNA interference inhibits the malignant phenotypes of human colorectal cancer cells

Liu,Peng-Fei; Wu,Yun-Yun; Hu,You; Wang,Lei; He,Song-Bing; Zhu,Yi-Bei; Zhu,Xin-Guo

doi:10.3892/or.2013.2478

Silencing of pancreatic adenocarcinoma upregulated factor by RNA interference inhibits the malignant phenotypes of human colorectal cancer cells

Authors:
- Peng-Fei Liu
- Yun-Yun Wu
- You Hu
- Lei Wang
- Song-Bing He
- Yi-Bei Zhu
- Xin-Guo Zhu
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Affiliations: Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China, Department of Immunology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China
Published online on: May 15, 2013 https://doi.org/10.3892/or.2013.2478
Pages: 213-220

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Abstract

Aberrant expression of pancreatic adenocarcinoma upregulated factor (PAUF), a novel secretory protein, has been reported in several types of cancer. However, in colorectal cancer (CRC), whether PAUF also plays its oncogenic role through the Wnt/β-catenin pathway and its effect in regulating malignant phenotypes of CRC is unknown. In this study, we detected PAUF and β-catenin expression levels by immunohistochemical analysis and real-time PCR in CRC tissues, adjacent non-tumor tissues (NATs) and 5 CRC cell lines. The results demonstrated that the expression of PAUF and β-catenin in tumor tissues was higher than in NATs. Moreover, the expression of PAUF was correlated with the expression of β-catenin in both tumor tissues and NATs. The HCT116 cell line, which has the highest PAUF expression of the 5 cell lines, was transfected with small interfering RNA (siRNA) targeting on PAUF, which significantly downregulated the expression of PAUF in cancer cells. Successful transfection was confirmed by using RT-PCR and western blot analysis. Further studies demonstrated that PAUF-siRNA inhibited the proliferation of CRC cells, promoted their apoptosis and induced G0/G1 cell cycle arrest. At the same time, PAUF-siRNA inhibited the invasion, adhesion and migration of the tumor cells. In conclusion, this study suggested that PAUF was expressed in CRC at a high frequency. Interference of PAUF may be an effective strategy for regulating malignant phenotypes of CRC through the Wnt/β-catenin pathway.

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1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
2	He SB, Yuan Y, Wang L, Yu MJ, Zhu YB and Zhu XG: Effects of cyclin-dependent kinase 8 specific siRNA on the proliferation and apoptosis of colon cancer cells. J Exp Clin Cancer Res. 30:1092011. View Article : Google Scholar : PubMed/NCBI
3	Wan D, He S, Xie B, Xu G, Gu W, Shen C, Hu Y, Wang X, Zhi Q and Wang L: Aberrant expression of miR-199a-3p and its clinical significance in colorectal cancers. Med Oncol. 30:3782013. View Article : Google Scholar : PubMed/NCBI
4	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
5	Markowitz SD and Bertagnolli MM: Molecular origins of cancer: molecular basis of colorectal cancer. N Engl J Med. 361:2449–2460. 2009. View Article : Google Scholar : PubMed/NCBI
6	Walther A, Johnstone E, Swanton C, Midgley R, Tomlinson I and Kerr D: Genetic prognostic and predictive markers in colorectal cancer. Nat Rev Cancer. 9:489–499. 2009. View Article : Google Scholar : PubMed/NCBI
7	Chen HJ, Hsu LS, Shia YT, Lin MW and Lin CM: The β-catenin/TCF complex as a novel target of resveratrol in the Wnt/β-catenin signaling pathway. Biochem Pharmacol. 84:1143–1153. 2012.
8	Firestein R and Hahn WC: Revving the Throttle on an oncogene: CDK8 takes the driver seat. Cancer Res. 69:7899–7901. 2009. View Article : Google Scholar : PubMed/NCBI
9	Luu HH, Zhang R, Haydon RC, Rayburn E, Kang Q, Si W, Park JK, Wang H, Peng Y, Jiang W and He TC: Wnt/β-catenin signaling pathway as a novel cancer drug target. Curr Cancer Drug Targets. 4:653–671. 2004.
10	Dihlmann S and von Knebel Doeberitz M: Wnt/β-catenin-pathway as a molecular target for future anti-cancer therapeutics. Int J Cancer. 113:515–524. 2005.
11	Kundu JK, Choi KY and Surh YJ: β-catenin-mediated signaling: a novel molecular target for chemoprevention with anti-inflammatory substances. Biochim Biophys Acta. 1765:14–24. 2006.
12	Kim SA, Lee Y, Jung DE, Park KH, Park JY, Gang J, Jeon SB, Park EC, Kim YG, Lee B, Liu Q, Zeng W, Yeramilli S, Lee S, Koh SS and Song SY: Pancreatic adenocarcinoma up-regulated factor (PAUF), a novel up-regulated secretory protein in pancreatic ductal adenocarcinoma. Cancer Sci. 100:828–836. 2009. View Article : Google Scholar : PubMed/NCBI
13	Cho IR, Koh SS, Min HJ, Kim SJ, Lee Y, Park EH, Ratakorn S, Jhun BH, Oh S, Johnston RN and Chung YH: Pancreatic adenocarcinoma up-regulated factor (PAUF) enhances the expression of β-catenin, leading to a rapid proliferation of pancreatic cells. Exp Mol Med. 43:82–90. 2011.PubMed/NCBI
14	Bienz M and Clevers H: Linking colorectal cancer to Wnt signaling. Cell. 103:311–320. 2000. View Article : Google Scholar : PubMed/NCBI
15	Nelson WJ and Nusse R: Convergence of Wnt, β-catenin, and cadherin pathways. Science. 303:1483–1487. 2004.
16	Coluccia AM, Benati D, Dekhil H, De Filippo A, Lan C and Gambacorti-Passerini C: SKI-606 decreases growth and motility of colorectal cancer cells by preventing pp60(c-Src)-dependent tyrosine phosphorylation of β-catenin and its nuclear signaling. Cancer Res. 66:2279–2286. 2006.PubMed/NCBI
17	Seo JO, Han SI and Lim SC: Role of CDK8 and β-catenin in colorectal adenocarcinoma. Oncol Rep. 24:285–291. 2010.
18	Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, Guney I, Freed E, Ligon AH, Vena N, Ogino S, Chheda MG, Tamayo P, Finn S, Shrestha Y, Boehm JS, Jain S, Bojarski E, Mermel C, Barretina J, Chan JA, Baselga J, Tabernero J, Root DE, Fuchs CS, Loda M, Shivdasani RA, Meyerson M and Hahn WC: CDK8 is a colorectal cancer oncogene that regulates β-catenin activity. Nature. 455:547–551. 2008.PubMed/NCBI
19	Morris EJ, Ji JY, Yang F, Di Stefano L, Herr A, Moon NS, Kwon EJ, Haigis KM, Näär AM and Dyson NJ: E2F1 represses β-catenin transcription and is antagonized by both pRB and CDK8. Nature. 455:552–556. 2008.
20	Lee Y, Kim SJ, Park HD, Park EH, Huang SM, Jeon SB, Kim JM, Lim DS and Koh SS: PAUF functions in the metastasis of human pancreatic cancer cells and upregulates CXCR4 expression. Oncogene. 29:56–67. 2010. View Article : Google Scholar : PubMed/NCBI
21	Park HD, Lee Y, Oh YK, Jung JG, Park YW, Myung K, Kim KH, Koh SS and Lim DS: Pancreatic adenocarcinoma upregulated factor promotes metastasis by regulating TLR/CXCR4 activation. Oncogene. 30:201–211. 2011. View Article : Google Scholar : PubMed/NCBI
22	Lee YS, Kim SJ, Min HJ, Jo JY, Park EH and Koh SS: PAUF promotes adhesiveness of pancreatic cancer cells by modulating focal adhesion kinase. Exp Mol Med. 43:291–297. 2011. View Article : Google Scholar : PubMed/NCBI
23	Kim YH, Sung HJ, Kim S, Kim EO, Lee JW, Moon JY, Choi K, Jung JE, Lee Y, Koh SS, Rhee SG, Heo K and Kim IH: An RNA aptamer that specifically binds pancreatic adenocarcinoma up-regulated factor inhibits migration and growth of pancreatic cancer cells. Cancer Lett. 313:76–83. 2011. View Article : Google Scholar : PubMed/NCBI
24	Merritt WM, Bar-Eli M and Sood AK: The dicey role of dicer: implications for RNAi therapy. Cancer Res. 70:2571–2574. 2010. View Article : Google Scholar : PubMed/NCBI
25	Brummelkamp TR, Bemards R and Agami R: Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell. 2:243–247. 2002. View Article : Google Scholar : PubMed/NCBI