Lentivirus-mediated shRNA targeting of cyclin D1 enhances the chemosensitivity of human gastric cancer to 5-fluorouracil

Seo,Jin-Hee; Jeong,Eui-Suk; Lee,Kyoung-Sun; Heo,Seung-Ho; Jeong,Dong-Gu; Choi,Yang-Kyu

doi:10.3892/ijo.2013.2119

Lentivirus-mediated shRNA targeting of cyclin D1 enhances the chemosensitivity of human gastric cancer to 5-fluorouracil

Authors:
- Jin-Hee Seo
- Eui-Suk Jeong
- Kyoung-Sun Lee
- Seung-Ho Heo
- Dong-Gu Jeong
- Yang-Kyu Choi
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Affiliations: Department of Laboratory Animal Medicine, College of Veterinary Medicine, Konkuk University, Seoul 143-701, Republic of Korea
Published online on: October 3, 2013 https://doi.org/10.3892/ijo.2013.2119
Pages: 2007-2014

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Abstract

Gastric cancer is one of the major public health problems. Despite new chemotherapeutic treatments, the prognosis of gastric cancer remains poor. 5-Fluorouracil (5-FU) is used as a standard chemotherapy drug in gastric cancer. However, 5-FU resistance develops frequently and is a main cause of chemotherapy failure in human gastric cancer. Overexpression of cyclin D1 is related to rapid cell growth, a poor prognosis and increased chemoresistance in several types of cancers. In this study, we investigated whether treatment of gastric cancer cells with shRNA targeting cyclin D1 (ShCCND1) or 5-FU, alone or in combination, influences the activation of phosphorylated AKT (pAKT) and pNFκB, which are markers that are increased in 5-FU chemoresistance. We also investigated the effect of combined treatment with ShCCND1 and 5-FU on cell growth and chemosensitivity to 5-FU in the gastric cancer cell line AGS. The data showed that ShCCND1-mediated cyclin D1 downregulation in AGS cells significantly inhibited cell proliferation, cell mobility and clonogenicity. In addition, combined treatment with ShCCND1 and 5-FU significantly decreased the survival rate of AGS cells, compared to single-treatment with either agent. These results demonstrated that ShCCND1 increases 5-FU chemosensitivity, a conclusion that is also supported by the concomitant reduction in expression of pAKT and pNFκB, increase of G1 arrest and induction of apoptosis. Taken together, these data provide further evidence that therapeutic strategies targeting cyclin D1 may have the dual advantage of suppressing the growth of cancer cells, while enhancing their chemosensitivity.

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1.	Leung WK, Wu MS, Kakugawa Y, Kim JJ, Yeoh KG, Goh KL, Wu KC, Wu DC, Sollano J, Kachintorn U, Gotoda T, Lin JT, You WC, Ng EK and Sung JJ; Asia Pacific Working Group on Gastric C: Screening for gastric cancer in Asia: current evidence and practice. Lancet Oncol. 9:279–287. 2008. View Article : Google Scholar : PubMed/NCBI
2.	Ohtsu A: Chemotherapy for metastatic gastric cancer: past, present and future. J Gastroenterol. 43:256–264. 2008. View Article : Google Scholar : PubMed/NCBI
3.	Sakuramoto S, Sasako M, Yamaguchi T, Kinoshita T, Fujii M, Nashimoto A, Furukawa H, Nakajima T, Ohashi Y, Imamura H, Higashino M, Yamamura Y, Kurita A and Arai K: Adjuvant chemotherapy for gastric cancer with S-1, an oral fluoropyrimidine. N Engl J Med. 357:1810–1820. 2007. View Article : Google Scholar : PubMed/NCBI
4.	GASTRIC (Global Advanced/Adjuvant Stomach Tumor Research International Collaboration) Group; Paoletti X, Oba K, Burzykowski T, Michiels S, Ohashi Y, Pignon JP, Rougier P, Sakamoto J, Sargent D, Sasako M, Van Cutsem E and Buyse M: Benefit of adjuvant chemotherapy for resectable gastric cancer: a meta-analysis. JAMA. 303:1729–1737. 2010. View Article : Google Scholar : PubMed/NCBI
5.	Sherr CJ: G1 phase progression: cycling on cue. Cell. 79:551–555. 1994. View Article : Google Scholar : PubMed/NCBI
6.	Weinberg RA: The retinoblastoma protein and cell cycle control. Cell. 81:323–330. 1995. View Article : Google Scholar : PubMed/NCBI
7.	Gao P, Zhou GY, Liu Y, Li JS, Zhen JH and Yuan YP: Alteration of cyclin D1 in gastric carcinoma and its clinicopathologic significance. World J Gastroenterol. 10:2936–2939. 2004.PubMed/NCBI
8.	Motokura T and Arnold A: Cyclin D and oncogenesis. Curr Opin Genet Dev. 3:5–10. 1993. View Article : Google Scholar
9.	Coco Martin JM, Balkenende A, Verschoor T, Lallemand F and Michalides R: Cyclin D1 overexpression enhances radiation-induced apoptosis and radiosensitivity in a breast tumor cell line. Cancer Res. 59:1134–1140. 1999.PubMed/NCBI
10.	Han EK, Begemann M, Sgambato A, Soh JW, Doki Y, Xing WQ, Liu W and Weinstein IB: Increased expression of cyclin D1 in a murine mammary epithelial cell line induces p27kip1, inhibits growth and enhances apoptosis. Cell Growth Differ. 7:699–710. 1996.PubMed/NCBI
11.	Kuroda Y, Sakai A, Tsuyama N, Katayama Y, Munemasa S, Asaoku H, Okikawa Y, Nakaju N, Mizuno M, Ogawa K, Nishisaka T, Matsui H, Tanaka H and Kimura A: Ectopic cyclin D1 overexpression increases chemosensitivity but not cell proliferation in multiple myeloma. Int J Oncol. 33:1201–1213. 2008.PubMed/NCBI
12.	Kornmann M, Arber N and Korc M: Inhibition of basal and mitogen-stimulated pancreatic cancer cell growth by cyclin D1 antisense is associated with loss of tumorigenicity and potentiation of cytotoxicity to cisplatinum. J Clin Invest. 101:344–352. 1998. View Article : Google Scholar : PubMed/NCBI
13.	Warenius HM, Seabra LA and Maw P: Sensitivity to cis-diamminedichloroplatinum in human cancer cells is related to expression of cyclin D1 but not C-RAF-1 protein. Int J Cancer. 67:224–231. 1996. View Article : Google Scholar : PubMed/NCBI
14.	Simizu S, Shibasaki F and Osada H: Bcl-2 inhibits calcineurin-mediated Fas ligand expression in antitumor drug-treated baby hamster kidney cells. Jpn J Cancer Res. 91:706–714. 2000. View Article : Google Scholar : PubMed/NCBI
15.	Zhou X, Zhang Z, Yang X, Chen W and Zhang P: Inhibition of cyclin D1 expression by cyclin D1 shRNAs in human oral squamous cell carcinoma cells is associated with increased cisplatin chemosensitivity. Int J Cancer. 124:483–489. 2009. View Article : Google Scholar : PubMed/NCBI
16.	Sun Y, Luo D and Liao DJ: Cyclin D1 protein plays different roles in modulating chemoresponses in MCF7 and MDA-MB231 cells. J Carcinog. 11:122012. View Article : Google Scholar : PubMed/NCBI
17.	Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E, Frisch S and Reed JC: Regulation of cell death protease caspase-9 by phosphorylation. Science. 282:1318–1321. 1998. View Article : Google Scholar : PubMed/NCBI
18.	Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J and Greenberg ME: Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell. 96:857–868. 1999. View Article : Google Scholar : PubMed/NCBI
19.	Curnock AP, Logan MK and Ward SG: Chemokine signalling: pivoting around multiple phosphoinositide 3-kinases. Immunology. 105:125–136. 2002. View Article : Google Scholar : PubMed/NCBI
20.	Baldwin AS: Control of oncogenesis and cancer therapy resistance by the transcription factor NF-kappaB. J Clin Invest. 107:241–246. 2001. View Article : Google Scholar : PubMed/NCBI
21.	Ozes ON, Mayo LD, Gustin JA, Pfeffer SR, Pfeffer LM and Donner DB: NF-kappaB activation by tumour necrosis factor requires the Akt serine-threonine kinase. Nature. 401:82–85. 1999. View Article : Google Scholar : PubMed/NCBI
22.	Wang C-Y, Mayo MW and Baldwin AS: TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-kappaB. Science. 274:784–787. 1996. View Article : Google Scholar : PubMed/NCBI
23.	Camp ER, Li J, Minnich DJ, Brank A, Moldawer LL, MacKay SL and Hochwald SN: Inducible nuclear factor-kappaB activation contributes to chemotherapy resistance in gastric cancer. J Am Coll Surg. 199:249–258. 2004. View Article : Google Scholar : PubMed/NCBI
24.	Lu YS, Yeh PY, Chuang SE, Gao M, Kuo ML and Cheng AL: Glucocorticoids enhance cytotoxicity of cisplatin via suppression of NF-κB activation in the glucocorticoid receptor-rich human cervical carcinoma cell line SiHa. J Endocrinol. 188:311–319. 2006.PubMed/NCBI
25.	Sauter ER, Nesbit M, Litwin S, Klein-Szanto AJ, Cheffetz S and Herlyn M: Antisense cyclin D1 induces apoptosis and tumor shrinkage in human squamous carcinomas. Cancer Res. 59:4876–4881. 1999.PubMed/NCBI
26.	Huang WS, Wang JP, Wang T, Fang JY, Lan P and Ma JP: ShRNA-mediated gene silencing of beta-catenin inhibits growth of human colon cancer cells. World J Gastroenterol. 13:6581–6587. 2007. View Article : Google Scholar : PubMed/NCBI
27.	Liang CC, Park AY and Guan JL: In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2:329–333. 2007. View Article : Google Scholar : PubMed/NCBI
28.	Tiang JM, Butcher NJ and Minchin RF: Small molecule inhibition of arylamine N-acetyltransferase Type I inhibits proliferation and invasiveness of MDA-MB-231 breast cancer cells. Biochem Biophys Res Commun. 393:95–100. 2010. View Article : Google Scholar : PubMed/NCBI
29.	Pearson G, English JM, White MA and Cobb MH: ERK5 and ERK2 cooperate to regulate NF-kappaB and cell transformation. J Biol Chem. 276:7927–7931. 2001. View Article : Google Scholar : PubMed/NCBI
30.	Martin V, Herrera F, Carrera-Gonzalez P, Garcia-Santos G, Antolin I, Rodriguez-Blanco J and Rodriguez C: Intracellular signaling pathways involved in the cell growth inhibition of glioma cells by melatonin. Cancer Res. 66:1081–1088. 2006. View Article : Google Scholar : PubMed/NCBI
31.	Cheng JC, Chou CH, Kuo ML and Hsieh CY: Radiation-enhanced hepatocellular carcinoma cell invasion with MMP-9 expression through PI3K/Akt/NF-kappaB signal transduction pathway. Oncogene. 25:7009–7018. 2006. View Article : Google Scholar : PubMed/NCBI
32.	Shin JY, Kim JO, Lee SK, Chae HS and Kang JH: LY294002 may overcome 5-FU resistance via down-regulation of activated p-AKT in Epstein-Barr virus-positive gastric cancer cells. BMC Cancer. 10:4252010. View Article : Google Scholar : PubMed/NCBI
33.	Kim TW, Kang YK, Ahn JH, Chang HM, Yook JH, Oh ST, Kim BS and Lee JS: Phase II study of capecitabine plus cisplatin as first-line chemotherapy in advanced gastric cancer. Ann Oncol. 13:1893–1898. 2002. View Article : Google Scholar : PubMed/NCBI
34.	Song M, Zhang R, Dai Y, Gao F, Chi H, Lv G, Chen B and Wang X: The in vitro inhibition of multidrug resistance by combined nanoparticulate titanium dioxide and UV irradition. Biomaterials. 27:4230–4238. 2006. View Article : Google Scholar : PubMed/NCBI
35.	Yoshida K and Toge T: Combination chemotherapy of TS-1 and docetaxel on advanced and recurrent gastric cancer. Gan To Kagaku Ryoho. 31:1982–1986. 2004.PubMed/NCBI
36.	Arlt A, Gehrz A, Muerkoster S, Vorndamm J, Kruse ML, Folsch UR and Schafer H: Role of NF-kappaB and Akt/PI3K in the resistance of pancreatic carcinoma cell lines against gemcitabine-induced cell death. Oncogene. 22:3243–3251. 2003. View Article : Google Scholar : PubMed/NCBI
37.	Fu M, Wang C, Li Z, Sakamaki T and Pestell RG: Minireview: Cyclin D1: normal and abnormal functions. Endocrinology. 145:5439–5447. 2004. View Article : Google Scholar : PubMed/NCBI
38.	Kornmann M, Danenberg KD, Arber N, Beger HG, Danenberg PV and Korc M: Inhibition of cyclin D1 expression in human pancreatic cancer cells is associated with increased chemosensitivity and decreased expression of multiple chemoresistance genes. Cancer Res. 59:3505–3511. 1999.
39.	Zhou H, Fujigaki Y, Kato A, Miyaji T, Yasuda H, Tsuji T, Yamamoto T, Yonemura K and Hishida A: Inhibition of p21 modifies the response of cortical proximal tubules to cisplatin in rats. Am J Physiol Renal Physiol. 291:F225–F235. 2006. View Article : Google Scholar : PubMed/NCBI
40.	Liu N, Zhang J, Zhang J, Liu S, Liu Y and Zheng D: Erbin-regulated sensitivity of MCF-7 breast cancer cells to TRAIL via ErbB2/AKT/NF-kappaB pathway. J Biochem. 143:793–801. 2008. View Article : Google Scholar : PubMed/NCBI