Lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines significantly inhibits cell proliferation and tumor growth

Habib,Raees; Akhtar,Javed; Taqi,Mohammad; Yu,Che; Zhang,Chunqing

doi:10.3892/or.2015.4033

Lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines significantly inhibits cell proliferation and tumor growth

Authors:
- Raees Habib
- Javed Akhtar
- Mohammad Taqi
- Che Yu
- Chunqing Zhang
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Affiliations: Department of Gastroenterology, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China, Department of Thoracic Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China, Department of General Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 250012, P.R. China, Department of Nephrology, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China
Published online on: June 5, 2015 https://doi.org/10.3892/or.2015.4033
Pages: 859-867

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Abstract

It has been well documented that survivin has multiple functions including cytoprotection, inhibition of cell death, and cell cycle regulation, particularly at the mitotic stage of the cell cycle, all of which favor cancer survival. Its expression in normal tissue is developmentally regulated, and any type of deregulation in survivin expression favors cancer survival. Gastric cancer is one of the most common malignancies and the second most common cause of cancer-related mortality worldwide. The molecular mechanisms involved in the transformation and progression of gastric cancer remain unclear. In the present study, we investigated the effect of lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines. Lentiviral-mediated survivin shRNA was used to knock down survivin expression in gastric cancer cell lines SGC-7901, MGC-803 and MKN-28. The Τranswell chemotaxis and the CCK-8 assays were used to assess the migration and proliferation of the tumor cells, respectively. TUNEL assay was used to detect apoptosis. Quantitative real-time PCR and western blot analysis were used to quantify mRNA and protein levels, respectively. Our results demonstrated that lentiviral-mediated RNAi markedly suppressed the survivin expression in all three gastric cancer cell lines. Significant decrease in survivin mRNA and protein expression were detected in the gastric cancer cell lines stably transfected with the lentiviral survivin shRNA vector, and knockdown of survivin also significantly inhibited the proliferation and migration in the gastric cancer cells and tumorigenicity in a xenograft animal model. Our results indicated that aberrant high cytoplasmic survivin expression in gastric cancer cells is associated with increased proliferation index and tumor growth. In conclusion, our results suggest that lentiviral-mediated gene therapy has the potential to be developed into a novel therapeutic strategy for the treatment of gastric cancer.

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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 : PubMed/NCBI
2	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2013. CA Cancer J Clin. 63:11–30. 2013. View Article : Google Scholar : PubMed/NCBI
3	Yamamoto H, Ngan CY and Monden M: Cancer cells survive with survivin. Cancer Sci. 99:1709–1714. 2008. View Article : Google Scholar : PubMed/NCBI
4	Ambrosini G, Adida C and Altieri DC: A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med. 3:917–921. 1997. View Article : Google Scholar : PubMed/NCBI
5	Li F, Ambrosini G, Chu EY, Plescia J, Tognin S, Marchisio PC and Altieri DC: Control of apoptosis and mitotic spindle checkpoint by survivin. 396:580–584. 1998.
6	Jäättelä M: Escaping cell death: Survival proteins in cancer. Exp Cell Res. 248:30–43. 1999. View Article : Google Scholar : PubMed/NCBI
7	Adida C, Berrebi D, Peuchmaur M, Reyes-Mugica M and Altieri DC: Anti-apoptosis gene, survivin, and prognosis of neuroblastoma. Lancet. 351:882–883. 1998. View Article : Google Scholar : PubMed/NCBI
8	Kawasaki H, Altieri DC, Lu C-D, Toyoda M, Tenjo T and Tanigawa N: Inhibition of apoptosis by survivin predicts shorter survival rates in colorectal cancer. Cancer Res. 58:5071–5074. 1998.PubMed/NCBI
9	Monzó M, Rosell R, Felip E, Astudillo J, Sánchez JJ, Maestre J, Martín C, Font A, Barnadas A and Abad A: A novel anti-apoptosis gene: Re-expression of survivin messenger RNA as a prognosis marker in non-small-cell lung cancers. J Clin Oncol. 17:2100–2104. 1999.PubMed/NCBI
10	Swana HS, Grossman D, Anthony JN, Weiss RM and Altieri DC: Tumor content of the antiapoptosis molecule survivin and recurrence of bladder cancer. N Engl J Med. 341:452–453. 1999. View Article : Google Scholar : PubMed/NCBI
11	Sarela AI, Macadam RC, Farmery SM, Markham AF and Guillou PJ: Expression of the antiapoptosis gene, survivin, predicts death from recurrent colorectal carcinoma. Gut. 46:645–650. 2000. View Article : Google Scholar : PubMed/NCBI
12	Kato J, Kuwabara Y, Mitani M, Shinoda N, Sato A, Toyama T, Mitsui A, Nishiwaki T, Moriyama S, Kudo J, et al: Expression of survivin in esophageal cancer: Correlation with the prognosis and response to chemotherapy. Int J Cancer. 95:92–95. 2001. View Article : Google Scholar : PubMed/NCBI
13	Sohn DM, Kim SY, Baek MJ, Lim CW, Lee MH, Cho MS and Kim TY: Expression of survivin and clinical correlation in patients with breast cancer. Biomed Pharmacother. 60:289–292. 2006. View Article : Google Scholar : PubMed/NCBI
14	Shin S, Sung B-J, Cho Y-S, Kim HJ, Ha NC, Hwang JI, Chung CW, Jung YK and Oh BH: An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7. Biochemistry. 40:1117–1123. 2001. View Article : Google Scholar : PubMed/NCBI
15	Adamkov M, Kajo K, Vybohova D, Krajcovic J, Stuller F and Rajcani J: Correlations of survivin expression with clinicomor-phological parameters and hormonal receptor status in breast ductal carcinoma. Neoplasma. 59:30–37. 2012. View Article : Google Scholar
16	Sui L, Dong Y, Ohno M, Watanabe Y, Sugimoto K and Tokuda M: Survivin expression and its correlation with cell proliferation and prognosis in epithelial ovarian tumors. Int J Oncol. 21:315–320. 2002.PubMed/NCBI
17	Tanaka K, Iwamoto S, Gon G, Nohara T, Iwamoto M and Tanigawa N: Expression of survivin and its relationship to loss of apoptosis in breast carcinomas. Clin Cancer Res. 6:127–134. 2000.PubMed/NCBI
18	Zaffaroni N, Pennati M, Colella G, Perego P, Supino R, Gatti L, Pilotti S, Zunino F and Daidone MG: Expression of the anti-apoptotic gene survivin correlates with taxol resistance in human ovarian cancer. Cell Mol Life Sci. 59:1406–1412. 2002. View Article : Google Scholar : PubMed/NCBI
19	Mehrotra S, Languino LR, Raskett CM, Mercurio AM, Dohi T and Altieri DC: IAP regulation of metastasis. Cancer Cell. 17:53–64. 2010. View Article : Google Scholar : PubMed/NCBI
20	Yano J, Hirabayashi K, Nakagawa S, Yamaguchi T, Nogawa M, Kashimori I, Naito H, Kitagawa H, Ishiyama K, Ohgi T, et al: Antitumor activity of small interfering RNA/cationic liposome complex in mouse models of cancer. Clin Cancer Res. 10:7721–7726. 2004. View Article : Google Scholar : PubMed/NCBI
21	Akhtar J, Wang Z, Yu C and Zhang ZP: Effectiveness of local injection of lentivirus-delivered stathmin1 and stathmin1 shRNA in human gastric cancer xenograft mouse. J Gastroenterol Hepatol. 29:1685–1691. 2014. View Article : Google Scholar : PubMed/NCBI
22	Akhtar J, Wang Z, Zhang ZP and Bi MM: Lentiviral-mediated RNA interference targeting stathmin1 gene in human gastric cancer cells inhibits proliferation in vitro and tumor growth in vivo. J Transl Med. 11:2122013. View Article : Google Scholar : PubMed/NCBI
23	Chawla-Sarkar M, Bae SI, Reu FJ, Jacobs BS, Lindner DJ and Borden EC: Downregulation of Bcl-2, FLIP or IAPs (XIAP and survivin) by siRNAs sensitizes resistant melanoma cells to Apo2L/TRAIL-induced apoptosis. Cell Death Differ. 11:915–923. 2004. View Article : Google Scholar : PubMed/NCBI
24	Altieri DC: Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer. 8:61–70. 2008. View Article : Google Scholar
25	Lens SMA, Wolthuis RMF, Klompmaker R, Kauw J, Agami R, Brummelkamp T, Kops G and Medema RH: Survivin is required for a sustained spindle checkpoint arrest in response to lack of tension. EMBO J. 22:2934–2947. 2003. View Article : Google Scholar : PubMed/NCBI
26	Yang D, Welm A and Bishop JM: Cell division and cell survival in the absence of survivin. Proc Natl Acad Sci USA. 101:15100–15105. 2004. View Article : Google Scholar : PubMed/NCBI
27	Altieri DC: Validating survivin as a cancer therapeutic target. Nat Rev Cancer. 3:46–54. 2003. View Article : Google Scholar : PubMed/NCBI
28	Gu CM, Zhu YK, Ma YH, Zhang M, Liao B, Wu HY and Lin HL: Knockdown of survivin gene by vector-based short hairpin RNA technique induces apoptosis and growth inhibition in Burkitt’s lymphoma Raji cell line. Neoplasma. 53:206–212. 2006.
29	Yang J, Ouyang J, Ouyang L, Ouyang L and Chen Y: Inhibition of cell proliferation and increase of chemosensitivity by simultaneous knockdown of XIAP and survivin in pancreatic carcinoma cells. Oncol Res. 21:43–50. 2013. View Article : Google Scholar : PubMed/NCBI
30	Tamm I, Wang Y, Sausville E, Scudiero DA, Vigna N, Oltersdorf T and Reed JC: IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs. Cancer Res. 58:5315–5320. 1998.PubMed/NCBI
31	Guan HT, Xue XH, Wang XJ, Li A and Qin ZY: Knockdown of survivin expression by small interfering RNA suppresses proliferation of two human cancer cell lines. Chin Med Sci J. 21:115–119. 2006.PubMed/NCBI