RNA interference‑mediated inhibition of survivin and VEGF in pancreatic cancer cells in vitro

Song,Jianlin; Cao,Lipin; Li,Yixiong

doi:10.3892/mmr.2013.1361

RNA interference‑mediated inhibition of survivin and VEGF in pancreatic cancer cells in vitro

Authors:
- Jianlin Song
- Lipin Cao
- Yixiong Li
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Affiliations: Department of General Surgery, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R. China
Published online on: March 6, 2013 https://doi.org/10.3892/mmr.2013.1361
Pages: 1651-1655

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Abstract

The aim of the present study was to investigate the effects of simultaneous short hairpin RNA (shRNA)‑targeted survivin and vascular endothelial growth factor (VEGF) inhibition on the proliferation, apoptosis and angiogenesis of human pancreatic cancer cells (Panc‑1). Targeted small interfering RNA (siRNA) expression vectors of survivin and VEGF were constructed and transfected into Panc‑1 cells. The downregulation of survivin and VEGF expression was evaluated by real‑time PCR and western blot analysis. The effects of targeted shRNA on the proliferation and apoptosis of Panc‑1 cells were analyzed by MTT assay and flow cytometry (FCM). The culture medium from Panc‑1 cells transfected with siRNA was collected and human umbilical vein endothelial cells (HUVECs) were seeded in this media. The proliferation and apoptosis of the HUVECs were also investigated by MTT assay and FCM. A transfected cell line (Panc‑1/survivin‑shRNA and Panc‑1/VEGF‑shRNA) was established in which the expression of survivin and VEGF was downregulated. The cell viabilities of Panc‑1 cells and HUVECs in the combined inhibition groups were markedly decreased compared with the controls. The cell apoptosis rates of Panc‑1 cells and HUVECs in the combined inhibition groups were observed to be significantly increased compared with the controls. The simultaneous RNA interference‑mediated downregulation of survivin and VEGF expression inhibited proliferation and induced the apoptosis of Panc‑1 cells and HUVECs, indicating that combined therapy with survivin and VEGF inhibition may serve as a potential strategy for the treatment of pancreatic cancer.

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1	Saif MW, Sviglin H and Carpenter M: Impact of ethnicity on outcome in pancreatic carcinoma. JOP. 6:246–254. 2005.PubMed/NCBI
2	Rodriguez JA, Li M, Yao Q, Chen C and Fisher WE: Gene overexpression in pancreatic adenocarcinoma: diagnostic and therapeutic implications. World J Surg. 29:297–305. 2005. View Article : Google Scholar : PubMed/NCBI
3	Kawasaki H, Toyoda M, Shinohara H, et al: Expression of survivin correlates with apoptosis, proliferation and angiogenesis during human colorectal tumorigenesis. Cancer. 91:2026–2032. 2001. View Article : Google Scholar : PubMed/NCBI
4	Ikeda N, Nakajima Y, Sho M, et al: The association of K-ras gene mutation and vascular endothelial growth factor gene expression in pancreatic carcinoma. Cancer. 92:488–499. 2001. View Article : Google Scholar : PubMed/NCBI
5	Satoh K, Kaneko K, Hirota M, Masamune A, Satoh A and Shimosegawa T: Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors. Cancer. 92:271–278. 2001. View Article : Google Scholar : PubMed/NCBI
6	Lee MA, Park GS, Lee HJ, et al: Survivin expression and its clinical significance in pancreatic cancer. BMC Cancer. 5:1272005. View Article : Google Scholar : PubMed/NCBI
7	Itakura J, Ishiwata T, Friess H, et al: Enhanced expression of vascular endothelial growth factor in human pancreatic cancer correlates with local disease progression. Clin Cancer Res. 3:1309–1316. 1997.
8	Ikeda N, Adachi M, Taki T, et al: Prognostic significance of angiogenesis in human pancreatic cancer. Br J Cancer. 79:1553–1563. 1999. View Article : Google Scholar : PubMed/NCBI
9	Olie RA, Simoes-Wust AP, Baumann B, et al: A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy. Cancer Res. 60:2805–2809. 2000.
10	Ciardiello F, Bianco R, Damiano V, et al: Antiangiogenic and antitumor activity of anti-epidermal growth factor receptor C225 monoclonal antibody in combination with vascular endothelial growth factor antisense oligonucleotide in human GEO colon cancer cells. Clin Cancer Res. 6:3739–3747. 2000.
11	Yonesaka K, Tamura K, Kurata T, et al: Small interfering RNA targeting survivin sensitizes lung cancer cell with mutant p53 to adriamycin. Int J Cancer. 118:812–820. 2006. View Article : Google Scholar : PubMed/NCBI
12	Mita AC, Mita MM, Nawrocki ST and Giles FJ: Survivin: key regulator of mitosis and apoptosis and novel target for cancer therapeutics. Clin Cancer Res. 14:5000–5005. 2008. View Article : Google Scholar : PubMed/NCBI
13	Dvorak HF, Nagy JA, Feng D, Brown LF and Dvorak AM: Vascular permeability factor/vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Microbiol Immunol. 237:97–132. 1999.
14	Jones A and Fujiyama C: Angiogenesis in urological malignancy: prognostic indicator and therapeutic target. BJU Int. 83:535–555. 1999. View Article : Google Scholar : PubMed/NCBI
15	Hayashibara T, Yamada Y, Miyanishi T, et al: Vascular endothelial growth factor and cellular chemotaxis: a possible autocrine pathway in adult T-cell leukemia cell invasion. Clin Cancer Res. 7:2719–2726. 2001.PubMed/NCBI
16	Xie K, Wei D and Huang S: Transcriptional anti-angiogenesis therapy of human pancreatic cancer. Cytokine Growth Factor Rev. 17:147–156. 2006. View Article : Google Scholar : PubMed/NCBI
17	Jiang C, Tan T, Yi XP, Shen H and Li YX: Lentivirus-mediated shRNA targeting XIAP and survivin inhibit SW1990 pancreatic cancer cell proliferation in vitro and in vivo. Mol Med Rep. 4:667–674. 2011.PubMed/NCBI
18	O’Connor DS, Schechner JS, Adida C, et al: Control of apoptosis during angiogenesis by survivin expression in endothelial cells. Am J Pathol. 156:393–398. 2000.
19	Mesri M, Morales-Ruiz M, Ackermann EJ, et al: Suppression of vascular endothelial growth factor-mediated endothelial cell protection by survivin targeting. Am J Pathol. 158:1757–1765. 2001. View Article : Google Scholar : PubMed/NCBI
20	Takei Y, Kadomatsu K, Yuzawa Y, Matsuo S and Muramatsu T: A small interfering RNA targeting vascular endothelial growth factor as cancer therapeutics. Cancer Res. 64:3365–3370. 2004. View Article : Google Scholar : PubMed/NCBI
21	Forster Y, Meye A, Krause S and Schwenzer B: Antisense-mediated VEGF suppression in bladder and breast cancer cells. Cancer Lett. 212:95–103. 2004. View Article : Google Scholar : PubMed/NCBI
22	Meister G, Landthaler M, Dorsett Y and Tuschl T: Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencing. RNA. 10:544–550. 2004. View Article : Google Scholar : PubMed/NCBI