Suppression of VEGF siRNA on Growth of Human Pancreatic Cancer Cells in Vitro_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

李善平 , 刘海威 , 程乐 , 张虎 , 曹登科 , 李政

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Keywords：

胰腺癌; 血管内皮生长因子; 小干扰RNA

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目的了解载体携带的小干扰RNA（small interfering RNA, siRNA）对人胰腺癌细胞株PANC-1血管内皮生长因子（vascular endothelial growth factor, VEGF）表达的抑制作用以及对肿瘤细胞生长的影响。方法设计并合成2对针对VEGF的siRNA 真核表达载体（PU-VEGF-siRNA1 组和 PU-VEGF-siRNA2 组，简称为重组质粒组），经脂质体Lipofectamine 2000 转染PANC-1 细胞，经G418 筛选后获得稳定转染细胞株，空载体转染组作为实验对照组，未转染组作为空白对照组。采用MTT法检测转染后PANC-1 细胞的增殖，流式细胞仪检测转染后PANC-1 细胞的凋亡率和细胞周期，逆转录聚合酶链反应（RT-PCR）检测转染后VEGF mRNA 表达情况。结果与实验对照组和空白对照组比较，两重组质粒组PANC-1 细胞的增殖减慢，细胞凋亡率增加，VEGF mRNA 表达明显下降，差异均有统计学意义（P lt;0.05）；细胞周期结果显示，两重组质粒组S 期细胞所占比例较两对照组明显减少（P lt;0.05）。结论siRNA 能有效抑制胰腺癌细胞VEGF 的表达，并能在体外抑制人胰腺癌细胞株PANC-1 细胞的生长。

Citation： 李善平,刘海威,程乐,张虎,曹登科,李政. Suppression of VEGF siRNA on Growth of Human Pancreatic Cancer Cells in Vitro. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2011, 18(6): 651-654. doi: Copy

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9.	Murata M, Takanami T, Shimizu S, et al. Inhibition of ocular angiogenesis by diced small interfering RNAs (siRNAs) specific to vascular endothelial growth factor (VEGF) [J]. Curr Eye Res， 2006， 31(2): 171180.
10.	Lu PY, Xie FY, Woodle MC. Modulation of angiogenesis with siRNA inhibitors for novel therapeutics [J]. Trends Mol Med， 2005， 11(3): 104113.
11.	Deans TL, Cantor CR, Collins JJ. A tunable genetic switch based on RNAi and repressor proteins for regulating gene expression in mammalian cells [J]. Cell， 2007， 130(2): 363372.
12.	Wurst M, Robles A, Po J, et al. An RNAi screen of the RRMdomain proteins of Trypanosoma brucei [J]. Mol Biochem Parasitol， 2009， 163(1): 6165.
13.	Parker GS, Maity TS, Bass BL. dsRNA binding properties of RDE4 and TRBP reflect their distinct roles in RNAi [J]. J Mol Biol， 2008， 384(4): 967979.
14.	Colmenares SU, Buker SM, Buhler M, et al. Coupling of doublestranded RNA synthesis and siRNA generation in fission yeast RNAi [J]. Mol Cell， 2007， 27(3): 449461.
15.	Takei Y, Kadomatsu K, Yuzawa Y， et al. A Small interfering RNA targeting vascular endothelial growth factor as cancer therapeutics [J]. Cancer Res, 2004, 64(10): 33653370.
16.	褚衍六, 赵幼安, 顾晓萌，等. RNA干扰抑制胃癌VEGF表达的实验研究 [J]. 中国肿瘤生物治疗杂志, 2005, 12(4): 297299.

1. 姜晓兵, 赵洪洋, 周凤，等. RNA干扰真核表达载体pEGFPH1介导的血管内皮生长因子shRNA治疗人胶质瘤的实验研究 [J]. 中华医学杂志, 2005, 85(8): 547550.
2. Hetian L, Ping A, Shumei S, et al. A novel peptide isolated from a phage display library inhibits tumor growth and metastasis by blocking the binding of vascular endothelial growth factor to its kinase domain receptor [J]. J Biol Chem, 2002, 277(45): 4313743142.
3. Kwon HS, Shin HC, Kim JS. Suppression of vascular endothelial growth factor expression at the transcriptional and posttranscriptional levels [J]. Nucleic Acids Res, 2005, 33(8): e74.
4. Holash J， Maisonpierre PC， Compton D， et al. Vessel cooption，regression and growth in tumors mediated by angiopoietins and VEGF [J]. Science， 1999， 284（5422）： 19941998.
5. Rafii S. Circulating endothelial precursors：mystery，reality and promise [J]. J Clin Invest， 2000， 105（1）： 1719.
6. Guo X， Chen Y， Xu Z， et al． Prognostic significance of VEGFC expression in correlation with COX2， lymphatic microvessel density，and clinicopathologic characteristics in human nonsmall cell lung cancer [J]． Acta Biochim Biophys Sin(Shanghai)， 2009， 41(3)： 217222.
7. 王勇，江丰收，孙玉蓓，等．肺癌患者血清VEGF的表达及其临床意义 [J]．临床肺科杂志， 2007， 12(8)： 804806.
8. Wannenes F, Ciafre SA, Niola F, et al. Vectorbased RNA interference against vascular endothelial growth factorA significantly limits vascularization and growth of prostate cancer in vivo [J]. Cancer Gene Ther， 2005， 12(12): 926934.
9. Murata M, Takanami T, Shimizu S, et al. Inhibition of ocular angiogenesis by diced small interfering RNAs (siRNAs) specific to vascular endothelial growth factor (VEGF) [J]. Curr Eye Res， 2006， 31(2): 171180.
10. Lu PY, Xie FY, Woodle MC. Modulation of angiogenesis with siRNA inhibitors for novel therapeutics [J]. Trends Mol Med， 2005， 11(3): 104113.
11. Deans TL, Cantor CR, Collins JJ. A tunable genetic switch based on RNAi and repressor proteins for regulating gene expression in mammalian cells [J]. Cell， 2007， 130(2): 363372.
12. Wurst M, Robles A, Po J, et al. An RNAi screen of the RRMdomain proteins of Trypanosoma brucei [J]. Mol Biochem Parasitol， 2009， 163(1): 6165.
13. Parker GS, Maity TS, Bass BL. dsRNA binding properties of RDE4 and TRBP reflect their distinct roles in RNAi [J]. J Mol Biol， 2008， 384(4): 967979.
14. Colmenares SU, Buker SM, Buhler M, et al. Coupling of doublestranded RNA synthesis and siRNA generation in fission yeast RNAi [J]. Mol Cell， 2007， 27(3): 449461.
15. Takei Y, Kadomatsu K, Yuzawa Y， et al. A Small interfering RNA targeting vascular endothelial growth factor as cancer therapeutics [J]. Cancer Res, 2004, 64(10): 33653370.
16. 褚衍六, 赵幼安, 顾晓萌，等. RNA干扰抑制胃癌VEGF表达的实验研究 [J]. 中国肿瘤生物治疗杂志, 2005, 12(4): 297299.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Suppression of VEGF siRNA on Growth of Human Pancreatic Cancer Cells in Vitro

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