目的 觀察小干擾RNA(small interference RNA, siRNA)對肝癌HepG2細胞血管內(nèi)皮生長因子(VEGF)表達的抑制作用。
方法 將VEGF siRNA經(jīng)LipofectamineTM 2000脂質(zhì)體轉(zhuǎn)染肝癌HepG2細胞后,實時熒光定量PCR和Western blot方法分別檢測VEGF mRNA和蛋白的表達,ELISA檢測細胞培養(yǎng)上清液中VEGF蛋白濃度。
結(jié)果 細胞培養(yǎng)6 h后siRNA的轉(zhuǎn)染效率為(90.4±2.9)%,轉(zhuǎn)染后肝癌HepG2細胞中VEGF mRNA和蛋白的表達明顯減少,細胞培養(yǎng)上清液中VEGF蛋白表達下降。
結(jié)論 運用RNA干擾技術(shù),可以有效地干擾肝癌HepG2細胞VEGF的表達并能降低VEGF的生成。
引用本文: 朱洪波,陳兆永,王勇,周曉春,王建新. 小干擾RNA抑制肝癌HepG2細胞VEGF表達的研究. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(8): 633-636. doi: 復制
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2. | Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors [J]. Nat Med, 2003; 9(6): 669-676. |
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4. | Semizarov D, Frost L, Sarthy A, et al. Specificity of short interfering RNA determined through gene expression signatures [J]. Proc Natl Acad Sci USA, 2003; 100(11): 6347-6352. |
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6. | Cruz PE, Mueller C, Cossette TL, et al. In vivo post-transcriptional gene silencing of alpha-1 antitrypsin by adeno-associated virus vectors expressing siRNA [J]. Lab Invest, 2007; 87(9): 893-902. |
7. | Folkman J. Clinical application of research of angiogenesis [J]. N Engl J Med, 1995; 333(26): 1757-1763. |
8. | Carpenter B, Lin Y, Stoll S, et al. VEGF is crucial for the hepatic vascular development required for lipoprotein uptake [J]. Development, 2005; 132(14): 3293-3303. |
9. | Yamaguchi R, Yano H, Iemura A, et al. Expression of vascular endothelial growth factor in human hepatocellular carcinoma [J]. Hepatology, 1998; 28(1): 68-77. |
10. | Poon RT, Lau C, Yu WC, et al. High serum levels of vascular endothelial growth factor predict poor response to transarterial chemoembolization in hepatocellular carcinoma: a prospective study [J]. Oncol Rep, 2004; 11(5): 1077-1084. |
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- 1. Elbashir SM, Martinez J, Patkaniowska A, et al. Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate [J]. EMBO J, 2001; 20(23): 6877-6888.
- 2. Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors [J]. Nat Med, 2003; 9(6): 669-676.
- 3. Fire A, Xu S, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans [J].Nature, 1998; 391(6669): 806-811.
- 4. Semizarov D, Frost L, Sarthy A, et al. Specificity of short interfering RNA determined through gene expression signatures [J]. Proc Natl Acad Sci USA, 2003; 100(11): 6347-6352.
- 5. 鄧偉, 梁力建. 脂質(zhì)體Tie-2-siRNA表達載體對HepG2細胞裸鼠移植瘤作用的研究 [J]. 中國普外基礎(chǔ)與臨床雜志, 2007; 14(5): 543-546.
- 6. Cruz PE, Mueller C, Cossette TL, et al. In vivo post-transcriptional gene silencing of alpha-1 antitrypsin by adeno-associated virus vectors expressing siRNA [J]. Lab Invest, 2007; 87(9): 893-902.
- 7. Folkman J. Clinical application of research of angiogenesis [J]. N Engl J Med, 1995; 333(26): 1757-1763.
- 8. Carpenter B, Lin Y, Stoll S, et al. VEGF is crucial for the hepatic vascular development required for lipoprotein uptake [J]. Development, 2005; 132(14): 3293-3303.
- 9. Yamaguchi R, Yano H, Iemura A, et al. Expression of vascular endothelial growth factor in human hepatocellular carcinoma [J]. Hepatology, 1998; 28(1): 68-77.
- 10. Poon RT, Lau C, Yu WC, et al. High serum levels of vascular endothelial growth factor predict poor response to transarterial chemoembolization in hepatocellular carcinoma: a prospective study [J]. Oncol Rep, 2004; 11(5): 1077-1084.
- 11. 解乃昌, 李國威, 王志亮, 等. 抗血管內(nèi)皮生長因子抗體對實驗性肝癌的生長抑制作用 [J]. 中國普外基礎(chǔ)與臨床雜志, 2002; 9(4): 242-245.