• 1.江蘇省連云港市第一人民醫(yī)院東方醫(yī)院普外科（連云港 222042）;;
  • 2.江蘇省南通市第三人民醫(yī)院普外科（南通 226000）;

目的  觀察小干擾RNA（small interference RNA, siRNA）對肝癌HepG2細(xì)胞血管內(nèi)皮生長因子（VEGF）表達(dá)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(xì)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(dá)，ELISA檢測細(xì)胞培養(yǎng)上清液中VEGF蛋白濃度。
結(jié)果  細(xì)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(xì)胞中VEGF mRNA和蛋白的表達(dá)明顯減少，細(xì)胞培養(yǎng)上清液中VEGF蛋白表達(dá)下降。
結(jié)論  運(yùn)用RNA干擾技術(shù)，可以有效地干擾肝癌HepG2細(xì)胞VEGF的表達(dá)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(xì)胞VEGF表達(dá)的研究. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(8): 633-636. doi: 復(fù)制

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(dá)載體對HepG2細(xì)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.
  1. 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. 2. Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors [J]. Nat Med, 2003; 9(6): 669-676.
  3. 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. 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. 5. 鄧偉， 梁力建. 脂質(zhì)體Tie-2-siRNA表達(dá)載體對HepG2細(xì)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. 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. 7. Folkman J. Clinical application of research of angiogenesis [J]. N Engl J Med, 1995; 333(26): 1757-1763.
  8. 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. 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. 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. 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.