• 三峽大學(xué)仁和醫(yī)院普通外科（湖北宜昌 443001）;

目的了解載體攜帶的小干擾RNA（small interfering RNA, siRNA）對人胰腺癌細(xì)胞株P(guān)ANC-1血管內(nèi)皮生長因子（vascular endothelial growth factor, VEGF）表達(dá)的抑制作用以及對腫瘤細(xì)胞生長的影響。方法 設(shè)計(jì)并合成2對針對VEGF的siRNA 真核表達(dá)載體（PU-VEGF-siRNA1 組和 PU-VEGF-siRNA2 組，簡稱為重組質(zhì)粒組），經(jīng)脂質(zhì)體Lipofectamine 2000 轉(zhuǎn)染PANC-1 細(xì)胞，經(jīng)G418 篩選后獲得穩(wěn)定轉(zhuǎn)染細(xì)胞株，空載體轉(zhuǎn)染組作為實(shí)驗(yàn)對照組，未轉(zhuǎn)染組作為空白對照組。采用MTT法檢測轉(zhuǎn)染后PANC-1 細(xì)胞的增殖，流式細(xì)胞儀檢測轉(zhuǎn)染后PANC-1 細(xì)胞的凋亡率和細(xì)胞周期，逆轉(zhuǎn)錄聚合酶鏈反應(yīng)（RT-PCR）檢測轉(zhuǎn)染后VEGF mRNA 表達(dá)情況。結(jié)果 與實(shí)驗(yàn)對照組和空白對照組比較，兩重組質(zhì)粒組PANC-1 細(xì)胞的增殖減慢，細(xì)胞凋亡率增加，VEGF mRNA 表達(dá)明顯下降，差異均有統(tǒng)計(jì)學(xué)意義（P lt;0.05）； 細(xì)胞周期結(jié)果顯示，兩重組質(zhì)粒組S 期細(xì)胞所占比例較兩對照組明顯減少（P lt;0.05）。 結(jié)論siRNA 能有效抑制胰腺癌細(xì)胞VEGF 的表達(dá)，并能在體外抑制人胰腺癌細(xì)胞株P(guān)ANC-1 細(xì)胞的生長。

引用本文： 李善平,劉海威,程樂,張虎,曹登科,李政. VEGF siRNA 抑制人胰腺癌PANC-1 細(xì)胞體外生長的研究. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(6): 651-654. doi: 復(fù)制

1. 姜曉兵, 趙洪洋, 周鳳， 等. RNA干擾真核表達(dá)載體pEGFPH1介導(dǎo)的血管內(nèi)皮生長因子shRNA治療人膠質(zhì)瘤的實(shí)驗(yàn)研究 [J]. 中華醫(yī)學(xué)雜志, 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， 江豐收， 孫玉蓓， 等． 肺癌患者血清VEGF的表達(dá)及其臨床意義 [J]． 臨床肺科雜志， 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, 趙幼安, 顧曉萌， 等. RNA干擾抑制胃癌VEGF表達(dá)的實(shí)驗(yàn)研究 [J]. 中國腫瘤生物治療雜志, 2005, 12(4): 297299.
  1. 1. 姜曉兵, 趙洪洋, 周鳳， 等. RNA干擾真核表達(dá)載體pEGFPH1介導(dǎo)的血管內(nèi)皮生長因子shRNA治療人膠質(zhì)瘤的實(shí)驗(yàn)研究 [J]. 中華醫(yī)學(xué)雜志, 2005, 85(8): 547550.
  2. 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. 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. 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. 5. Rafii S. Circulating endothelial precursors：mystery，reality and promise [J]. J Clin Invest， 2000， 105（1）： 1719.
  6. 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. 7. 王勇， 江豐收， 孫玉蓓， 等． 肺癌患者血清VEGF的表達(dá)及其臨床意義 [J]． 臨床肺科雜志， 2007， 12(8)： 804806.
  8. 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. 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. 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. 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. 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. 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. 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. 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. 16. 褚衍六, 趙幼安, 顧曉萌， 等. RNA干擾抑制胃癌VEGF表達(dá)的實(shí)驗(yàn)研究 [J]. 中國腫瘤生物治療雜志, 2005, 12(4): 297299.