• 1.徐州醫(yī)學院附屬第一醫(yī)院普外科（徐州221002）;;
  • 2.國家人類基因組南方研究中心（上海201203）;

目的 　運用siRNA技術在肝癌細胞株中建立穩(wěn)定低表達人胰島素樣生長因子1類受體（IGF1R）基因的細胞株。
方法 　構建包含封閉IGF1R基因的真核表達載體pSUPER-IGF1R-siRNA，轉染SMMC7721和Hep3B細胞，G418篩選表達穩(wěn)定的細胞株。通過RT-PCR、Western-blot分析與鑒定IGF1R mRNA和蛋白及cyclin D1、cyclin B1蛋白的表達，并繪制細胞生長曲線。
結果 　在SMMC7721和Hep3B細胞中成功建立低表達IGF1R基因的細胞株，其生長明顯減慢(P＜0.05),且其cyclin D1表達亦明顯下降(P＜0.05)。
結論 　pSUPER-IGF1R-siRNA能抑制SMMC7721和Hep3B細胞的生長。

引用本文： 牛堅,李向農,韓澤廣. 運用siRNA建立穩(wěn)定低表達IGF1R基因的肝癌細胞株的實驗研究. 中國普外基礎與臨床雜志, 2007, 14(6): 679-684. doi: 復制

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1. 　Russell WE, Van Wyk JJ, Pledger WJ. Inhibition of the mitogenic effects of plasma by a monoclonal antibody to somatomedin C [J]. Proc Natl Acad Sci USA, 1984; 81(8)∶2389.
2. 　Blakesley VA, Koval AP, Stannard BS, et al. Replacement of tyrosine 1251 in the carboxyl terminus of the insulin-like growth factor-Ⅰ receptor disrupts the actin cytoskeleton and inhibits proliferation and anchorage-independent growth [J]. J Biol Chem, 1998; 273(29)∶18411.
3. 　Valentinis B, Baserga R. IGF-Ⅰ receptor signalling in transformation and differentiation [J]. Mol Pathol, 2001; 54(3)∶133.
4. 　Hellawell GO, Turner GD, Davies DR, et al. Expression of the type 1 insulin-like growth factor receptor is up-regulated in primary prostate cancer and commonly persists in metastatic disease [J]. Cancer Res, 2002; 62(10)∶2942.
5. 　Brummelkamp TR, Bernards R, Agami R. A system for stable expression of short interfering RNAs in mammalian cells [J]. Science, 2002; 296(5567)∶550.
6. 　牛堅, 錢海鑫， 李向農， 等. RNA干涉胰島素樣生長因子1類受體的研究 [J]. 中華實驗外科雜志， 2006; 23(7)∶872.
7. 　Sachdev D, Li SL, Hartell JS, et al. A chimeric humanized single-chain antibody against the type Ⅰ insulin-like growth factor (IGF) receptor renders breast cancer cells refractory to the mitogenic effects of IGF-Ⅰ [J]. Cancer Res, 2003; 63(3)∶627.
8. 　Andrews DW, Resnicoff M, Flanders AE, et al. Results of a pilot study involving the use of an antisense oligodeoxynucleotide directed against the insulin-like growth factor type Ⅰ receptor in malignant astrocytomas [J]. J Clin Oncol, 2001; 19(8)∶2189.
9. 　Moschos SJ, Mantzoros CS. The role of the IGF system in cancer: from basic to clinical studies and clinical applications [J]. Oncology, 2002; 63(4)∶317.
10. 　Sell C, Dumenil G, Deveaud C, et al. Effect of a null mutation of the insulin-like growth factor Ⅰ receptor gene on growth and transformation of mouse embryo fibroblasts [J]. Mol Cell Biol, 1994; 14(6)∶3604.
11. 　Werner H. Dysregulation of the type 1 IGF receptor as a paradigm in tumor progression [J]. Mol Cell Endocrinol, 1998; 141(1-2)∶1.
12. 　Borkhardt A. Blocking oncogenes in malignant cells by RNA interference——new hope for a highly specific cancer treatment? [J]. Cancer Cell, 2002; 2(3)∶167.
13. 　Camirand A, Lu Y, Pollak M. Co-targeting HER2/ErbB2 and insulin-like growth factor-1 receptors causes synergistic inhibition of growth in HER2-overexpressing breast cancer cells [J]. Med Sci Monit, 2002; 8(12)∶BR521.
14. 　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.
15. 　呂偉， 張超. RNAi 在大腸癌基因治療中的應用策略和展望 [J]. 中國普外基礎與臨床雜志, 2005; 12（6）∶637.
16. 　陳華， 景在平， 包俊敏， 等. RNA干擾對兔血管平滑肌細胞bcl2基因表達的影響 [J]. 中國普外基礎與臨床雜志, 2007; 14（1）∶56.
17. 　Baserga R. The insulin-like growth factor Ⅰ receptor: a key to tumor growth? [J]. Cancer Res, 1995; 55(2)∶249.
18. 　Valentinis B, Baserga R. IGF-Ⅰ receptor signalling in transformation and differentiation [J]. Mol Pathol, 2001; 54(3)∶133.
19. 　Liu X, Turbyville T, Fritz A, et al. Inhibition of insulin-like growth factor Ⅰ receptor expression in neuroblastoma cells induces the regression of established tumors in mice [J]. Cancer Res， 1998; 58(23)∶5432.
  1. 1. 　Russell WE, Van Wyk JJ, Pledger WJ. Inhibition of the mitogenic effects of plasma by a monoclonal antibody to somatomedin C [J]. Proc Natl Acad Sci USA, 1984; 81(8)∶2389.
  2. 2. 　Blakesley VA, Koval AP, Stannard BS, et al. Replacement of tyrosine 1251 in the carboxyl terminus of the insulin-like growth factor-Ⅰ receptor disrupts the actin cytoskeleton and inhibits proliferation and anchorage-independent growth [J]. J Biol Chem, 1998; 273(29)∶18411.
  3. 3. 　Valentinis B, Baserga R. IGF-Ⅰ receptor signalling in transformation and differentiation [J]. Mol Pathol, 2001; 54(3)∶133.
  4. 4. 　Hellawell GO, Turner GD, Davies DR, et al. Expression of the type 1 insulin-like growth factor receptor is up-regulated in primary prostate cancer and commonly persists in metastatic disease [J]. Cancer Res, 2002; 62(10)∶2942.
  5. 5. 　Brummelkamp TR, Bernards R, Agami R. A system for stable expression of short interfering RNAs in mammalian cells [J]. Science, 2002; 296(5567)∶550.
  6. 6. 　牛堅, 錢海鑫， 李向農， 等. RNA干涉胰島素樣生長因子1類受體的研究 [J]. 中華實驗外科雜志， 2006; 23(7)∶872.
  7. 7. 　Sachdev D, Li SL, Hartell JS, et al. A chimeric humanized single-chain antibody against the type Ⅰ insulin-like growth factor (IGF) receptor renders breast cancer cells refractory to the mitogenic effects of IGF-Ⅰ [J]. Cancer Res, 2003; 63(3)∶627.
  8. 8. 　Andrews DW, Resnicoff M, Flanders AE, et al. Results of a pilot study involving the use of an antisense oligodeoxynucleotide directed against the insulin-like growth factor type Ⅰ receptor in malignant astrocytomas [J]. J Clin Oncol, 2001; 19(8)∶2189.
  9. 9. 　Moschos SJ, Mantzoros CS. The role of the IGF system in cancer: from basic to clinical studies and clinical applications [J]. Oncology, 2002; 63(4)∶317.
  10. 10. 　Sell C, Dumenil G, Deveaud C, et al. Effect of a null mutation of the insulin-like growth factor Ⅰ receptor gene on growth and transformation of mouse embryo fibroblasts [J]. Mol Cell Biol, 1994; 14(6)∶3604.
  11. 11. 　Werner H. Dysregulation of the type 1 IGF receptor as a paradigm in tumor progression [J]. Mol Cell Endocrinol, 1998; 141(1-2)∶1.
  12. 12. 　Borkhardt A. Blocking oncogenes in malignant cells by RNA interference——new hope for a highly specific cancer treatment? [J]. Cancer Cell, 2002; 2(3)∶167.
  13. 13. 　Camirand A, Lu Y, Pollak M. Co-targeting HER2/ErbB2 and insulin-like growth factor-1 receptors causes synergistic inhibition of growth in HER2-overexpressing breast cancer cells [J]. Med Sci Monit, 2002; 8(12)∶BR521.
  14. 14. 　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.
  15. 15. 　呂偉， 張超. RNAi 在大腸癌基因治療中的應用策略和展望 [J]. 中國普外基礎與臨床雜志, 2005; 12（6）∶637.
  16. 16. 　陳華， 景在平， 包俊敏， 等. RNA干擾對兔血管平滑肌細胞bcl2基因表達的影響 [J]. 中國普外基礎與臨床雜志, 2007; 14（1）∶56.
  17. 17. 　Baserga R. The insulin-like growth factor Ⅰ receptor: a key to tumor growth? [J]. Cancer Res, 1995; 55(2)∶249.
  18. 18. 　Valentinis B, Baserga R. IGF-Ⅰ receptor signalling in transformation and differentiation [J]. Mol Pathol, 2001; 54(3)∶133.
  19. 19. 　Liu X, Turbyville T, Fritz A, et al. Inhibition of insulin-like growth factor Ⅰ receptor expression in neuroblastoma cells induces the regression of established tumors in mice [J]. Cancer Res， 1998; 58(23)∶5432.