Establishment of Hepatocellular Carcinoma Cell Lines with Stable Decreased Expressing IGF1R Gene with siRNA_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

NIU Jian ¹ , LI Xiangnong ¹ , HAN Zeguang ²

Department of General Surgery, The First Affiliated Hospital, Xuzhou Medical College, Xuzhou 221002, China;

Corresponding author：

Keywords：

Human insulin like growth factor-1 receptor; Small interfering RNA; Hepatocellular carcinoma; Gene transfection; Stable expression

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Objective 　To establish hepatocellular carcinoma (HCC) cell lines which olig-expressed IGF1R gene stably．
Methods 　An eukaryotic expressing vector pSUPER-IGF1R-siRNA that could block IGF1R expressing was transferred into hepatocellular carcinoma cell lines SMMC7721 and Hep3B with Lipofectamine 2000 reagents． After transferred, cells were selected with G418 to obtain positive clones. The expressions of IGF1R, cyclin D1 and cyclin B1 were detected by RT-PCR and Western-blot． Cell growth curve were painted.
Results 　Two cell lines clones were screened olig-expressing IGF1R gene stably． The experimental cell lines grew more slowly than control cell lines and the expression of cyclin D1 decreased (P＜0.05).
Conclusion 　The HCC cell lines for olig-expressing IGF1R gene stably are established successfully．The plasmid pSUPER-IGF1R-siRNA can inhibit the growth of SMMC7721 and Hep3B cell lines, and the expression of cyclin D1.

Citation： NIU Jian,LI Xiangnong,HAN Zeguang. Establishment of Hepatocellular Carcinoma Cell Lines with Stable Decreased Expressing IGF1R Gene with siRNA. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2007, 14(6): 679-684. doi: Copy

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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.	吕伟，张超. RNAi 在大肠癌基因治疗中的应用策略和展望 [J]. 中国普外基础与临床杂志, 2005; 12（6）∶637.
16.	陈华，景在平，包俊敏，等. RNA干扰对兔血管平滑肌细胞bcl2基因表达的影响 [J]. 中国普外基础与临床杂志, 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. 　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. 　牛坚, 钱海鑫，李向农，等. RNA干涉胰岛素样生长因子1类受体的研究 [J]. 中华实验外科杂志， 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. 　吕伟，张超. RNAi 在大肠癌基因治疗中的应用策略和展望 [J]. 中国普外基础与临床杂志, 2005; 12（6）∶637.
16. 　陈华，景在平，包俊敏，等. RNA干扰对兔血管平滑肌细胞bcl2基因表达的影响 [J]. 中国普外基础与临床杂志, 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.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Establishment of Hepatocellular Carcinoma Cell Lines with Stable Decreased Expressing IGF1R Gene with siRNA

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