Effect of Small Interfering RNA Targeting Inhibition of Growth Hormone Receptor on Proliferation and Invasion of Human Liver Cancer Line SMMC-7721_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 QIANChong-wei , QIHao-long , FENGMeng-long ,  LIUZhi-su

Department of General Surgery, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China;

Corresponding author：

LIUZhi-su, Email: 37609306@qq.com

Keywords：

Liver neoplasm; Growth hormone receptor; Small interfering RNA; Proliferation; Invasion

DOI：

10.7507/1007-9424.20160047

Video：

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Objective To investigate effect of small interfering RNA (siRNA) targeting inhibition of growth hormone receptor (GHR) on proliferation and invasion of human liver cancer line SMMC-7721. Methods SMMC-7721 cells were transfected with siRNA targeting human GHR by GenMute^TM transfection regent.The cells were divided into three groups:blank control group (non-transfected siRNA),negative control group (transfected with non-specific siRNA),and specificity transfected group (transfected with expression specifically interfere by GHR siRNA).the relative expression of GHR mRNA was detected by using real-time PCR.the expression of GHR protein was detected by using Western blot.The cell proliferation was assessed by CCK-8 assay.And the ability of invasion was examined by Transwell assay. Results The expressions of GHR mRNA and GHR protein in the specificity transfected group were significantly lower than those in the blank control group (P<0.05) and the negative control group (P<0.05).Compared with the blank control group and the negative control group,the absorbance value and the number of migrating cells of SMMC-7721 cells were decreased obviously (P<0.05) in the specificity transfected group. Conclusion siRNA targeting human GHR could reduce capability of proliferation migration and invasion of SMMC-7721 cells.

Citation： QIANChong-wei, QIHao-long, FENGMeng-long, LIUZhi-su. Effect of Small Interfering RNA Targeting Inhibition of Growth Hormone Receptor on Proliferation and Invasion of Human Liver Cancer Line SMMC-7721. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2016, 23(2): 177-181. doi: 10.7507/1007-9424.20160047 Copy

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2.	Perry JK, Emerald BS, Mertani HC, et al. The oncogenic potential of growth hormone. Growth Horm IGF Res, 2006, 16(5-6):277-289.
3.	周东, 张毅, 梁道明, 等. siRNA靶向抑制GHR对人结肠癌SW480细胞增殖的影响. 中国普通外科杂志, 2011, 20(9):945-950.
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5.	吴孟超, 沈锋, 傅晓辉. 中国肝脏外科的发展之路. 中华实验外科杂志, 2010, 27(12):1765-1766.
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7.	Turner HE, Harris AL, Melmed S, et al. Angiogenesis in endocrine tumors. Endocr Rev, 2003, 24(5):600-632.
8.	Blum H. Hepatocellular carcinoma:therapy and prevention. World J Gastroenterol, 2005, 11(47):7391-7400.
9.	Zhu T, Starling-Emerald B, Zhang X, et al. Oncogenic transforma-tion of human mammaryepithelial cells by autocrine human growth hormone. Cancer Res, 2005, 65(1):317-324.
10.	Chang TC, Lin JJ, Yu SC, et al. Absence of growth-hormone receptor in hepatocellular carcinoma and cirrhotic liver. Hepatology, 1990, 11(1):123-126.
11.	Banerjee I, Clayton PE. Growth hormone treatment and cancer risk. Endocrinol Metab Clin North Am, 2007, 36(1):247-263.
12.	向春华, 严律南, 林琦远,等. 生长激素受体在人肝癌组织中的表达.中国普外基础与临床杂志, 2002, 9(4):255-257.
13.	Amit T, Hacham H, Daily O, et al. The HepG2 cell line in the study of growth hormone receptor/binding protein. Mol Cell EndocrinoI, 1994, 101(1-2):29-36.
14.	Pang RW, Joh JW, Johnson PJ, et al. Biology of hepatocellular carcinoma. Ann Surg Oncol, 2008, 15(4):962-971.
15.	Lan HN, Li W, Jiang HL, et al. Intracellular signaling transduction pathways triggered by a well-known anti-GHR monoclonal antibody, Mab263, in vitro and in vivo. Int J Mol Sci, 2014, 15(11):20538-20554.
16.	Hou G, Xue L, Lu Z, et al. An activated mTOR/p70S6K signaling pathway in esophageal squamous cell carcinoma cell lines and inhibition of the pathway by rapamycin and siRNA against mTOR. Cancer Lett, 2007, 253(2):236-248.
17.	Beentjes JA, van Gorkom BA, Sluiter WJ, et al. One year growth hormone replacement therapy does not alter colonic epithelial cell proliferation in growth hormone deficient adults. Clin Endocrinol (Oxf), 2000, 52(4):457-462.
18.	Yin DH, Vreeland F, Schaaf LJ, et al. Clinical pharmacodynamic effects of the growth hormone receptor antagonist pegvisomant:Implications for cancer therapy. Clin Cancer Res, 2007, 13(3):1000-1009.
19.	Lorenzini A, Salmon AB, Lerner C, et al. Mice producing reduced levels of insulin-like growth factor type 1 display an increase in maximum, but not mean, life span. J Gerontol A Biol Sci Med Sci, 2014, 69(4):410-419.
20.	Kesireddy V, Van Der Ven PF, Fuerst DO. Multipurpose modular lentiviral vectors for RNA interference and transgene expression. Mol Biol Rep, 2010, 37(6):2863-2870.
21.	Cao J, Luo SM, Liang L, et al. Effects of parenteral nutrition without and with growth hormone on growth hormone/insulin-like growth factor-1 axis after hepatectomy in hepatocellular carcinoma with liver cirrhosis. JPEN J Parenter Enteral Nutr, 2007, 31(6):496-501.
22.	Stangelberger A, Schally AV, Varga JL, et al. Antagonists of growth hormone releasing hormone (GHRH) and of bombesin/gastrin releasing peptide (BN/GRP) suppress the expression of VEGF, bFGF, and receptors of the EGF/HER family in PC-3 and DU-145 human androgen-independent prostate cancers. Prostate, 2005, 64(3):303-315.
23.	Luo SM, Liang LJ, Lai JM. Effects of recombinant human growth hormone on remnant liver after hepatectomy in hepatocellular carcinoma with cirrhosis. World J Gastroenterol, 2004, 10(9):1292-1296.
24.	Shi FF, Li SY. Experimental study on effect of recombinant human growth hormone combined with chemotherapy on stomach neoplasms implanted in nude mice. Chinese-German J Clin Oncol, 2007, 6(1):27-31.
25.	Bartke A, Westbrook R, Sun L, et al. Links between growth hormone and aging. Endokrynol Pol, 2013, 64(1):46-52.

1. Scharf JG, Dombrowski F, Ramadori G. The IGF axis and hepato-carcinogenesis. Mol Pathol, 2001, 54(3):138-144.
2. Perry JK, Emerald BS, Mertani HC, et al. The oncogenic potential of growth hormone. Growth Horm IGF Res, 2006, 16(5-6):277-289.
3. 周东, 张毅, 梁道明, 等. siRNA靶向抑制GHR对人结肠癌SW480细胞增殖的影响. 中国普通外科杂志, 2011, 20(9):945-950.
4. Brooks AJ, Wooh JW, Tunny KA, et al. Growth hormone receptor; mechanism of action. Int Biochem Cell Biology. Int J Biochem Cell Biol, 2008, 40(10):1984-1989.
5. 吴孟超, 沈锋, 傅晓辉. 中国肝脏外科的发展之路. 中华实验外科杂志, 2010, 27(12):1765-1766.
6. Pang R, Poon RT. Angiogenesis and antiangiogenic therapy in hepatocellular carcinoma. Cancer Lett, 2006, 242(2):151-167.
7. Turner HE, Harris AL, Melmed S, et al. Angiogenesis in endocrine tumors. Endocr Rev, 2003, 24(5):600-632.
8. Blum H. Hepatocellular carcinoma:therapy and prevention. World J Gastroenterol, 2005, 11(47):7391-7400.
9. Zhu T, Starling-Emerald B, Zhang X, et al. Oncogenic transforma-tion of human mammaryepithelial cells by autocrine human growth hormone. Cancer Res, 2005, 65(1):317-324.
10. Chang TC, Lin JJ, Yu SC, et al. Absence of growth-hormone receptor in hepatocellular carcinoma and cirrhotic liver. Hepatology, 1990, 11(1):123-126.
11. Banerjee I, Clayton PE. Growth hormone treatment and cancer risk. Endocrinol Metab Clin North Am, 2007, 36(1):247-263.
12. 向春华, 严律南, 林琦远,等. 生长激素受体在人肝癌组织中的表达.中国普外基础与临床杂志, 2002, 9(4):255-257.
13. Amit T, Hacham H, Daily O, et al. The HepG2 cell line in the study of growth hormone receptor/binding protein. Mol Cell EndocrinoI, 1994, 101(1-2):29-36.
14. Pang RW, Joh JW, Johnson PJ, et al. Biology of hepatocellular carcinoma. Ann Surg Oncol, 2008, 15(4):962-971.
15. Lan HN, Li W, Jiang HL, et al. Intracellular signaling transduction pathways triggered by a well-known anti-GHR monoclonal antibody, Mab263, in vitro and in vivo. Int J Mol Sci, 2014, 15(11):20538-20554.
16. Hou G, Xue L, Lu Z, et al. An activated mTOR/p70S6K signaling pathway in esophageal squamous cell carcinoma cell lines and inhibition of the pathway by rapamycin and siRNA against mTOR. Cancer Lett, 2007, 253(2):236-248.
17. Beentjes JA, van Gorkom BA, Sluiter WJ, et al. One year growth hormone replacement therapy does not alter colonic epithelial cell proliferation in growth hormone deficient adults. Clin Endocrinol (Oxf), 2000, 52(4):457-462.
18. Yin DH, Vreeland F, Schaaf LJ, et al. Clinical pharmacodynamic effects of the growth hormone receptor antagonist pegvisomant:Implications for cancer therapy. Clin Cancer Res, 2007, 13(3):1000-1009.
19. Lorenzini A, Salmon AB, Lerner C, et al. Mice producing reduced levels of insulin-like growth factor type 1 display an increase in maximum, but not mean, life span. J Gerontol A Biol Sci Med Sci, 2014, 69(4):410-419.
20. Kesireddy V, Van Der Ven PF, Fuerst DO. Multipurpose modular lentiviral vectors for RNA interference and transgene expression. Mol Biol Rep, 2010, 37(6):2863-2870.
21. Cao J, Luo SM, Liang L, et al. Effects of parenteral nutrition without and with growth hormone on growth hormone/insulin-like growth factor-1 axis after hepatectomy in hepatocellular carcinoma with liver cirrhosis. JPEN J Parenter Enteral Nutr, 2007, 31(6):496-501.
22. Stangelberger A, Schally AV, Varga JL, et al. Antagonists of growth hormone releasing hormone (GHRH) and of bombesin/gastrin releasing peptide (BN/GRP) suppress the expression of VEGF, bFGF, and receptors of the EGF/HER family in PC-3 and DU-145 human androgen-independent prostate cancers. Prostate, 2005, 64(3):303-315.
23. Luo SM, Liang LJ, Lai JM. Effects of recombinant human growth hormone on remnant liver after hepatectomy in hepatocellular carcinoma with cirrhosis. World J Gastroenterol, 2004, 10(9):1292-1296.
24. Shi FF, Li SY. Experimental study on effect of recombinant human growth hormone combined with chemotherapy on stomach neoplasms implanted in nude mice. Chinese-German J Clin Oncol, 2007, 6(1):27-31.
25. Bartke A, Westbrook R, Sun L, et al. Links between growth hormone and aging. Endokrynol Pol, 2013, 64(1):46-52.

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CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of Small Interfering RNA Targeting Inhibition of Growth Hormone Receptor on Proliferation and Invasion of Human Liver Cancer Line SMMC-7721

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