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Published in: Pathology & Oncology Research 1/2018

01-01-2018 | Original Article

RACK1 Silencing Induces Cell Apoptosis and Inhibits Cell Proliferation in Hepatocellular Carcinoma MHCC97-H Cells

Authors: Yuan-hang Zou, Xue-dong Li, Qi-hao Zhang, De-zhong Liu

Published in: Pathology & Oncology Research | Issue 1/2018

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Abstract

This study aimed to explore the effects of RACK1 gene silencing on the apoptosis and proliferation of hepatocellular carcinoma (HCC) MHCC97-H cells. After transfecting MHCC97-H cells with siRNA, RACK1 gene silencing model was established. The cells were divided into blank group, siRNA group and empty plasmid group, respectively. The mRNA and protein expressions of RACK1, cyclin D1 and BAX were determined by qRT-PCR and Western blotting. CCK-8 assay, flow cytometry and FITC-Annexin V/PI staining were used to determine cell viability, cell cycle and cell apoptosis, respectively. The results of qRT-PCR and Western blotting suggested that when compared with the blank group and the empty plasmid group, the mRNA and protein expressions of RACK1 and Cyclin D1 decreased significantly while the mRNA and protein BAX expressions increased substantially in the siRNA group (all P < 0.05). The results of CCK-8 assay revealed that the siRNA group exhibited significantly lower cell viability when compared with the blank group and the empty plasmid group (both P < 0.05); and the cell viability in the siRNA group decreased gradually with the increase of time. The results of flow cytometry and FITC-Annexin V/PI staining indicated that when compared with the blank group and the empty plasmid group, the proportion of cells in S phase was markedly lower and the apoptosis rate was significantly higher in the siRNA group (both P < 0.05). Our study suggests that inhibition of RACK1 could suppress cell proliferation and induce apoptosis in HCC MHCC97-H cells.
Literature
1.
go back to reference Fonseca AL, Cha CH (2014) Hepatocellular carcinoma: a comprehensive overview of surgical therapy. J Surg Oncol 110(6):712–719CrossRefPubMed Fonseca AL, Cha CH (2014) Hepatocellular carcinoma: a comprehensive overview of surgical therapy. J Surg Oncol 110(6):712–719CrossRefPubMed
2.
go back to reference Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65(2):87–108CrossRefPubMed Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65(2):87–108CrossRefPubMed
4.
go back to reference El-Serag HB, Rudolph KL (2007) Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 132(7):2557–2576CrossRefPubMed El-Serag HB, Rudolph KL (2007) Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 132(7):2557–2576CrossRefPubMed
5.
go back to reference Schlesinger S, Aleksandrova K, Pischon T, Fedirko V, Jenab M, Trepo E, Boffetta P, Dahm CC, Overvad K, Tjonneland A, Halkjaer J, Fagherazzi G, Boutron-Ruault MC, Carbonnel F, Kaaks R, Lukanova A, Boeing H, Trichopoulou A, Bamia C, Lagiou P, Palli D, Grioni S, Panico S, Tumino R, Vineis P, Bueno-de-Mesquita HB, van den Berg S, Peeters PH, Braaten T, Weiderpass E, Quiros JR, Travier N, Sanchez MJ, Navarro C, Barricarte A, Dorronsoro M, Lindkvist B, Regner S, Werner M, Sund M, Khaw KT, Wareham N, Travis RC, Norat T, Wark PA, Riboli E, Nothlings U (2013) Abdominal obesity, weight gain during adulthood and risk of liver and biliary tract cancer in a European cohort. Int J Cancer 132(3):645–657CrossRefPubMed Schlesinger S, Aleksandrova K, Pischon T, Fedirko V, Jenab M, Trepo E, Boffetta P, Dahm CC, Overvad K, Tjonneland A, Halkjaer J, Fagherazzi G, Boutron-Ruault MC, Carbonnel F, Kaaks R, Lukanova A, Boeing H, Trichopoulou A, Bamia C, Lagiou P, Palli D, Grioni S, Panico S, Tumino R, Vineis P, Bueno-de-Mesquita HB, van den Berg S, Peeters PH, Braaten T, Weiderpass E, Quiros JR, Travier N, Sanchez MJ, Navarro C, Barricarte A, Dorronsoro M, Lindkvist B, Regner S, Werner M, Sund M, Khaw KT, Wareham N, Travis RC, Norat T, Wark PA, Riboli E, Nothlings U (2013) Abdominal obesity, weight gain during adulthood and risk of liver and biliary tract cancer in a European cohort. Int J Cancer 132(3):645–657CrossRefPubMed
6.
go back to reference Maluccio M, Covey A (2012) Recent progress in understanding, diagnosing, and treating hepatocellular carcinoma. CA Cancer J Clin 62(6):394–399CrossRefPubMed Maluccio M, Covey A (2012) Recent progress in understanding, diagnosing, and treating hepatocellular carcinoma. CA Cancer J Clin 62(6):394–399CrossRefPubMed
7.
go back to reference Gluer AM, Cocco N, Laurence JM, Johnston ES, Hollands MJ, Pleass HC, Richardson AJ, Lam VW (2012) Systematic review of actual 10-year survival following resection for hepatocellular carcinoma. HPB (Oxford) 14(5):285–290CrossRef Gluer AM, Cocco N, Laurence JM, Johnston ES, Hollands MJ, Pleass HC, Richardson AJ, Lam VW (2012) Systematic review of actual 10-year survival following resection for hepatocellular carcinoma. HPB (Oxford) 14(5):285–290CrossRef
8.
go back to reference Wu J, Meng J, Du Y, Huang Y, Jin Y, Zhang J, Wang B, Zhang Y, Sun M, Tang J (2013) RACK1 promotes the proliferation, migration and invasion capacity of mouse hepatocellular carcinoma cell line in vitro probably by PI3K/Rac1 signaling pathway. Biomed Pharmacother 67(4):313–319CrossRefPubMed Wu J, Meng J, Du Y, Huang Y, Jin Y, Zhang J, Wang B, Zhang Y, Sun M, Tang J (2013) RACK1 promotes the proliferation, migration and invasion capacity of mouse hepatocellular carcinoma cell line in vitro probably by PI3K/Rac1 signaling pathway. Biomed Pharmacother 67(4):313–319CrossRefPubMed
9.
go back to reference Wang WD, Wen Z, Ji W, Ma Y (2015) RACK1 expression contributes to JNK activity, but JNK activity does not enhance RACK1 expression in hepatocellular carcinoma SMMC-7721 cells. Oncol Lett 9(6):2767–2770PubMedPubMedCentral Wang WD, Wen Z, Ji W, Ma Y (2015) RACK1 expression contributes to JNK activity, but JNK activity does not enhance RACK1 expression in hepatocellular carcinoma SMMC-7721 cells. Oncol Lett 9(6):2767–2770PubMedPubMedCentral
10.
go back to reference He X, Wang J, Messing EM, Wu G (2011) Regulation of receptor for activated C kinase 1 protein by the von Hippel-Lindau tumor suppressor in IGF-I-induced renal carcinoma cell invasiveness. Oncogene 30(5):535–547CrossRefPubMed He X, Wang J, Messing EM, Wu G (2011) Regulation of receptor for activated C kinase 1 protein by the von Hippel-Lindau tumor suppressor in IGF-I-induced renal carcinoma cell invasiveness. Oncogene 30(5):535–547CrossRefPubMed
12.
13.
go back to reference Lu F, Zhang C, Wu WJ, Wu YM (2012) RACK1 downregulation suppresses migration and proliferation of neuroblastoma cell lines. Oncol Rep 27(5):646–1652 Lu F, Zhang C, Wu WJ, Wu YM (2012) RACK1 downregulation suppresses migration and proliferation of neuroblastoma cell lines. Oncol Rep 27(5):646–1652
14.
go back to reference Li J, Guo Y, Feng X, Wang Z, Wang Y, Deng P, Zhang D, Wang R, Xie L, Xu X, Zhou Y, Ji N, Hu J, Zhou M, Liao G, Geng N, Jiang L, Wang Z, Chen Q (2012) Receptor for activated C kinase 1 (RACK1): a regulator for migration and invasion in oral squamous cell carcinoma cells. J Cancer Res Clin Oncol 138(4):563–571CrossRefPubMed Li J, Guo Y, Feng X, Wang Z, Wang Y, Deng P, Zhang D, Wang R, Xie L, Xu X, Zhou Y, Ji N, Hu J, Zhou M, Liao G, Geng N, Jiang L, Wang Z, Chen Q (2012) Receptor for activated C kinase 1 (RACK1): a regulator for migration and invasion in oral squamous cell carcinoma cells. J Cancer Res Clin Oncol 138(4):563–571CrossRefPubMed
15.
go back to reference Guo Y, Wang W, Wang J, Feng J, Wang Q, Jin J, Lv M, Li X, Li Y, Ma Y, Shen B, Zhang J (2013) Receptor for activated C kinase 1 promotes hepatocellular carcinoma growth by enhancing mitogen-activated protein kinase kinase 7 activity. Hepatology 57:140–151CrossRefPubMed Guo Y, Wang W, Wang J, Feng J, Wang Q, Jin J, Lv M, Li X, Li Y, Ma Y, Shen B, Zhang J (2013) Receptor for activated C kinase 1 promotes hepatocellular carcinoma growth by enhancing mitogen-activated protein kinase kinase 7 activity. Hepatology 57:140–151CrossRefPubMed
16.
go back to reference McCahill A, Warwicker J, Bolger GB, Houslay MD, Yarwood SJ (2002) The RACK1 scaffold protein: a dynamic cog in cell response mechanisms. Mol Pharmacol 62(6):1261–1273CrossRefPubMed McCahill A, Warwicker J, Bolger GB, Houslay MD, Yarwood SJ (2002) The RACK1 scaffold protein: a dynamic cog in cell response mechanisms. Mol Pharmacol 62(6):1261–1273CrossRefPubMed
17.
go back to reference Wang F, Osawa T, Tsuchida R, Yuasa Y, Shibuya M (2011) Downregulation of receptor for activated C-kinase 1 (RACK1) suppresses tumor growth by inhibiting tumor cell proliferation and tumor-associated angiogenesis. Cancer Sci 102(11):2007–2013CrossRefPubMed Wang F, Osawa T, Tsuchida R, Yuasa Y, Shibuya M (2011) Downregulation of receptor for activated C-kinase 1 (RACK1) suppresses tumor growth by inhibiting tumor cell proliferation and tumor-associated angiogenesis. Cancer Sci 102(11):2007–2013CrossRefPubMed
18.
go back to reference Rajput S, Kumar BN, Dey KK, Pal I, Parekh A, Mandal M (2013) Molecular targeting of Akt by thymoquinone promotes G(1) arrest through translation inhibition of cyclin D1 and induces apoptosis in breast cancer cells. Life Sci 93(21):783–790CrossRefPubMed Rajput S, Kumar BN, Dey KK, Pal I, Parekh A, Mandal M (2013) Molecular targeting of Akt by thymoquinone promotes G(1) arrest through translation inhibition of cyclin D1 and induces apoptosis in breast cancer cells. Life Sci 93(21):783–790CrossRefPubMed
19.
go back to reference Peng R, Jiang B, Ma J, Ma Z, Wan X, Liu H, Chen Z, Cheng Q, Chen R (2013) Forced downregulation of RACK1 inhibits glioma development by suppressing Src/Akt signaling activity. Oncol Rep 30(5):2195–2202CrossRefPubMed Peng R, Jiang B, Ma J, Ma Z, Wan X, Liu H, Chen Z, Cheng Q, Chen R (2013) Forced downregulation of RACK1 inhibits glioma development by suppressing Src/Akt signaling activity. Oncol Rep 30(5):2195–2202CrossRefPubMed
20.
go back to reference Mamidipudi V, Miller LD, Mochly-Rosen D, Cartwright CA (2007) Peptide modulators of Src activity in G1 regulate entry into S phase and proliferation of NIH 3T3 cells. Biochem Biophys Res Commun 352(2):423–430CrossRefPubMed Mamidipudi V, Miller LD, Mochly-Rosen D, Cartwright CA (2007) Peptide modulators of Src activity in G1 regulate entry into S phase and proliferation of NIH 3T3 cells. Biochem Biophys Res Commun 352(2):423–430CrossRefPubMed
21.
go back to reference Lopez-Bergami P, Huang C, Goydos JS, Yip D, Bar-Eli M, Herlyn M, Smalley KS, Mahale A, Eroshkin A, Aaronson S, Ronai Z (2007) Rewired ERK-JNK signaling pathways in melanoma. Cancer Cell 11(5):447–460CrossRefPubMedPubMedCentral Lopez-Bergami P, Huang C, Goydos JS, Yip D, Bar-Eli M, Herlyn M, Smalley KS, Mahale A, Eroshkin A, Aaronson S, Ronai Z (2007) Rewired ERK-JNK signaling pathways in melanoma. Cancer Cell 11(5):447–460CrossRefPubMedPubMedCentral
22.
go back to reference (2015) Inhibition of Notch- and EGFR signaling reduces cell viability and angiogenesis in glioblastoma multiforme. Journal/Cancer Research 75: 1369–1369. (2015) Inhibition of Notch- and EGFR signaling reduces cell viability and angiogenesis in glioblastoma multiforme. Journal/Cancer Research 75: 1369–1369.
23.
go back to reference Sutton P, Borgia JA, Bonomi P, Plate JM (2013) Lyn, a Src family kinase, regulates activation of epidermal growth factor receptors in lung adenocarcinoma cells. Mol Cancer 12:76CrossRefPubMedPubMedCentral Sutton P, Borgia JA, Bonomi P, Plate JM (2013) Lyn, a Src family kinase, regulates activation of epidermal growth factor receptors in lung adenocarcinoma cells. Mol Cancer 12:76CrossRefPubMedPubMedCentral
24.
go back to reference Wilson JM, Kunnimalaiyaan S, Kunnimalaiyaan M, Gamblin TC (2015) Inhibition of the AKT pathway in cholangiocarcinoma by MK2206 reduces cellular viability via induction of apoptosis. Cancer Cell Int 15(1):13CrossRefPubMedPubMedCentral Wilson JM, Kunnimalaiyaan S, Kunnimalaiyaan M, Gamblin TC (2015) Inhibition of the AKT pathway in cholangiocarcinoma by MK2206 reduces cellular viability via induction of apoptosis. Cancer Cell Int 15(1):13CrossRefPubMedPubMedCentral
Metadata
Title
RACK1 Silencing Induces Cell Apoptosis and Inhibits Cell Proliferation in Hepatocellular Carcinoma MHCC97-H Cells
Authors
Yuan-hang Zou
Xue-dong Li
Qi-hao Zhang
De-zhong Liu
Publication date
01-01-2018
Publisher
Springer Netherlands
Published in
Pathology & Oncology Research / Issue 1/2018
Print ISSN: 1219-4956
Electronic ISSN: 1532-2807
DOI
https://doi.org/10.1007/s12253-017-0214-6

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