RNAi抑制NOB1表达对肝癌BEL-7402细胞增殖及p38MAPK磷酸化水平的影响
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摘要
目的:探讨RNAi下调Nin1结合蛋白(NOB1)表达对肝癌细胞增殖及p38丝裂原活化蛋白激酶(p38MAPK)磷酸化水平的影响。方法:以肝癌BEL-7402细胞作为研究对象,将肝癌细胞分为空白对照组、siRNANC组、NOB1 siRNA组3组,其中siRNA-NC组、NOB1 siRNA组分别为稳定感染阴性对照慢病毒和NOB1 siRNA慢病毒的肝癌细胞,空白对照组为不做慢病毒感染的肝癌细胞。用qRT-PCR和Western blot检测干扰效果。CCK-8法测定细胞增殖情况,流式细胞仪检测细胞凋亡情况,Western blot法检测细胞中磷酸化p38MAPK和活化的Caspase-3蛋白的表达情况。结果:NOB1 siRNA组细胞中NOB1表达水平、细胞增殖活性降低,细胞凋亡率升高,磷酸化p38MAPK和活化的Caspase-3蛋白表达水平升高,与空白对照组、siRNA-NC组比较,差异均有统计学意义(P <0. 05)。结论:下调NOB1表达具有抑制BEL-7402细胞增殖、诱导细胞凋亡、促进细胞中p38MAPK磷酸化的作用。
Aim: To investigate the effects of down-regulation of Nin one binding protein( NOB1) expression by RNAi on the proliferation and phosphorylation of p38 MAPK in hepatocellular carcinoma cells. Methods: Hepatocellular carcinoma BEL-7402 cells were divided into three groups: blank control group,siRNA-NC group and NOB1 siRNA group. In siRNA-NC group and NOB1 siRNA group,BEL-7402 cells were infected with negative control lentivirus and NOB1 siRNA lentivirus respectively. The blank control group was cells without lentivirus infection. The interference effects were detected by qRT-PCR and Western blot. Cell proliferation activity was measured by CCK-8 assay. Flow cytometry was used to detect apoptosis. Western blot was used to detect the level of phosphorylated p38 MAPK and activated Caspase-3 proteins in cells.Results: Compared with blank control group and siRNA-NC group,in NOB1 siRNA group,the expression of NOB1 and cell proliferation activity were decreased,apoptotic rate was increased,and the phosphorylated p38 MAPK and activated Caspase-3 protein levels were increased( P < 0. 05). Conclusion: Down-regulation of NOB1 expression could inhibit the proliferation of BEL-7402 cells,induce apoptosis,and promote the phosphorylation of p38 MAPK.
Aim: To investigate the effects of down-regulation of Nin one binding protein( NOB1) expression by RNAi on the proliferation and phosphorylation of p38 MAPK in hepatocellular carcinoma cells. Methods: Hepatocellular carcinoma BEL-7402 cells were divided into three groups: blank control group,siRNA-NC group and NOB1 siRNA group. In siRNA-NC group and NOB1 siRNA group,BEL-7402 cells were infected with negative control lentivirus and NOB1 siRNA lentivirus respectively. The blank control group was cells without lentivirus infection. The interference effects were detected by qRT-PCR and Western blot. Cell proliferation activity was measured by CCK-8 assay. Flow cytometry was used to detect apoptosis. Western blot was used to detect the level of phosphorylated p38 MAPK and activated Caspase-3 proteins in cells.Results: Compared with blank control group and siRNA-NC group,in NOB1 siRNA group,the expression of NOB1 and cell proliferation activity were decreased,apoptotic rate was increased,and the phosphorylated p38 MAPK and activated Caspase-3 protein levels were increased( P < 0. 05). Conclusion: Down-regulation of NOB1 expression could inhibit the proliferation of BEL-7402 cells,induce apoptosis,and promote the phosphorylation of p38 MAPK.
引文
[1]RAVAIOLI M,GRAZI GL,ERCOLANI G,et al.Liver allocation for hepatocellular carcinoma:a European center policy in the pre-MELD era[J].Transplantation,2006,81(4):525
[2]JAGANATHAN A,MURUGAN K,PANNEERSELVAM C,et al.Earthworm-mediated synthesis of silver nanoparticles:a potent tool against hepatocellular carcinoma,plasmodium falciparum parasites and malaria mosquitoes[J].Parasitol Int,2016,65(3):276
[3]DAI H,HOU K,CAI Z,et al.Low-level miR-646 in colorectal cancer inhibits cell proliferation and migration by targeting NOB1 expression[J].Oncol Lett,2017,14(6):6708
[4]WANG J,CAO L,WU J,et al.Long non-coding RNASNHG1 regulates NOB1 expression by sponging miR-326and promotes tumorigenesis in osteosarcoma[J].Int J Oncol,2018,52(1):77
[5]WANG H,LI P,ZHAO B.Knockdown of NOB1 expression by RNAi inhibits cellular proliferation and migration in human gliomas[J].Gene,2013,528(2):146
[6]张媛媛,宋育林.NOB1在胃癌组织中的表达及临床意义[J].安徽医药,2013,17(10):1707
[7]赵丽娟,李志锋,石荣亚,等.NOB1在肝癌组织中的表达及意义[J].临床肝胆病杂志,2016,32(4):739
[8]KONG RR,LIU W,GUO YR,et al.Inhibition of NOB1 by microRNA-330-5p overexpression represses cell growth of nonsmall cell lung cancer[J].Oncol Rep,2017,38(4):2572
[9]LEE JC,TSENG CK,YOUNG KC,et al.Andrographolide exerts anti-hepatitis C virus activity by up-regulating haeme oxygenase-1 via the p38MAPK/Nrf2 pathway in human hepatoma cells[J].Br J Pharmacol,2014,171(1):237
[10]MENG W,WANG PS,LIU J,et al.Adenovirus-mediated siRNA targeting NOB1 inhibits tumor growth and enhances radiosensitivity of human papillary thyroid carcinoma in vitro and in vivo[J].Oncol Rep,2014,32(6):2411
[11]LUO LM,WANG YH,YIN YR,et al.Effects of NOB1 on the pathogenesis of osteosarcoma and its expression on the chemosensitivity to cisplatin[J].Oncol Lett,2018,15(3):3548
[12]LIN Y,PENG S,YU H,et al.RNAi-mediated downregulation of NOB1 suppresses the growth and colony-formation ability of human ovarian cancer cells[J].Med Oncol,2012,29(1):311
[13]LIN Y,XU TM,ZHOU SQ,et al.MicroRNA-363 inhibits ovarian cancer progression by inhibiting NOB1[J].Oncotarget,2017,8(60):101649
[14]张玉洁,王勉,周威,等.NOB1在食管鳞状细胞癌中的表达及意义[J].现代肿瘤医学,2013,21(4):756
[15]LOPERGOLO A,PENNATI M,GANDELLINI P,et al.Apollon gene silencing induces apoptosis in breast cancer cells through p53 stabilisation and caspase-3 activation[J].Br J Cancer,2009,100(5):739
[16]CHECINSKA A,HOOGELAND BS,RODRIGUEZ JA,et al.Role of XIAP in inhibiting cisplatin-induced caspase activation in non-small cell lung cancer cells:a small molecule Smac mimic sensitizes for chemotherapy-induced apoptosis by enhancing caspase-3 activation[J].Exp Cell Res,2007,313(6):1215
[17]吴艳林,万里新,屈中玉,等.Noblp人类同源基因在食管癌组织中的表达及对食管癌细胞凋亡的影响[J].中华实验外科杂志,2017,34(6):1066
[18]黎雁英,田佳,陈石涛,等.NOB1反义寡核苷酸对人卵巢癌细胞生长、凋亡的影响[J].安徽医科大学学报,2013,48(10):1174
[19]张奇,白晓东,付小兵.p38MAPK信号通路研究进展[J].感染、炎症、修复,2005,6(2):121
[20]HE H,CHANG R,ZHANG T,et al.ATM mediates DAB2IP-deficient bladder cancer cell resistance to ionizing radiation through the p38MAPK and NF-κB signaling pathway[J].Mol Med Rep,2017,16(2):1216
[2]JAGANATHAN A,MURUGAN K,PANNEERSELVAM C,et al.Earthworm-mediated synthesis of silver nanoparticles:a potent tool against hepatocellular carcinoma,plasmodium falciparum parasites and malaria mosquitoes[J].Parasitol Int,2016,65(3):276
[3]DAI H,HOU K,CAI Z,et al.Low-level miR-646 in colorectal cancer inhibits cell proliferation and migration by targeting NOB1 expression[J].Oncol Lett,2017,14(6):6708
[4]WANG J,CAO L,WU J,et al.Long non-coding RNASNHG1 regulates NOB1 expression by sponging miR-326and promotes tumorigenesis in osteosarcoma[J].Int J Oncol,2018,52(1):77
[5]WANG H,LI P,ZHAO B.Knockdown of NOB1 expression by RNAi inhibits cellular proliferation and migration in human gliomas[J].Gene,2013,528(2):146
[6]张媛媛,宋育林.NOB1在胃癌组织中的表达及临床意义[J].安徽医药,2013,17(10):1707
[7]赵丽娟,李志锋,石荣亚,等.NOB1在肝癌组织中的表达及意义[J].临床肝胆病杂志,2016,32(4):739
[8]KONG RR,LIU W,GUO YR,et al.Inhibition of NOB1 by microRNA-330-5p overexpression represses cell growth of nonsmall cell lung cancer[J].Oncol Rep,2017,38(4):2572
[9]LEE JC,TSENG CK,YOUNG KC,et al.Andrographolide exerts anti-hepatitis C virus activity by up-regulating haeme oxygenase-1 via the p38MAPK/Nrf2 pathway in human hepatoma cells[J].Br J Pharmacol,2014,171(1):237
[10]MENG W,WANG PS,LIU J,et al.Adenovirus-mediated siRNA targeting NOB1 inhibits tumor growth and enhances radiosensitivity of human papillary thyroid carcinoma in vitro and in vivo[J].Oncol Rep,2014,32(6):2411
[11]LUO LM,WANG YH,YIN YR,et al.Effects of NOB1 on the pathogenesis of osteosarcoma and its expression on the chemosensitivity to cisplatin[J].Oncol Lett,2018,15(3):3548
[12]LIN Y,PENG S,YU H,et al.RNAi-mediated downregulation of NOB1 suppresses the growth and colony-formation ability of human ovarian cancer cells[J].Med Oncol,2012,29(1):311
[13]LIN Y,XU TM,ZHOU SQ,et al.MicroRNA-363 inhibits ovarian cancer progression by inhibiting NOB1[J].Oncotarget,2017,8(60):101649
[14]张玉洁,王勉,周威,等.NOB1在食管鳞状细胞癌中的表达及意义[J].现代肿瘤医学,2013,21(4):756
[15]LOPERGOLO A,PENNATI M,GANDELLINI P,et al.Apollon gene silencing induces apoptosis in breast cancer cells through p53 stabilisation and caspase-3 activation[J].Br J Cancer,2009,100(5):739
[16]CHECINSKA A,HOOGELAND BS,RODRIGUEZ JA,et al.Role of XIAP in inhibiting cisplatin-induced caspase activation in non-small cell lung cancer cells:a small molecule Smac mimic sensitizes for chemotherapy-induced apoptosis by enhancing caspase-3 activation[J].Exp Cell Res,2007,313(6):1215
[17]吴艳林,万里新,屈中玉,等.Noblp人类同源基因在食管癌组织中的表达及对食管癌细胞凋亡的影响[J].中华实验外科杂志,2017,34(6):1066
[18]黎雁英,田佳,陈石涛,等.NOB1反义寡核苷酸对人卵巢癌细胞生长、凋亡的影响[J].安徽医科大学学报,2013,48(10):1174
[19]张奇,白晓东,付小兵.p38MAPK信号通路研究进展[J].感染、炎症、修复,2005,6(2):121
[20]HE H,CHANG R,ZHANG T,et al.ATM mediates DAB2IP-deficient bladder cancer cell resistance to ionizing radiation through the p38MAPK and NF-κB signaling pathway[J].Mol Med Rep,2017,16(2):1216
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