微小核糖核酸-34a调节高迁移率族蛋白B1对卵巢癌细胞增殖的影响
|
摘要
目的探讨微小核糖核酸-34a(miR-34a)通过调节高迁移率族蛋白B1(HMGB1)对卵巢癌SKOV-3细胞增殖能力的影响,了解miR-34a参与卵巢癌发生、发展的分子机制。方法设计miR-34a模拟物、miR-34a抑制剂,对SKOV-3细胞进行转染,采用CCK8检测转染24 h、48 h和72 h后SKOV-3细胞存活情况,采用qRT-PCR法检测各组miR-34a和HMGB1 mRNA的表达;利用Western blot法检测各组HMGB1蛋白的表达。结果转染miR-34a模拟物后,SKOV-3细胞的增殖率显著低于空白对照组和miR-34a模拟物对照组(P均<0.05),HMGB1 mRNA和蛋白表达量较空白对照组和miR-34a模拟物对照组降低(P均<0.05);转染miR-34a抑制剂后,SKOV-3细胞的增殖率显著高于空白对照组和miR-34a抑制剂对照组(P均<0.05),HMGB1 mRNA和蛋白表达量较空白对照组和miR-34a抑制剂对照组增加(P均<0.05)。结论 MiR-34a影响卵巢癌细胞的增殖功能,其可能是通过抑制HMGB1的表达参与卵巢癌的发生、发展。
Objective To investigate the inhibitory effect of microRNA-34a(miR-34a) on ovarian cancer SKOV-3 cells proliferation by regulating high mobility group box 1(HMGB1).Methods Chemically synthesized miR-34a mimics, miR-34a inhibitors were designed, and were transfected into SKOV-3 cells. The ability of proliferation were detected by CCK8. The expressions of miR-34a and HMGB1 mRNA were detected by qRT-PCR. The expression of HMGB1 protein was detected by Western blot.Results After transfection with miR-34a mimics, the proliferative rate of SKOV3 cells was lower than those in the negative control group and the miR-34a mimics control group(P<0.05); the expression of HMGB1 was significantly down-regulated compared with the negative control group and the miR-34a mimics control group(P<0.05). However, after transfection with miR-34a inhibitors, the proliferative rate of SKOV3 cells was higher than those in the negative control group and the miR-34a inhibitors control group(P<0.05); the expression of HMGB1 was significantly up-regulated compared with the negative control group and the miR-34a inhibitors control group(P<0.05).Conclusion MiR-34a may inhibits the proliferation of ovarian cancer cells by regulating HMGB1.
Objective To investigate the inhibitory effect of microRNA-34a(miR-34a) on ovarian cancer SKOV-3 cells proliferation by regulating high mobility group box 1(HMGB1).Methods Chemically synthesized miR-34a mimics, miR-34a inhibitors were designed, and were transfected into SKOV-3 cells. The ability of proliferation were detected by CCK8. The expressions of miR-34a and HMGB1 mRNA were detected by qRT-PCR. The expression of HMGB1 protein was detected by Western blot.Results After transfection with miR-34a mimics, the proliferative rate of SKOV3 cells was lower than those in the negative control group and the miR-34a mimics control group(P<0.05); the expression of HMGB1 was significantly down-regulated compared with the negative control group and the miR-34a mimics control group(P<0.05). However, after transfection with miR-34a inhibitors, the proliferative rate of SKOV3 cells was higher than those in the negative control group and the miR-34a inhibitors control group(P<0.05); the expression of HMGB1 was significantly up-regulated compared with the negative control group and the miR-34a inhibitors control group(P<0.05).Conclusion MiR-34a may inhibits the proliferation of ovarian cancer cells by regulating HMGB1.
引文
[1] BROUWERVISSER J,LEE J,MCCULLAGH KA,et al.Insulin-like growth factor 2 silencing restores taxol sensitivity in drug resistant ovarian cancer[J].Plos One,2014,9(6):e100165.
[2] NAM EJ,YOON H,KIM SW,et al.MicroRNA expression profiles in serous ovarian carcinoma[J].Clin Cancer Res,2008,14(9):2690-2695.
[3] JELOVAC D,ARMSTRONG DK.Recent progress in the diagnosis and treatment of ovarian cancer[J].CA Cancer J Clin,2011,61(3):183-203.
[4] BARTEL DP.MicroRNAs:genomics,biogenesis,mechanism,and function[J].Cell,2004,116(2):281-297.
[5] 林兰,刘继斌.肝癌中CpG甲基化与microRNA-34沉默的研究[J].肿瘤基础与临床,2017,30(1):19-22.
[6] CHOI YS,LEE KE.The significance of miR-34a expression in endometrial carcinogenesis:correlation with expression of p16 and Ki-67 proteins in endometrial cancers[J].J Cancer Prev,2015,20(4):268-274.
[7] 王艳军,王高雄,黄天从,等.高迁移率族蛋白1通过调控Erk1/2、Cyclin D1及MMP14蛋白促进肝内胆管细胞癌进展[J].第三军医大学学报,2019,41(5):430-436.
[8] XIA J,YU X,SONG X,et al.Inhibiting the cytoplasmic location of HMGB1 reverses cisplatin resistance in human cervical cancer cells[J].Mol Med Rep,2017,15(1):488-494.
[9] ZHENG H,CHEN JN,YU X,et al.HMGB1 enhances drug resistance and promotes in vivo tumor growth of lung cancer cells[J].DNA Cell Biol,2016,35(10):622-627.
[10] YANG M,LIU L,XIE M,et al.Poly-ADP-ribosylation of HMGB1 regulates TNFSF10/TRAIL resistance through autophagy[J].Autophagy,2015,11(2):214-224.
[11] LIU K,HUANG J,XIE M,et al.MIR34A regulates autophagy and apoptosis by targeting HMGB1 in the retinoblastoma cell[J].Autophagy,2014,10(3):442-452.
[12] FARAZI TA,SPITZER JI,MOROZOV P,et al.miRNAs in human cancer[J].J Pathol,2011,223(2):102-115.
[13] O’CONNELL RM,RAO DS,CHAUDHURI AA,et al.Physiological and pathological roles for microRNAs in the immune system[J].Nat Rev Immunol,2010,10(2):111-122.
[14] VAN ROOIJ E,OLSON EN.MicroRNA therapeutics for cardiovascular disease:opportunities and obstacles[J].Nat Rev Drug Discov,2012,11(11):860-872.
[15] 成兵,吴信华.miR-34a调控Survivin表达对膀胱癌细胞生物学行为的影响[J].南通大学学报(医学版),2012,32(5):381-385.
[16] LANGE SS,MITCHELL DL,VASQUEZ KM.High mobility group protein B1 enhances DNA repair and chromatin modification after DNA damage[J].Proc Natl Acad Sci U S A,2008,105(30):10320-10325.
[17] APETOH L,GHIRINGHELLI F,TESNIERE A,et al.Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy[J].Nat Med,2007,13(9):1050-1059.
[18] TANG D,KANG R,CHEH CW,et al.HMGB1 release and redox regulates autophagy and apoptosis in cancer cells[J].Oncogene,2010,29(38):5299-5310.
[19] LUO Y,OHMORI H,FUJII K,et al.HMGB1 attenuates anti-metastatic defence of the liver in colorectal cancer[J].Eur J Cancer,2010,46(4):791-799.
[2] NAM EJ,YOON H,KIM SW,et al.MicroRNA expression profiles in serous ovarian carcinoma[J].Clin Cancer Res,2008,14(9):2690-2695.
[3] JELOVAC D,ARMSTRONG DK.Recent progress in the diagnosis and treatment of ovarian cancer[J].CA Cancer J Clin,2011,61(3):183-203.
[4] BARTEL DP.MicroRNAs:genomics,biogenesis,mechanism,and function[J].Cell,2004,116(2):281-297.
[5] 林兰,刘继斌.肝癌中CpG甲基化与microRNA-34沉默的研究[J].肿瘤基础与临床,2017,30(1):19-22.
[6] CHOI YS,LEE KE.The significance of miR-34a expression in endometrial carcinogenesis:correlation with expression of p16 and Ki-67 proteins in endometrial cancers[J].J Cancer Prev,2015,20(4):268-274.
[7] 王艳军,王高雄,黄天从,等.高迁移率族蛋白1通过调控Erk1/2、Cyclin D1及MMP14蛋白促进肝内胆管细胞癌进展[J].第三军医大学学报,2019,41(5):430-436.
[8] XIA J,YU X,SONG X,et al.Inhibiting the cytoplasmic location of HMGB1 reverses cisplatin resistance in human cervical cancer cells[J].Mol Med Rep,2017,15(1):488-494.
[9] ZHENG H,CHEN JN,YU X,et al.HMGB1 enhances drug resistance and promotes in vivo tumor growth of lung cancer cells[J].DNA Cell Biol,2016,35(10):622-627.
[10] YANG M,LIU L,XIE M,et al.Poly-ADP-ribosylation of HMGB1 regulates TNFSF10/TRAIL resistance through autophagy[J].Autophagy,2015,11(2):214-224.
[11] LIU K,HUANG J,XIE M,et al.MIR34A regulates autophagy and apoptosis by targeting HMGB1 in the retinoblastoma cell[J].Autophagy,2014,10(3):442-452.
[12] FARAZI TA,SPITZER JI,MOROZOV P,et al.miRNAs in human cancer[J].J Pathol,2011,223(2):102-115.
[13] O’CONNELL RM,RAO DS,CHAUDHURI AA,et al.Physiological and pathological roles for microRNAs in the immune system[J].Nat Rev Immunol,2010,10(2):111-122.
[14] VAN ROOIJ E,OLSON EN.MicroRNA therapeutics for cardiovascular disease:opportunities and obstacles[J].Nat Rev Drug Discov,2012,11(11):860-872.
[15] 成兵,吴信华.miR-34a调控Survivin表达对膀胱癌细胞生物学行为的影响[J].南通大学学报(医学版),2012,32(5):381-385.
[16] LANGE SS,MITCHELL DL,VASQUEZ KM.High mobility group protein B1 enhances DNA repair and chromatin modification after DNA damage[J].Proc Natl Acad Sci U S A,2008,105(30):10320-10325.
[17] APETOH L,GHIRINGHELLI F,TESNIERE A,et al.Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy[J].Nat Med,2007,13(9):1050-1059.
[18] TANG D,KANG R,CHEH CW,et al.HMGB1 release and redox regulates autophagy and apoptosis in cancer cells[J].Oncogene,2010,29(38):5299-5310.
[19] LUO Y,OHMORI H,FUJII K,et al.HMGB1 attenuates anti-metastatic defence of the liver in colorectal cancer[J].Eur J Cancer,2010,46(4):791-799.