miR-184通过JNK信号通路调控卵巢癌的增殖与凋亡
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摘要
目的:探讨miR-184通过JNK信号通路对卵巢癌增殖与凋亡的影响。方法:收集卵巢癌患者的癌组织与癌旁组织标本,荧光定量PCR检测miR-184表达;采用Lipofectamine 2000将miR-184 inhibitor和miR-184 mimic分别转入卵巢癌细胞株OVCAR3,qPCR检测其转染效率,MTT及细胞凋亡实验检测miR-184表达对细胞增殖及凋亡的影响;Westerm blot检测miR-184表达对JNK信号通路中total JNK及p-JNK蛋白的影响。结果:卵巢癌组织中miR-184表达较癌旁组织明显升高,差异有统计学意义(P<0.05);高表达miR-184后,OVCAR3细胞增殖率显著提高,细胞凋亡率显著下降,磷酸化p-JNK表达明显上调(P均<0.05);下调miR-184表达后,OVCAR3细胞增殖率明显下降,细胞凋亡率显著升高,磷酸化p-JNK表达明显下调(P均<0.05)。结论:miR-184可促进卵巢癌细胞增殖并抑制细胞凋亡,其机制可能与JNK信号通路有关。
Objective:To investigate the effect of microRNA-184 on proliferation and apoptosis of ovarian cancer through JNK signaling pathway.Methods:The expression of microRNA-184 in ovarian cancer tissues and adjacent tissues was detected by fluorescent quantitative PCR,and the expression of microRNA-184 inhibitor and microRNA-184 mimic were transfected into ovarian cancer cell line OVCAR3 by Lipofectamine 2000.The transfection efficiency was detected by qPCR,and the effects of microRNA-184 expression on cell proliferation and apoptosis were detected by MTT and cell apoptosis experiments.The expression of microRNA-184 was detected by Western blot and it's effects of alteration on total JNK and p-JNK proteins in JNK signaling pathway.Results:The expression of microRNA-184 in ovarian cancer tissues was significantly higher than that in adjacent tissues(P<0.05).After high microRNA-184,the proliferation rate and apoptotic rate of OVCAR3 cells were significantly increased,and the expression of phosphorylated p-JNK in OVCAR3 cells was significantly increased(P<0.05),while after down-regulation of microRNA-184,the proliferation rate and apoptotic rate of OVCAR3 cells were significantly decreased.The expression of phosphorylated p-JNK was significantly down-regulated(P<0.05).Conclusions:MiR-184 can promote the proliferation and inhibit the apoptosis of ovarian cancer cells,and its mechanism may be related to JNK signaling pathway.
Objective:To investigate the effect of microRNA-184 on proliferation and apoptosis of ovarian cancer through JNK signaling pathway.Methods:The expression of microRNA-184 in ovarian cancer tissues and adjacent tissues was detected by fluorescent quantitative PCR,and the expression of microRNA-184 inhibitor and microRNA-184 mimic were transfected into ovarian cancer cell line OVCAR3 by Lipofectamine 2000.The transfection efficiency was detected by qPCR,and the effects of microRNA-184 expression on cell proliferation and apoptosis were detected by MTT and cell apoptosis experiments.The expression of microRNA-184 was detected by Western blot and it's effects of alteration on total JNK and p-JNK proteins in JNK signaling pathway.Results:The expression of microRNA-184 in ovarian cancer tissues was significantly higher than that in adjacent tissues(P<0.05).After high microRNA-184,the proliferation rate and apoptotic rate of OVCAR3 cells were significantly increased,and the expression of phosphorylated p-JNK in OVCAR3 cells was significantly increased(P<0.05),while after down-regulation of microRNA-184,the proliferation rate and apoptotic rate of OVCAR3 cells were significantly decreased.The expression of phosphorylated p-JNK was significantly down-regulated(P<0.05).Conclusions:MiR-184 can promote the proliferation and inhibit the apoptosis of ovarian cancer cells,and its mechanism may be related to JNK signaling pathway.
引文
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[3] 李泽夏,刘海玲,吕辉,等.与卵巢癌转移有关的microRNA研究进展[J].现代生物医学进展,2011,11(14):2793-2796
[4] 朱清,许波,刘雨生.microRNA在卵巢和卵巢癌中的表达及功能[J].安徽医科大学学报,2014,49(5):694-697
[5] Lee RC,Feinbaum RL,Ambros V.The C.elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 [J].Cell,1993,75(5):843-854
[6] Iorio MV,Visone R,Di Leva G,et al.MicroRNA signatures in human ovarian cancer [J].Cancer Res,2007,67(18):8699-8707
[7] 冷慧敏,钱卫平,周亮,等.肾癌中MIR-184的异常表达及意义[J].北京大学学报(医学版),2011,43(4):509-513
[8] 郎博娟,马金阳,胡余昌,等.胶质瘤中MicroRNA-184的表达及其预后价值[J].肿瘤防治研究,2015,42(5):483-487
[9] 邹丰,王绪明,刘丽江.ER-α36介导的胃癌中miRNA表达的研究[J].世界华人消化杂志,2013,24:2373-2377
[10] 张爱臣,佟玲玲.卵巢癌的诊治进展[J].中国计划生育和妇产科,2013,5(1):5-8
[11] 李楠,张梦真.MicroRNA在卵巢癌中的研究进展[J].现代妇产科进展,2013,22(9):765-766
[12] Dweep H,Sticht C,Pandey P,et al.miRWalk-database:prediction of possible miRNA binding sites by “walking” the genes of three genomes[J].J Biomed Informat,2011,44(5):839-847
[13] Kong D,Liu Y,Liu Q,et al.The retinal determination gene network:from developmental regulator to cancer therapeutic target[J].Oncotarget,2016,7(31):50755-50765
[14] 陶陶,王敏.化疗耐药及敏感卵巢癌细胞差异表达microRNA的筛查与组织鉴定[J].中国医科大学学报,2013,42(2):118-122
[15] Nam EJ,Yoon H,Kim SW,et al.MicroRNA expression profiles in serous ovarian carcinoma [J].Clin Cancer Res,2008,14(9):2690-2695
[16] 双婷,吴建磊,朱彦丽.耐药与敏感卵巢癌组织中microRNA 表达差异谱检测及分析[J].中国实用妇科与产科杂志,2013,29(1):33-35
[17] 赖永旭,李艳萍.MicroRNA-184靶向作用EPB41L5抑制肾细胞癌存活与迁移研究[J].陕西医学杂志,2017,46(3):294-297
[18] 刘洋,周红林,侯友芳,等.miR-214负向调控卵巢癌细胞株SKOV-3中Sema4D的表达[J].中国妇幼保健,2015,30(3):431-433
[19] 张旗,何湘君,马丽萍,等.p53通路微小RNA在卵巢癌细胞和浆液性卵巢癌组织中的表达及意义[J].中华肿瘤杂志,2011,33(12):885-890
[20] Zhang C,Tian W,Meng L,et al.PRL-3 promotes gastric cancer migration and invasion through a NF-κB-HIF-1α-miR-210 axis[J].J Mol Med,2016,94(4):401-415
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