miR-17-92基因簇增强前列腺癌DU145细胞的迁移、侵袭能力及对顺铂的耐药性

发布时间：2018-04-17 23:01

本文选题：miR-- + 前列腺肿瘤　；参考：《中国癌症杂志》2017年02期

【摘要】：背景与目的:mi R-17-92基因簇与多种疾病的发生密切相关,其在肺癌、肝癌、胃癌和前列腺癌等多种肿瘤细胞中均高表达。本研究利用慢病毒包装系统建立稳定高表达mi R-17-92基因簇的DU145细胞株,探讨mi R-17-92基因簇对前列腺癌DU145细胞的迁移、侵袭能力及对顺铂耐药性的影响。方法:构建高表达mi R-17-92基因簇的表达载体,转染DU145细胞株,同时转染空载体作为对照,并用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)进行鉴定。用x CELLigence系统监测细胞的迁移、侵袭能力及顺铂处理后的生长情况;通过划痕实验观察细胞的迁移情况;采用蛋白[质]印迹法(Western blot)、凝胶酶谱实验和RTFQ-PCR检测相关蛋白质和基因的表达以探讨mi R-17-92增强DU145细胞的迁移、侵袭能力及对顺铂耐药性的相关机制。结果:DU145-mi R-17-92细胞迁移速率和侵袭能力高于DU145-control细胞(P0.01)。DU145-mi R-17-92细胞中整合素β1的蛋白质表达水平和基质金属蛋白酶-9(matrix metalloprotein-9,MMP-9)的活性显著高于DU145-control细胞。顺铂处理后,DU145-mi R-17-92细胞的生长速度自12 h起快于DU145-control细胞并呈顺铂耐药性(P0.01)。细胞外调节蛋白激酶1/2(extracellular regulated protein kinases,ERK1/2)在DU145-mi R-17-92细胞中呈现持续高水平磷酸化,顺铂处理后,其磷酸化水平无明显变化。DU145-mi R-17-92细胞中切除修复互补交叉基因1(excision repair cross complementing1,ERCC1)的m RNA和蛋白质表达水平显著高于DU145-control细胞。结论:高表达mi R-17-92增强了DU145细胞的迁移、侵袭能力,其机制与整合素β1的表达上调及MMP-9活性增强有关。此外,高表达mi R-17-92增强了DU145细胞对顺铂的耐药性,该过程与ERK1/2的磷酸化水平增加和ERCC1的表达水平上调相关。
[Abstract]:Background & AIM: the gene cluster of: mi R-17-92 is closely related to the occurrence of many diseases. It is highly expressed in many tumor cells such as lung cancer, liver cancer, gastric cancer and prostate cancer.The aim of this study was to establish a stable DU145 cell line with high expression of miR-17-92 gene by lentivirus packaging system, and to investigate the effects of mi R-17-92 gene cluster on the migration, invasion and cisplatin resistance of prostate cancer DU145 cells.Methods: the high expression vector of miR-17-92 gene cluster was constructed and transfected into DU145 cell line, and the empty vector was transfected as control. The expression vector was identified by real-time fluorescent quantitative polymerase chain reactionation (RTFQ-PCR).X CELLigence system was used to monitor cell migration, invasion ability and growth after cisplatin treatment, and cell migration was observed by scratch test.Western blotters were used to detect the expression of related proteins and genes by gel zymogram and RTFQ-PCR in order to investigate the mechanism of mi R-17-92 enhancing the migration, invasion and cisplatin resistance of DU145 cells.Results the expression of integrin 尾 1 and the activity of matrix metalloprotein-9 in DU145-control cells were significantly higher than those in DU145-control cells.After treatment with cisplatin, the growth rate of DU145-mi R-17-92 cells was faster than that of DU145-control cells from 12 h.Extracellular regulated protein kinase (1/2(extracellular regulated protein kinasesserine ERK1 / 2) was continuously phosphorylated in DU145-mi R-17-92 cells, and was treated with cisplatin.The expression of m RNA and protein of 1(excision repair cross complementing1ERCC1 in DU145-mi R-17-92 cells was significantly higher than that in DU145-control cells.Conclusion: the overexpression of miR-17-92 enhances the migration and invasion of DU145 cells, and its mechanism is related to the up-regulation of integrin 尾 1 expression and the enhancement of MMP-9 activity.In addition, the overexpression of miR-17-92 enhanced the resistance of DU145 cells to cisplatin, which was related to the increased phosphorylation of ERK1/2 and the up-regulation of ERCC1 expression.
【作者单位】：苏州大学附属第一医院中心实验室;
【基金】：国家自然科学基金资助项目(81172433,81400154) 江苏省自然科学基金资助项目(BK20151211)
【分类号】：R737.25

【相似文献】