本文選題：miR-- + 前列腺腫瘤　； 參考：《中國癌癥雜志》2017年02期

【摘要】：背景與目的:mi R-17-92基因簇與多種疾病的發(fā)生密切相關(guān),其在肺癌、肝癌、胃癌和前列腺癌等多種腫瘤細(xì)胞中均高表達(dá)。本研究利用慢病毒包裝系統(tǒng)建立穩(wěn)定高表達(dá)mi R-17-92基因簇的DU145細(xì)胞株,探討mi R-17-92基因簇對前列腺癌DU145細(xì)胞的遷移、侵襲能力及對順鉑耐藥性的影響。方法:構(gòu)建高表達(dá)mi R-17-92基因簇的表達(dá)載體,轉(zhuǎn)染DU145細(xì)胞株,同時(shí)轉(zhuǎn)染空載體作為對照,并用實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)進(jìn)行鑒定。用x CELLigence系統(tǒng)監(jiān)測細(xì)胞的遷移、侵襲能力及順鉑處理后的生長情況;通過劃痕實(shí)驗(yàn)觀察細(xì)胞的遷移情況;采用蛋白[質(zhì)]印跡法(Western blot)、凝膠酶譜實(shí)驗(yàn)和RTFQ-PCR檢測相關(guān)蛋白質(zhì)和基因的表達(dá)以探討mi R-17-92增強(qiáng)DU145細(xì)胞的遷移、侵襲能力及對順鉑耐藥性的相關(guān)機(jī)制。結(jié)果:DU145-mi R-17-92細(xì)胞遷移速率和侵襲能力高于DU145-control細(xì)胞(P0.01)。DU145-mi R-17-92細(xì)胞中整合素β1的蛋白質(zhì)表達(dá)水平和基質(zhì)金屬蛋白酶-9(matrix metalloprotein-9,MMP-9)的活性顯著高于DU145-control細(xì)胞。順鉑處理后,DU145-mi R-17-92細(xì)胞的生長速度自12 h起快于DU145-control細(xì)胞并呈順鉑耐藥性(P0.01)。細(xì)胞外調(diào)節(jié)蛋白激酶1/2(extracellular regulated protein kinases,ERK1/2)在DU145-mi R-17-92細(xì)胞中呈現(xiàn)持續(xù)高水平磷酸化,順鉑處理后,其磷酸化水平無明顯變化。DU145-mi R-17-92細(xì)胞中切除修復(fù)互補(bǔ)交叉基因1(excision repair cross complementing1,ERCC1)的m RNA和蛋白質(zhì)表達(dá)水平顯著高于DU145-control細(xì)胞。結(jié)論:高表達(dá)mi R-17-92增強(qiáng)了DU145細(xì)胞的遷移、侵襲能力,其機(jī)制與整合素β1的表達(dá)上調(diào)及MMP-9活性增強(qiáng)有關(guān)。此外,高表達(dá)mi R-17-92增強(qiáng)了DU145細(xì)胞對順鉑的耐藥性,該過程與ERK1/2的磷酸化水平增加和ERCC1的表達(dá)水平上調(diào)相關(guān)。
[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.
【作者單位】： 蘇州大學(xué)附屬第一醫(yī)院中心實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(81172433,81400154) 江蘇省自然科學(xué)基金資助項(xiàng)目(BK20151211)
【分類號】：R737.25

【相似文獻(xiàn)】

相關(guān)期刊論文 前1條

1 楊瑞芳;陳麗娟;李建勇;;miR-17-92基因簇與腫瘤[J];中國實(shí)驗(yàn)血液學(xué)雜志;2010年05期

相關(guān)博士學(xué)位論文 前2條

1 李逸塵;MiR-17-92對癌癥網(wǎng)絡(luò)中開關(guān)行為的影響[D];蘭州大學(xué);2011年

2 劉梅;miR-17-92在食管癌中作用機(jī)制的初步研究[D];中國協(xié)和醫(yī)科大學(xué);2009年

相關(guān)碩士學(xué)位論文 前1條

1 馬玉潔;黃牛類胚胎干細(xì)胞分離培養(yǎng)及miR-17-92過表達(dá)對其靶基因表達(dá)的影響[D];廣西大學(xué);2014年

，

本文編號：1765667