本文關(guān)鍵詞： miR- 程序性細(xì)胞死亡因子 -O細(xì)胞 增殖 凋亡　出處：《西安交通大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年06期 　論文類型：期刊論文

【摘要】：目的探討miR-21在腎透明細(xì)胞癌中的表達(dá)及其臨床意義,以及如何通過調(diào)節(jié)程序性細(xì)胞死亡因子4(programmed cell death 4,PDCD4)的表達(dá)影響786-O腎透明細(xì)胞癌細(xì)胞系的增殖和凋亡。方法通過分析The Cancer Genome Atlas(TCGA)腎透明細(xì)胞癌數(shù)據(jù)庫,比較癌組織及正常癌旁組織中miR-21的表達(dá)水平;分析miR-21表達(dá)水平在不同臨床病理分期腎癌組織中的差異;采用Kaplan-Meier法和對數(shù)秩和檢驗(Log-rank test)研究miR-21表達(dá)水平和患者生存之間的關(guān)系;通過轉(zhuǎn)染miR-21抑制性核苷酸(AS-miR-21)下調(diào)miR-21表達(dá)水平,采用MTT和流式細(xì)胞術(shù)分別檢測細(xì)胞增殖和凋亡,采用實時定量PCR(qRT-PCR)和Western blot測量PDCD4mRNA和蛋白質(zhì)表達(dá)水平變化,采用雙熒光素報告系統(tǒng)檢測miR-21對PDCD4的直接調(diào)節(jié)。結(jié)果腎透明細(xì)胞癌組織中miR-21的表達(dá)水平顯著高于癌旁組織(P0.000 1)。miR-21在Ⅲ期和Ⅳ期腎癌組織中表達(dá)水平顯著高于Ⅰ期(P均0.000 1),miR-21表達(dá)水平與臨床病理分期呈正相關(guān)(r=0.262,P0.000 1)。miR-21表達(dá)水平與T分期(r=0.250,P0.000 1)與淋巴結(jié)轉(zhuǎn)移陽性(N1)以及遠(yuǎn)處轉(zhuǎn)移均呈正相關(guān)(P均0.001)。生存分析顯示miR-21高表達(dá)患者中位生存時間顯著短于miR-21低表達(dá)者中位生存時間(Log-rank,P0.001)。下調(diào)miR-21后,786-O細(xì)胞的增殖能力較對照顯著降低(P0.05),凋亡顯著增加(P=0.005),PDCD4mRNA(P=0.002)和蛋白質(zhì)表達(dá)水平顯著增高。雙熒光素報告實驗顯示在轉(zhuǎn)染AS-miR-21的細(xì)胞內(nèi)PDCD4相對熒光強度較對照細(xì)胞顯著升高(P=0.003)。結(jié)論 miR-21在腎透明細(xì)胞癌組織中表達(dá)升高,與患者臨床病理分期呈正相關(guān),和患者生存呈負(fù)相關(guān);miR-21可能通過調(diào)節(jié)PDCD4表達(dá)水平,參與調(diào)節(jié)腎透明細(xì)胞癌細(xì)胞的增殖和凋亡。
[Abstract]:Objective to investigate the expression of miR-21 in renal clear cell carcinoma and its clinical significance. And how to influence the proliferation and apoptosis of 786-O renal clear cell carcinoma cell line by regulating the expression of programmed cell death factor 4programed cell death 4 PDCD4.Methods by analyzing the The Cancer Genome Atlas TCGA-based renal clear cell carcinoma database, To compare the expression of miR-21 in cancer tissues and normal tissues, to analyze the difference of miR-21 expression in renal cell carcinoma with different clinicopathologic stages, to study the relationship between the expression of miR-21 and the survival of patients by Kaplan-Meier and log-rank test. The expression of miR-21 was down-regulated by transfection of miR-21 inhibitory nucleotide AS-miR-21. Cell proliferation and apoptosis were detected by MTT and flow cytometry, and the expression of PDCD4mRNA and protein were measured by real-time quantitative PCRQ RT-PCR and Western blot. Double fluorescein reporting system was used to detect the direct regulation of PDCD4 by miR-21. Results the expression level of miR-21 in renal clear cell carcinoma tissues was significantly higher than that in paracancerous tissues (P 0.000 1) .miR-21 in stage 鈪，

本文編號：1539082