中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2011年
10期
742-746
,共5页
兰海%林丛尧%袁宏银%熊斌
蘭海%林叢堯%袁宏銀%熊斌
란해%림총요%원굉은%웅빈
癌,非小细胞肺%miR-21%程序性细胞死亡因子4%细胞增殖%细胞凋亡
癌,非小細胞肺%miR-21%程序性細胞死亡因子4%細胞增殖%細胞凋亡
암,비소세포폐%miR-21%정서성세포사망인자4%세포증식%세포조망
Carcinoma,non-small cell lung%miR-21%Programmed cell death 4%Cell proliferation%Apoptosis
目的 研究非小细胞肺癌(NSCLC)中高表达的miR-21对细胞增殖和凋亡的影响及其调控机制.方法 采用荧光定量聚合酶链反应检测miR-21在人NSCLC组织、癌旁组织和A549细胞系中的表达.通过序列分析预测被miR-21调控的抑癌基因,通过荧光素酶活性检测和Western blot 检测验证miR-21对靶基因表达调控的影响.采用RNA干扰技术抑制miR-21和程序性细胞死亡因子4(PDCD4),以台盼蓝染色和流式细胞术检测A549细胞增殖和凋亡的变化.结果 NSCLC组织和A549细胞中miR-21的表达水平分别为癌旁组织的2.24倍和3.06倍.序列预测和基因表达调控研究表明,miR-21可以在NSCLC中反向调控PDCD4的表达(P＜0.01).荧光素酶活性检测结果显示,共同转入Wt 3'UTR和miR-21会显著抑制A549细胞中的荧光素酶表达(P＜O.001).Western blot检测结果显示,导入反义寡核苷酸抑制miR-21的功能后,PDCD4的表达水平明显上升.抑制miR-21的作用可以显著抑制A549细胞的增殖,并使细胞凋亡率由2.6％上升到10.9％,而抑制PDCD4的表达可以在很大程度上消除miR-21介导的细胞增殖障碍,抑制细胞凋亡.结论 在NSCLC中,抑制抑癌基因PDCD4的表达可能是miR-21介导肿瘤细胞增殖、抵抗凋亡的重要途径之一.
目的 研究非小細胞肺癌(NSCLC)中高錶達的miR-21對細胞增殖和凋亡的影響及其調控機製.方法 採用熒光定量聚閤酶鏈反應檢測miR-21在人NSCLC組織、癌徬組織和A549細胞繫中的錶達.通過序列分析預測被miR-21調控的抑癌基因,通過熒光素酶活性檢測和Western blot 檢測驗證miR-21對靶基因錶達調控的影響.採用RNA榦擾技術抑製miR-21和程序性細胞死亡因子4(PDCD4),以檯盼藍染色和流式細胞術檢測A549細胞增殖和凋亡的變化.結果 NSCLC組織和A549細胞中miR-21的錶達水平分彆為癌徬組織的2.24倍和3.06倍.序列預測和基因錶達調控研究錶明,miR-21可以在NSCLC中反嚮調控PDCD4的錶達(P＜0.01).熒光素酶活性檢測結果顯示,共同轉入Wt 3'UTR和miR-21會顯著抑製A549細胞中的熒光素酶錶達(P＜O.001).Western blot檢測結果顯示,導入反義寡覈苷痠抑製miR-21的功能後,PDCD4的錶達水平明顯上升.抑製miR-21的作用可以顯著抑製A549細胞的增殖,併使細胞凋亡率由2.6％上升到10.9％,而抑製PDCD4的錶達可以在很大程度上消除miR-21介導的細胞增殖障礙,抑製細胞凋亡.結論 在NSCLC中,抑製抑癌基因PDCD4的錶達可能是miR-21介導腫瘤細胞增殖、牴抗凋亡的重要途徑之一.
목적 연구비소세포폐암(NSCLC)중고표체적miR-21대세포증식화조망적영향급기조공궤제.방법 채용형광정량취합매련반응검측miR-21재인NSCLC조직、암방조직화A549세포계중적표체.통과서렬분석예측피miR-21조공적억암기인,통과형광소매활성검측화Western blot 검측험증miR-21대파기인표체조공적영향.채용RNA간우기술억제miR-21화정서성세포사망인자4(PDCD4),이태반람염색화류식세포술검측A549세포증식화조망적변화.결과 NSCLC조직화A549세포중miR-21적표체수평분별위암방조직적2.24배화3.06배.서렬예측화기인표체조공연구표명,miR-21가이재NSCLC중반향조공PDCD4적표체(P＜0.01).형광소매활성검측결과현시,공동전입Wt 3'UTR화miR-21회현저억제A549세포중적형광소매표체(P＜O.001).Western blot검측결과현시,도입반의과핵감산억제miR-21적공능후,PDCD4적표체수평명현상승.억제miR-21적작용가이현저억제A549세포적증식,병사세포조망솔유2.6％상승도10.9％,이억제PDCD4적표체가이재흔대정도상소제miR-21개도적세포증식장애,억제세포조망.결론 재NSCLC중,억제억암기인PDCD4적표체가능시miR-21개도종류세포증식、저항조망적중요도경지일.
Objective To elucidate the regulatory mechanism underlying proliferation and antiapoptosis in NSCLC by overexpression of miR-21.Methods Real-time PCR was used to measure miR-21 abundance in non-small cell lung cancer(NSCLC)tumor samples and adjacent normal tissues,as well as NSCLC cell lines.Tumor suppressor genes as potential targets of miR-21 were predicted by sequence analysis.Luciferase assay and Western blot were used to assess the regulatory effect.The effect on A549 cell viability and apoptosis by miR-21-induced gene repression was tested by trypan-blue exclusion and flow cytometry.Results miR-21 expression was 2.24-fold higher in the NSCLC tumor samples and 3.06-fold higher in the A549 cells than that in the adjacent normal tissues.Sequence prediction and gene expression regulation assays showed that miR-21 could reversely regulate the expression of PDCD4(P ＜ 0.01).Suppression of miR-21 expression is associated with an elevation of Pdcd4,resulting in a significant reduction of proliferation and the apoptosis rate(2.6％)was increased to 10.9％.Moreover,the antiproliferation and pro-apoptotic effect by miR-21 suppression could be reversed by PDCD4 knock down.Conclusion Suppression of the tumor suppressor PDCD4 expression may be one of the important regulatory pathways of the miR-21-mediated cell proliferation and decrease of apoptosis in non-small cell lung cancer.
