本文選題：膀胱腫瘤　切入點：microRNA-　出處：《實用醫(yī)學(xué)雜志》2017年20期 　論文類型：期刊論文

【摘要】：目的探討microRNA-497(miR-497)對人膀胱癌EJ細(xì)胞增殖和凋亡的影響。方法將人膀胱癌細(xì)胞株EJ分為4組:miR-497 mimics、mimics NC、miR-497 inhibitor、inhibitor NC;通過脂質(zhì)體Lipfectami-neTM2000介導(dǎo)將miR-497 mimics、mimics NC、miR-497 inhibitor、inhibitor NC瞬時轉(zhuǎn)染至上述4組細(xì)胞中,轉(zhuǎn)染6 h后,在熒光顯微鏡下觀察轉(zhuǎn)染效率;轉(zhuǎn)染48h后,采用qRT-PCR技術(shù)檢測各組細(xì)胞中miR-497的表達(dá);平板克隆實驗和噻唑藍(lán)(MTT)實驗檢測細(xì)胞增殖情況,流式細(xì)胞術(shù)檢測細(xì)胞凋亡情況,Western blot法檢測凋亡相關(guān)蛋白Bcl-2和Caspase-3的表達(dá)變化。結(jié)果熒光顯微鏡下觀察結(jié)果顯示:各組細(xì)胞轉(zhuǎn)染效率均90%;qRT-PCR結(jié)果顯示:與各自陰性對照組相比,轉(zhuǎn)染miR-497 mimics組,EJ細(xì)胞中miR-497表達(dá)明顯增加(P0.01),而轉(zhuǎn)染miR-497 inhibitor后表達(dá)明顯下調(diào)(P0.01)。過表達(dá)miR-497后,EJ細(xì)胞的增殖能力較陰性對照組明顯減弱(P0.01),凋亡率明顯上升(P0.01);而抑制miR-497表達(dá)后,EJ細(xì)胞的增殖能力較陰性對照組明顯增強(qiáng)(P0.05),凋亡率明顯降低(P0.01)。Western blot結(jié)果示:與陰性對照組相比,過表達(dá)miR-497可使EJ細(xì)胞的Bcl-2表達(dá)降低(P0.01),Caspase-3表達(dá)增加(P0.01)。結(jié)論miR-497可抑制膀胱癌EJ細(xì)胞增殖,促進(jìn)其凋亡,可能與影響B(tài)cl-2和Caspase-3蛋白表達(dá)有關(guān)。
[Abstract]:Objective to investigate the effect of microRNA-497miR-497) on the proliferation and apoptosis of human bladder cancer EJ cells. Methods Human bladder cancer cell line EJ was divided into 4 groups: 1: miR-497 mimics nCmmiR-497 inhibitor inhibitor NC.The miR-497 mimicsmimics NCmiR-497 inhibitory inhibitor NC was transiently transfected into the above 4 cells by liposome Lipfectami-neTM2000 for 6 h after transfection. The transfection efficiency was observed under fluorescence microscope, the expression of miR-497 in each group was detected by qRT-PCR technique after 48 h transfection, the proliferation of cells was detected by plate cloning assay and thiazolyl blue qRT-PCR assay. The expression of apoptosis-related protein Bcl-2 and Caspase-3 was detected by flow cytometry and Western blot. Results the transfection efficiency of each group was 90g / qRT-PCR, which was higher than that of the control group (P < 0.05). After transfection of miR-497 mimics, the expression of miR-497 in EJ cells increased significantly, while that of miR-497 inhibitor decreased significantly after transfection of miR-497 inhibitor. The proliferative ability of EJ cells after overexpression of miR-497 was significantly lower than that of negative control group, and the apoptotic rate of EJ cells increased significantly, while that of EJ cells after inhibiting miR-497 expression was significantly higher than that of negative control group. Compared with the negative control group, the proliferation ability of the cells was significantly increased, and the apoptosis rate was significantly decreased. The results of Western blot showed that: compared with the negative control group, the cell apoptosis rate was significantly lower than that in the negative control group. Overexpression of miR-497 could decrease the expression of Bcl-2 in EJ cells and increase the expression of Caspase-3. Conclusion miR-497 can inhibit the proliferation and promote the apoptosis of EJ cells, which may be related to the expression of Bcl-2 and Caspase-3.
【作者單位】： 南方醫(yī)科大學(xué)研究生學(xué)院;南方醫(yī)科大學(xué)附屬廣州軍區(qū)廣州總醫(yī)院泌尿外科;南方醫(yī)科大學(xué)附屬廣州軍區(qū)廣州總醫(yī)院婦產(chǎn)科;
【基金】：國家自然科學(xué)基金資助項目(編號:81372744) 廣東省自然科學(xué)基金資助項目(編號:2016A030313616) 廣州市科技基金資助項目(編號:201607010184)
【分類號】：R737.14

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