發(fā)布時(shí)間：2018-05-09 15:10

  本文選題：膀胱癌 + MEXA基因��； 參考：《解剖學(xué)報(bào)》2017年06期

【摘要】：目的探討通過慢病毒介導(dǎo)的RNA干擾技術(shù)抑制MEX3A基因表達(dá)對(duì)膀胱癌細(xì)胞增殖和凋亡的影響。方法應(yīng)用GV115載體構(gòu)建針對(duì)MEX3A基因的shRNA慢病毒載體,轉(zhuǎn)染包裝細(xì)胞293T產(chǎn)生慢病毒顆粒,收集并滴度測(cè)定后轉(zhuǎn)染5637膀胱癌細(xì)胞,實(shí)驗(yàn)組轉(zhuǎn)染MEX3A-shRNA慢病毒,對(duì)照組轉(zhuǎn)染陰性對(duì)照慢病毒;實(shí)時(shí)定量PCR(Real-time PCR)檢測(cè)MEX3A基因敲減效率;慢病毒轉(zhuǎn)染3 d后,使用Celigo連續(xù)細(xì)胞計(jì)數(shù)5d檢測(cè)細(xì)胞生長(zhǎng);慢病毒轉(zhuǎn)染5d后,進(jìn)行膜聯(lián)蛋白V(Annexin V)染色并用流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡。結(jié)果成功構(gòu)建MEX3AshRNA慢病毒載體以及穩(wěn)定抑制MEX3A表達(dá)的細(xì)胞株,MEX3A基因敲減效率達(dá)到74%;Celigo細(xì)胞計(jì)數(shù)檢測(cè)顯示,與對(duì)照組相比,實(shí)驗(yàn)組5637細(xì)胞的增殖速率受到顯著抑制;流式細(xì)胞術(shù)檢測(cè)顯示,實(shí)驗(yàn)組發(fā)生凋亡的5637細(xì)胞顯著增加。結(jié)論 MEX3A基因促進(jìn)膀胱癌細(xì)胞的增殖,抑制膀胱癌細(xì)胞的凋亡。
[Abstract]:Objective to investigate the effect of lentivirus-mediated RNA interference on the proliferation and apoptosis of bladder cancer cells. Methods the shRNA lentivirus vector targeting MEX3A gene was constructed with GV115 vector. Lentivirus particles were produced by transfection of packaging cells 293T. After collecting and titer determination, 5637 bladder cancer cells were transfected with lentivirus. The experimental group transfected MEX3A-shRNA lentivirus and the control group transfected negative control lentivirus. MEX3A gene knockout efficiency was detected by real-time quantitative PCR(Real-time PCR, cell growth was detected by Celigo continuous cell count for 5 days after lentivirus transfection for 3 days, and cell apoptosis was detected by flow cytometry with V(Annexin V staining after 5 days of lentivirus transfection. Results the successful construction of MEX3AshRNA lentivirus vector and the knockdown efficiency of MEX3A gene reached 74%. Compared with the control group, the proliferation rate of 5637 cells in the experimental group was significantly inhibited. Flow cytometry showed that the number of apoptotic 5637 cells in the experimental group was significantly increased. Conclusion MEX3A gene can promote the proliferation of bladder cancer cells and inhibit the apoptosis of bladder cancer cells.
【作者單位】： 中國(guó)醫(yī)科大學(xué)附屬盛京醫(yī)院超聲科;
【基金】：國(guó)家自然科學(xué)基金(81371552)
【分類號(hào)】：R737.14

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