本文選題：PLCε　切入點(diǎn)：miR-　出處：《中國(guó)生物工程雜志》2017年03期 　論文類型：期刊論文

【摘要】：目的:探討miR-145調(diào)控PLCε對(duì)膀胱癌細(xì)胞T24上皮間質(zhì)轉(zhuǎn)化(EMT)和遷移的影響及可能的分子機(jī)制。方法:(1)腺病毒感染T24細(xì)胞,劃痕實(shí)驗(yàn)和Transwell檢測(cè)細(xì)胞的遷移能力;RTPCR、Western blot分別檢測(cè)PLCε及EMT相關(guān)分子的表達(dá);為探究其分子機(jī)制,Western blot檢測(cè)GSK-3β磷酸化(Ser9位點(diǎn))和Snail的表達(dá)情況。(2)利用生物信息學(xué)技術(shù)預(yù)測(cè)可能調(diào)控PLCε的miRNA,結(jié)合文獻(xiàn)報(bào)道的膀胱癌microRNA表達(dá)譜結(jié)果篩選出miR-145;轉(zhuǎn)染miR-145 mimics至T24,q PCR檢測(cè)miR-145、PLCε的表達(dá),Western blot檢測(cè)PLCε的表達(dá)。(3)轉(zhuǎn)染miR-145 mimics,Western blot檢測(cè)EMT相關(guān)分子及p-GSK-3β、Snail;劃痕實(shí)驗(yàn)、Transwell檢測(cè)過(guò)表達(dá)miR-145后細(xì)胞的遷移能力。結(jié)果:(1)干擾PLCε表達(dá)能顯著抑制細(xì)胞的遷移,同時(shí),使T24細(xì)胞中E-cadherin表達(dá)上調(diào),N-cadherin和Vimentin表達(dá)下調(diào);干擾PLCε后,GSK-3β磷酸化(Ser9位點(diǎn))水平下降,Snail表達(dá)降低。(2)轉(zhuǎn)染miR-145 mimics可使T24細(xì)胞中miR-145表達(dá)增高,且明顯抑制T24細(xì)胞中PLCε的表達(dá)。(3)在T24中過(guò)表達(dá)miR-145,細(xì)胞遷移能力顯著下降,EMT標(biāo)志分子的表達(dá)情況與沉默PLCε結(jié)果一致。同時(shí),與陰性對(duì)照組相比,轉(zhuǎn)染miR-145 mimics組p-GSK-3β和Snail表達(dá)顯著減少。結(jié)論:PLCε通過(guò)GSK-3β/Snail信號(hào)通路促進(jìn)膀胱癌細(xì)胞T24發(fā)生EMT及遷移,miR-145可以逆轉(zhuǎn)PLCε誘導(dǎo)膀胱癌EMT的發(fā)生,從而阻止膀胱癌細(xì)胞的遷移。
[Abstract]:Objective: to investigate the effect and possible molecular mechanism of miR-145 regulating PLC 蔚 on the epithelial mesenchymal transformation and migration of bladder cancer cell line T24. Methods: T24 cells were infected with adenovirus. The migration ability of cells was detected by scratch test and Transwell. The expression of PLC 蔚 and EMT related molecules were detected by RT PCR Western blot. In order to explore its molecular mechanism, the expression of GSK-3 尾 phosphorylation site Ser9) and Snail were detected by Western blot) and the miRNAs that might regulate PLC 蔚 were predicted by bioinformatics. MiR-145 was screened according to the reported results of microRNA expression in bladder cancer and transfected with miR-145 mimics. T24Q PCR was used to detect the expression of miR-145P PLC 蔚. Western blot was used to detect the expression of PLC 蔚.) transfection miR-145 mimicsl Western blot was used to detect EMT related molecules and p-GSK-3 尾 Snail.The scratch test was used to detect the migration ability of the cells after miR-145 expression. At the same time, the expression of E-cadherin in T24 cells was up-regulated and the expression of N-cadherin and Vimentin was down-regulated, and the level of GSK-3 尾 phosphorylated ser9 was decreased after interfering with PLC 蔚. The expression of miR-145 in T24 cells was increased by transfection of miR-145 mimics. The overexpression of miR-145in T24 cells was significantly inhibited, and the expression of PLC markers was significantly decreased in T24 cells, which was consistent with the results of silencing PLC 蔚. At the same time, compared with the negative control group, the expression of miR-145in T24 cells was similar to that in the negative control group. Conclusion the expression of p-GSK-3 尾 and Snail decreased significantly in miR-145 mimics transfection group. Conclusion the expression of p-GSK-3 尾 and Snail in T24 cells induced by GSK-3 尾 / Snail signal pathway can be reversed by PLC 蔚 -induced EMT, thus preventing the migration of T24 cells.
【作者單位】： 重慶醫(yī)科大學(xué)檢驗(yàn)醫(yī)學(xué)院臨床檢驗(yàn)診斷學(xué)教育部重點(diǎn)實(shí)驗(yàn)室重慶市重點(diǎn)實(shí)驗(yàn)室;重慶醫(yī)科大學(xué)附屬第一醫(yī)院泌尿外科;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(81072086)
【分類號(hào)】：R737.14

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