【摘要】：目的觀察吉西他濱(gemcitabine,GEM)處理膀胱癌T24細(xì)胞后引起凋亡的同時(shí)是否誘導(dǎo)T24細(xì)胞自噬的產(chǎn)生,并初步探究其自噬產(chǎn)生的機(jī)制,以及自噬與凋亡之間的關(guān)系。方法體外培養(yǎng)T24細(xì)胞,應(yīng)用CCK-8法測(cè)定細(xì)胞抑制率;Western blot檢測(cè)凋亡相關(guān)蛋白Caspase-3、PARP的活化,自噬標(biāo)志蛋白LC3-Ⅱ、底物蛋白P62和自噬潮,以及JNK通路相關(guān)蛋白Jnk1、P-Jnk、Bcl-2的表達(dá)情況;并通過透射電鏡觀察細(xì)胞超微結(jié)構(gòu)自噬體的變化;Annexin V-PI雙流式細(xì)胞術(shù)檢測(cè)抑制自噬后細(xì)胞凋亡率的變化情況。結(jié)果 CCK-8檢測(cè)結(jié)果顯示吉西他濱能有效抑制膀胱癌T24細(xì)胞增殖;1.0μg/mL吉西他濱處理T24細(xì)胞4 h即可發(fā)生明顯的自噬,透射電鏡觀察可見自噬體小體的形成;Western blot檢測(cè)結(jié)果顯示吉西他濱作用T24細(xì)胞后cleaved-Caspase-3、PARP和LC3-Ⅱ表達(dá)增高,P62表達(dá)降低;P-Jnk出現(xiàn)活化,Bcl-2發(fā)生降解;而抑制自噬后能顯著增強(qiáng)吉西他濱誘導(dǎo)的膀胱癌T24細(xì)胞的凋亡。結(jié)論吉西他濱介導(dǎo)膀胱癌T24細(xì)胞凋亡的同時(shí)又誘導(dǎo)保護(hù)性自噬,JNK信號(hào)通路參與調(diào)控其自噬的發(fā)生,抑制自噬可以明顯增強(qiáng)膀胱癌T24細(xì)胞對(duì)吉西他濱的化療敏感性。
[Abstract]:Objective to investigate whether gemcitabine gem can induce apoptosis and induce autophagy of T24 cells, and to explore the mechanism of autophagy and the relationship between autophagy and apoptosis. Methods T24 cells were cultured in vitro. The activation of apoptosis-related protein Caspase-3- 鈪，

本文編號(hào)：2158844