【摘要】：目的:建立Beclin 1過表達和低表達MCF-7細胞模型,檢測4Gy照射后細胞自噬和凋亡的變化,探討B(tài)eclin 1的分子調(diào)控作用機制。方法:實驗分為MCF-7組、MCF-7+4Gy組、MCF-7-Beclin1+4Gy組和MCF-7-Belin1RNAi+4 Gy組。利用分子生物學(xué)方法構(gòu)建Beclin 1過表達載體pcDNA3.1-Beclin 1,并建立Beclin 1過表達和低表達細胞模型。細胞經(jīng)4 Gy照射后,采用MDC染色熒光顯微鏡觀察自噬細胞百分比,AnnexinⅤ-FITC和PI染色流式細胞術(shù)檢測凋亡細胞百分比,Western blotting法檢測Beclin 1、P53、Bcl-2和Bax蛋白表達。結(jié)果:與MCF-7組比較,MCF-7+4Gy組、MCF-7-Beclin 1+4Gy組和MCF-7-Beclin 1RNAi+4Gy組自噬和凋亡細胞百分比均明顯升高(P0.05或P0.01),以MCF-7-Beclin 1+4Gy組升高最明顯,且高于MCF-7+4Gy組(P0.05);各組壞死細胞百分比無明顯差異。4Gy照射后,MCF-7組和MCF-7-Beclin 1組細胞中Beclin 1、P53和Bax蛋白表達水平均升高,Bcl-2蛋白表達水平降低,且以MCF-7-Beclin 1組最為明顯。結(jié)論:成功建立Beclin 1過表達和低表達MCF-7細胞模型,電離輻射能夠誘導(dǎo)其細胞自噬和凋亡,且對Beclin 1過表達細胞作用更明顯。Beclin1通過激活P53,進而抑制Bcl-2和激活Bax,從而形成以P53為中心的自噬與凋亡分子調(diào)控機制。
[Abstract]:Aim: to establish Beclin 1 overexpression and low expression MCF-7 cell model, to detect the changes of autophagy and apoptosis after 4Gy irradiation, and to explore the molecular regulatory mechanism of Beclin 1. Methods: the experiment was divided into MCF-7 group, MCF-7 4Gy group, MCF-7-Beclin1 4Gy group and MCF-7-Belin1RNAi 4 Gy group. Beclin 1 overexpression vector pcDNA3.1-Beclin 1 was constructed by molecular biological method, and Beclin 1 overexpression and low expression cell models were established. After 4 Gy irradiation, the percentage of autophagy cells was observed by MDC staining fluorescence microscope, and the percentage of apoptotic cells was detected by Annexin 鈪，

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