摘要
目的:通过检测Akt联合电离辐射对乳腺癌MCF-7细胞凋亡、自噬和增殖的影响,探讨以Akt为靶点的乳腺癌放射治疗作用。方法:选择人乳腺癌MCF-7细胞、Akt过表达MCF-7(Akt-MCF-7)细胞和Akt低表达(Akt-RNAi-MCF-7)细胞,实验分为MCF-7组、MCF-7+4Gy组、Akt-MCF-7+4Gy组和Akt-RNAi-MCF-7+4Gy组。3种细胞经4Gy照射后,Western blotting法检测Akt蛋白表达,AnnexinⅤ-FITC和PI染色流式细胞术检测细胞凋亡率,MDC染色荧光显微镜观察细胞自噬百分比,MTT法检测细胞增殖活性。结果:与MCF-7细胞比较,Akt-MCF-7细胞中Akt蛋白表达强度增加,Akt-RNAi-MCF-7细胞中Akt蛋白表达强度降低。与MCF-7组比较,MCF-7+4Gy、Akt-MCF-7+4Gy和Akt-RNAi-MCF-7+4Gy组细胞凋亡率和自噬百分比均明显增加(P<0.05或P<0.01),且以Akt-RNAi-MCF-7+4 Gy组增加最明显;与MCF-7+4 Gy组比较,Akt-MCF-7+4Gy组细胞凋亡率和自噬百分比明显降低(P<0.05或P<0.01),而Akt-RNAi-MCF-7+4Gy组细胞凋亡率和自噬百分比明显增加(P<0.05或P<0.01)。与MCF-7组比较,MCF-7+4Gy组、Akt-MCF-7+4Gy组和Akt-RNAi-MCF-7+4 Gy组细胞增殖活性在不同时间点均明显降低(P<0.05或P<0.01),Akt-RNAi-MCF-7+4Gy组细胞增殖活性降至最低。与MCF-7+4Gy组比较,Akt-MCF-7+4Gy组细胞增殖活性在24h时明显升高(P<0.05),而Akt-RNAi-MCF-7+4Gy组细胞增殖活性在不同时间点均明显降低(P<0.01)。结论:Akt过表达可以显著降低电离辐射诱导的MCF-7细胞凋亡、自噬和增殖,而Akt低表达作用则相反。
Objective To measure the effects of Akt combined with ionizing radiation on the apoptosis,autophagy and proliferation in the breast cancer MCF-7cells,and to explore the role of radiotherapy of breast cancer using Akt as a target.Methods The Akt protein expressions in MCF-7,Akt-MCF-7（Akt over-expression）and Akt-RNAiMCF-7（Akt low-expression）cells were detected by Western blotting method after irradiated by 4Gy X-rays,the apoptotic rates were measured by FCM with AnnexinⅤ-FITC and PI staining,the autopahgic cell percentages were measured by fluorescence microscope with MDC staining,and the cell proliferation activities were detected by MTT assay.Results Compared with MCF-7cells,the Akt protein expression in Akt-MCF-7cells was increased,but it was decreased in Akt-RNAi-MCF-7 cells.Compared with MCF-7 group,the apoptotic rates and autophagic percentages in MCF-7＋4Gy,Akt-MCF-7＋4Gy and Akt-RNAi-MCF-7＋4Gy groups were increased significantly（P〈0.05 or P〈0.01）,especially in Akt-RNAi-MCF-7＋4Gy group;compared with MCF-7＋4Gy group,the apoptotic rate and autophagic percentage in Akt-MCF-7＋4Gy group were decreased significantly（P〈0.05 or P〈0.01）,but they were increased significantly in Akt-RNAi-MCF-7＋4Gy group（P〈0.05 or P〈0.01）.Compared with MCF-7group,the cell proliferation activities in MCF-7＋4 Gy group,Akt-MCF-7＋4 Gy group and AktRNAi-MCF-7＋4Gy group were significantly decreased at different time points（P〈0.05 or P〈0.01）,especially in Akt-RNAi-MCF-7＋4Gy group.Compared decreased with MCF-7＋4Gy group,the cell proliferation activity in Akt-MCF-7＋4Gy group was significantly increased at 24h（P〈0.05）,while the cell proliferation activities in Akt-RNAi-MCF-7＋4Gy group were significantly decreased at different time points（P〈0.01）.Conclusion Akt over-expression can significantly reduce the apoptosis,autophagy and proliferation in MCF-7cells induced by ionizing radiation,but Akt low-expression has opposite role.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2015年第1期1-5,I0001,共6页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(30970681)
吉林省教育厅"十二五"科学技术研究项目资助课题(2014-192)
吉林大学白求恩青年科研基金资助课题(2013202017)
关键词
自噬
细胞凋亡
电离辐射
乳腺肿瘤
细胞增殖
autophagy
aopotosis
ionizing radiation
breast neoplasms
cell proliferation
