Akt联合电离辐射对人乳腺癌MCF-7细胞凋亡、自噬和增殖的影响

doi:10.13481/j.1671-587x.20150101

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (01): 1-5.doi: 10.13481/j.1671-587x.20150101

• 基础研究 • 下一篇

Akt联合电离辐射对人乳腺癌MCF-7细胞凋亡、自噬和增殖的影响

齐亚莉^1,2, 赵大力³, 刘彦军⁴, 龚守良², 王志成²

1. 北华大学公共卫生学院流行病学教研室, 吉林吉林 132011;
2. 吉林大学公共卫生学院卫生部放射生物学重点实验室, 吉林长春 130021;
3. 吉林(市)出入境检验检疫局, 吉林吉林 132011;
4. 吉林省长春市人民医院放疗科, 吉林长春 130061

收稿日期:2014-08-27 出版日期:2015-01-28 发布日期:2015-01-30
通讯作者: 王志成,讲师(Tel:0431-85619443,E-mail:zhicheng@jlu.edu.cn) E-mail:zhicheng@jlu.edu.cn
作者简介:齐亚莉(1973-),女,吉林省四平市人,副教授,医学博士,主要从事艾滋病和肿瘤流行病学方面的研究。
基金资助:
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)

Effects of Akt combined with ionizing radiation onapoptosis, autophagy and proliferation in breast cancer MCF-7 cells

QI Yali^1,2, ZHAO Dali³, LIU Yanjun⁴, GONG Shouliang², WANG Zhicheng²

1. Department of Epidemiology, School of Public Health, Beihua University, Jilin 132001, China;
2. Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China;
3. Jilin Province (City) Entry and Exit Inspection and Quarantine Bureau, Jilin 132011, China;
4. Department of Radiotherapy, People's Hospital of Changchun City, Changchun 130061, China

Received:2014-08-27 Online:2015-01-28 Published:2015-01-30

摘要/Abstract

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+4 Gy组、Akt-MCF-7+4 Gy组和Akt-RNAi-MCF-7+ 4 Gy组。3种细胞经4 Gy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+4 Gy、Akt-MCF-7+4 Gy和Akt-RNAi-MCF-7+4 Gy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+4 Gy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+4 Gy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+4 Gy组、Akt-MCF-7+4 Gy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+4 Gy组细胞增殖活性降至最低。与MCF-7+4 Gy组比较,Akt-MCF-7+4 Gy组细胞增殖活性在24 h时明显升高(P<0.05),而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<0.01)。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。

关键词: 自噬, 细胞凋亡, 电离辐射, 乳腺肿瘤, 细胞增殖

Abstract:

Objective To measure the effects of Akt combined with ionizing radiation on the apoptosis, autophagy and proliferation in the breast cancer MCF-7 cells, 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-RNAi-MCF-7 (Akt low-expression) cells were detected by Western blotting method after irradiated by 4 Gy 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-7 cells, the Akt protein expression in Akt-MCF-7 cells 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+4 Gy, Akt-MCF-7+4 Gy and Akt-RNAi-MCF-7+4 Gy groups were increased significantly (P<0.05 or P<0.01), especially in Akt-RNAi-MCF-7+4 Gy group; compared with MCF-7+4 Gy group, the apoptotic rate and autophagic percentage in Akt-MCF-7+4 Gy group were decreased significantly (P<0.05 or P<0.01), but they were increased significantly in Akt-RNAi-MCF-7+4 Gy group (P<0.05 or P<0.01).Compared with MCF-7 group, the cell proliferation activities in MCF-7+4 Gy group, Akt-MCF-7+4 Gy group and Akt-RNAi-MCF-7+4 Gy group were significantly decreased at different time points(P<0.05 or P<0.01), especially in Akt-RNAi-MCF-7+4 Gy group.Compared decreased with MCF-7+4 Gy group, the cell proliferation activity in Akt-MCF-7+4 Gy group was significantly increased at 24 h (P<0.05), while the cell proliferation activities in Akt-RNAi-MCF-7+4 Gy 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-7 cells induced by ionizing radiation, but Akt low-expression has opposite role.

Key words: autophagy, aopotosis, ionizing radiation, breast neoplasms, cell proliferation

中图分类号:

R730.55

齐亚莉, 赵大力, 刘彦军, 龚守良, 王志成. Akt联合电离辐射对人乳腺癌MCF-7细胞凋亡、自噬和增殖的影响[J]. 吉林大学学报(医学版), 2015, 41(01): 1-5.

QI Yali, ZHAO Dali, LIU Yanjun, GONG Shouliang, WANG Zhicheng. Effects of Akt combined with ionizing radiation onapoptosis, autophagy and proliferation in breast cancer MCF-7 cells[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 1-5.

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Akt联合电离辐射对人乳腺癌MCF-7细胞凋亡、自噬和增殖的影响

Effects of Akt combined with ionizing radiation onapoptosis, autophagy and proliferation in breast cancer MCF-7 cells

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