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Beclin 1 通过线粒体依赖途径影响宫颈癌 CaSki 细胞中顺铂诱导的细胞凋亡。

Beclin 1 influences cisplatin-induced apoptosis in cervical cancer CaSki cells by mitochondrial dependent pathway.

机构信息

Department of Obstetrics and Gynecology, Fujian Provincial Hospital, Fujian Provincial Clinical Medical College, Fujian Medical University, Fuzhou, Fujian, China.

出版信息

Int J Gynecol Cancer. 2012 Sep;22(7):1118-24. doi: 10.1097/IGC.0b013e31825e0caa.

Abstract

PURPOSE

To investigate the role of Beclin 1 expression on the cisplatin-induced apoptosis in cervical cancer CaSki cells and to explore the potential mechanism underlying this effect.

MATERIALS AND METHODS

After overexpression or partial silencing of Beclin 1 in cervical cancer CaSki cells, the transfected group and the control group were treated with cisplatin for 24 hours. The percentage of apoptotic cells were assessed by flow cytometry. The mitochondrial membrane potential and activities of caspase-8/9/3 were detected by JC-1 fluorescence staining and colorimetry. The expression of cytochrome c was measured using a Western blot. The messenger RNA expression of Bax and Bcl-2 were detected by real-time quantitative reverse transcription polymerase chain reaction.

RESULTS

Expression of Beclin 1 protein was up-regulated in overexpressed transfectants of CaSki cells. After treatment with cisplatin, the Beclin 1 overexpression group led to the decrease of mitochondrial membrane potential and increase of activities of caspase-9 and caspase-3, and showed a greater increase in apoptosis than did the nontransfected group. Furthermore, Beclin 1 overexpression resulted in increased cytoplasmic cytochrome c and Bax expression and decreased mitochondrial cytochrome c and Bcl-2 expression.

CONCLUSION

Overexpression of Beclin 1 in CaSki cells may influence cisplatin-induced apoptosis by mitochondrial dependent pathway.

摘要

目的

研究 Beclin 1 表达在顺铂诱导宫颈癌 CaSki 细胞凋亡中的作用,并探讨其潜在的作用机制。

材料与方法

在宫颈癌 CaSki 细胞中转染过表达或部分沉默 Beclin 1 后,用顺铂处理转染组和对照组 24 小时。通过流式细胞术评估细胞凋亡的百分比。用 JC-1 荧光染色和比色法检测线粒体膜电位和 caspase-8/9/3 的活性。用 Western blot 法测定细胞色素 c 的表达。用实时定量逆转录聚合酶链反应检测 Bax 和 Bcl-2 的信使 RNA 表达。

结果

Beclin 1 蛋白在 CaSki 细胞过表达转染子中表达上调。用顺铂处理后,Beclin 1 过表达组导致线粒体膜电位降低,caspase-9 和 caspase-3 的活性增加,与未转染组相比,细胞凋亡增加更多。此外,Beclin 1 过表达导致细胞质细胞色素 c 和 Bax 表达增加,线粒体细胞色素 c 和 Bcl-2 表达减少。

结论

Beclin 1 在 CaSki 细胞中的过表达可能通过线粒体依赖性途径影响顺铂诱导的细胞凋亡。

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