摘要
目的:探讨miR-125b对宫颈癌细胞放射敏感性的影响及其可能的下游机制研究。方法:实时荧光定量聚合酶链反应(quantitative real-time PCR,RT-qPCR)检测宫颈癌组织和细胞中miR-125b和Foxp3的表达量。HeLa细胞进行梯度剂量X射线(0、2、4、6 Gy)照射后,RT-qPCR检测HeLa细胞中miR-125b和Foxp3的表达量。下调miR-125b表达,经6 Gy X射线照射,MTT试验检测HeLa细胞增殖能力,Western blot检测Bax和Bcl-2蛋白表达量。通过Targetscan和双荧光素报告基因实验检测miR-125b与Foxp3的靶向关系。下调Foxp3表达,经6 Gy X射线照射,MTT试验检测HeLa细胞增殖能力,Western blot检测Bax和Bcl-2蛋白表达量。检测miR-125b通过Foxp3对HeLa细胞放射敏感性的影响。下调Foxp3后通过ELISA检测细胞上清液中IL-10和TGF-β的含量。结果:宫颈癌组织和细胞中miR-125b的表达量显著降低,Foxp3表达量显著升高,miR-125b的表达量随着X射线放射剂量增加而升高,Foxp3表达量随着X射线放射剂量增加而降低。6 Gy X射线照射HeLa细胞后,下调miR-125b提高细胞增殖能力,使Bax表达量明显降低,Bcl-2表达量明显升高。miR-125b靶向Foxp3且负调控Foxp3表达。6 Gy X射线照射细胞后,下调Foxp3可显著降低HeLa细胞增殖能力,使Bax表达量明显升高,Bcl-2表达量明显降低。过表达miR-125b能够通过Foxp3增强HeLa细胞的放射敏感性。6 Gy X射线照射细胞后,下调Foxp3降低了细胞中IL-10和TGF-β的表达。结论:上调miR-125b通过靶向和负调控Foxp3,从而增强宫颈癌细胞的放射敏感性,且作用机制可能与下调Foxp3使细胞中IL-10和TGF-β的表达减少有关。
Objective To investigate the effects of miR-125b on radiosensitivity of cervical cancer cells and its possible downstream mechanism.Methods The expression of miR-125b and Foxp3 in cervical cancer tissues and cells was detected by RT-qPCR.The HeLa cells were irradiated with 0,2,4 and 6 Gy of X-rays.The expression of miR-125b and Foxp3 in each group was detected by RT-qPCR.After downregulation of miR-125b expression and 6 Gy X-ray irradiation,the proliferation ability of HeLa cells was detected by MTT assay,and the expression of Bax and Bcl-2 proteins were detected by Western blot.The relationship between miR-125b and Foxp3 was detected by Targetscan and Dual luciferin reporter assay.After downregulation of Foxp3 expression and 6 Gy X-ray irradiation,the proliferation ability of HeLa cells was detected by MTT assay,and the expression of Bax and Bcl-2 proteins were detected by Western blot.The effects of miR-125b on radiosensitivity of HeLa cells through Foxp3 were detected.After down-regulation of Foxp3,the contents of IL-10 and TGF-βin supernatant were detected by ELISA.Results The expression of miR-125b in the tissues and cells of cervical cancer was significantly decreased,while the expression of Foxp3 was significantly increased.The expression of miR-125b in HeLa cells was increased after radiation in a dose dependent manner.The expression of Foxp3 in HeLa cells was decreased after radiation in a dose dependent manner.After 6 Gy X-ray irradiation of HeLa cells,down-regulation of miR-125b increased the cell proliferation capacity,significantly reduced the expression of Bax and increased the expression of Bcl-2.miR-125b targets Foxp3 and negatively regulates Foxp3 expression.After 6 Gy X-ray irradiation of HeLa cells,down-regulation of Foxp3 significantly reduced the proliferation capacity of HeLa cells,increased the expression of Bax and decreased the expression of Bcl-2.Overexpression of miR-125b can enhance radiosensitivity of HeLa cells through Foxp3.After 6 Gy X-ray irradiation,down-regulation of Foxp3 redu
作者
王琳
赵晓华
徐俊
吴衡慧
徐志伟
刘明博
Wang Lin;Zhao Xiaohua;Xu Jun;Wu Henghui;Xu Zhiwei;Liu Mingbo(Department of Gynaecology,Henan Provincial People′s Hospital,Zhengzhou 450003,China;Department of Obstetric center,Qingdao Eighth People′s Hospital,Qingdao 266121,China;Clinical Research Service Center,Henan Provincial People′s Hospital,Zhengzhou 450003,China;Department of Radiotherapy,Henan Provincial People′s Hospital,Zhengzhou 450003,China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2021年第5期361-367,共7页
Chinese Journal of Microbiology and Immunology
