摘要
目的探讨miR-6732-3p表达对人鼻咽癌CNE-2R细胞放射敏感性的影响。方法采用RT-qPCR检测miR-6732-3p在CNE-2和CNE-2R细胞株中的表达。用miR-6732-3p慢病毒转染CNE-2R细胞后,将CNE-2R细胞分为三组,分别为CNE-2R组(正常对照组)、miR-6732-3p NC组(转染对照组)、miR-6732-3p inhibition组(转染组)。RT-qPCR检测各组细胞中miR-6732-3p的表达,CCK-8、流式细胞术和克隆形成实验分别检测转染前后的细胞增殖、凋亡能力及放射敏感性。结果RT-qPCR检测结果显示,miR-6732-3p在CNE-2R细胞中的表达量明显高于CNE-2细胞(P=0.036)。采用慢病毒转染CNE-2R细胞后,转染组细胞miR-6732-3p的表达量均较转染对照组和正常对照组低(P<0.05)。CCK-8法检测结果显示,与两个对照组相比,转染组细胞生长受抑制(P<0.001);在不同照射剂量下,转染组细胞的存活分数均明显降低(P<0.001)。流式细胞仪检测结果显示,未经照射(0 Gy)和接受8 Gy照射后转染组细胞凋亡率均较转染对照组及正常对照组明显升高(P<0.05)。克隆形成实验结果显示,转染组的放射生物学参数D0、Dq、SF2值均低于两个对照组(P<0.05);在不同照射剂量下,转染组细胞存活分数亦低于两个对照组(P<0.001)。结论沉默miR-6732-3p可提高鼻咽癌CNE-2R细胞的放射敏感性,为鼻咽癌放射抗拒的分子靶向治疗提供新思路。
Objective To investigate the effect of miR-6732-3p expression on the radio-sensitivity of human nasopharyngeal carcinoma CNE-2R cells.Methods The expression of miR-6732-3p in CNE-2 and CNE-2R cell lines was detected by RT-qPCR.After transfection of CNE-2R cells with miR-6732-3p lentivirus,CNE-2R cells were divided into CNE-2R(normal control group),miR-6732-3p NC(transfection control group),and miR-6732-3p inhibition(transfection group).The expression of miR-6732-3p in each group was detected by RT-qPCR.The proliferation ability,apoptosis ability and radio-sensitivity of cells before and after transfection were detected by CCK-8 assay,flow cytometry and cloning formation assay,respectively.Results The RT-q PCR showed that the expression level of miR-6732-3p in CNE-2R cells was significantly higher than that of CNE-2 cells(P=0.036).After transfection of CNE-2R cells with lentivirus,the expression level of miR-6732-3p in the transfection group was lower than those in the transfection control group and the normal control group(P<0.05).CCK-8 assay results showed that compared with the two control groups,the cell growth of the transfection group was inhibited(P<0.001);the survival fraction(SF)of the transfection group was significantly reduced under different irradiation doses(P<0.001).The flow cytometry detection results showed that the apoptotic rate of transfection group after non-irradiation(0 Gy)and 8 Gy irradiation was significantly increased than that of the transfection control group and the normal control group(P<0.05).The cloning formation assay indicated that the radiobiological parameters D0,Dq and SF2 in the transfection group were lower than those of the two control groups(P<0.05);the SF of the transfection group was also lower than the two control groups under different irradiation doses(P<0.001).Conclusions Silencing miR-6732-3p expression can significantly improve the radio-sensitivity of nasopharyngeal carcinoma CNE-2R cells,providing a new idea for molecular targeted therapy of radio-resistance in na
作者
杨晓慧
李开国
朱小东
李龄
杜有芹
杨柳
曲颂
YANG Xiaohui;LI Kaiguo;ZHU Xiaodong;LI Ling;DU Youqin;YANG Liu;QU Song(Department of Radiotherapy,Guangxi Medical University Cancer Hospital,Guangxi Nasopharyngeal Carcinoma Clinical Medical Research Center,Nanning 530021,China)
出处
《中国癌症防治杂志》
CAS
2020年第4期403-408,共6页
CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
基金
广西自然科学基金项目(2018GXNSFAA281032)
广西医疗卫生适宜技术开发与推广应用项目(S2018038)
广西研究生教育创新计划项目(YCSW2019120)
区域性高发肿瘤早期防治研究教育部重点实验室(广西医科大学)自主研究项目(GK2018-08
GKE2019-15)。
