miR-448对肺腺癌A549细胞的放疗增敏作用及其机制  

Radiosensitizing effect of miR-448 on lung adenocarcinoma A549 cells and its mechanism

作　　者：余宏伟 张一斌 杨如妃 谢昌斌 廖志伟 周同冲 YU Hongwei;ZHANG Yibin;YANG Rufei;XIE Changbin;LIAO Zhiwei;ZHOU Tongchong(Radiotherapy Department,Cancer Hospital Affiliated to Guangzhou Medical University,Guangzhou 510095,China)

机构地区：[1]广州医科大学附属肿瘤医院放疗科,广州510095

出　　处：《山东医药》2024年第26期5-9,21,共6页Shandong Medical Journal

基　　金：广东省广州市卫生健康科技一般引导项目(20221A010062)。

摘　　要：目的探讨miR-448对肺腺癌A549细胞的放疗增敏作用,并探讨其作用机制。方法取肺腺癌A549细胞,随机分为空白对照组、放疗组、放疗+miR-448-mimics组、放疗+NC-mimics组,放疗+miR-448-mimics组、放疗+NCmimics组先分别转染miR-448-mimics和NC-mimics 24 h,然后给予10 Gy放疗照射,放疗组仅给予放疗照射,空白对照组正常培养。采用MTT法、细胞克隆实验分别检测四组细胞增殖活性和细胞存活情况,Transwell试验检测细胞迁移和侵袭能力,流式细胞术检测细胞凋亡,Western blotting法检测T-框蛋白3(TBX3)、磷酸酶与张力蛋白同源物(PTEN)蛋白,qPCR法检测miR-448及TBX3、PTEN mRNA,双荧光素酶报告试验验证PTEN与miR-448的结合位点。结果与空白对照组比较,放疗组细胞增殖、迁移、侵袭能力降低,细胞凋亡增加(P均<0.05);与放疗组、放疗+NCmimics组比较,放疗+miR-448-mimics组细胞增殖、迁移、侵袭能力降低,细胞凋亡增加(P均<0.05)。与放疗组、放疗+NC-mimics组比较,放疗+miR-448-mimics组细胞PTEN蛋白及mRNA表达升高,TBX3 mRNA表达降低(P均<0.05)。miR-448能够与PTEN的3'UTR特异性结合。结论miR-448可增强肺腺癌A549细胞的放疗敏感性,其机制可能是通过调控TBX3和PTEN表达发挥作用。Objective To investigate the radiosensitizing effect of miR-448 on lung adenocarcinoma A549 cells and to explore its mechanism of action.Methods Lung adenocarcinoma A549 cells were randomly divided into the blank control group,radiotherapy group,radiotherapy+miR-448 mimics group,and radiotherapy+NC mimics group,respectively.Cells in the radiotherapy+miR-448 mimics group and the radiotherapy+NC mimics group were first transfected with miR-448 mimics and NC mimics for 24 h,respectively,and then were irradiated with radiotherapy.Cells in the radiotherapy group were only irradiated with radiotherapy,while cells in the blank control group were cultured normally.MTT assay and cell cloning assay were used to detect the proliferation activity and cell survival of cells in the four groups,Transwell assay was used to detect cell migration and invasion ability,flow cytometry was used to detect the apoptosis,Western blotting was used to detect TBX3 and PTEN proteins,qPCR was used to detect miR-448 and TBX3,phosphatase and tensin homolog(PTEN)mRNA,and dual luciferase assay was used to identify the binding sites between PTEN and miR-448.Results Compared with the blank control group,the cell proliferation,migration and invasion abilities decreased,and the apoptosis increased in the radiotherapy group(all P<0.05).Compared with the radiotherapy group and the radiotherapy+NC mimics group,the cell proliferation,migration and invasion abilities decreased,and the apoptosis increased in the radiotherapy+miR-448 mimics group(all P<0.05).Compared with the radiotherapy group and the radiotherapy+NC mimics group,the expression level of PTEN protein increased in the radiotherapy+miR-448 mimics group(both P<0.05).Compared with the radiotherapy group and the radiotherapy+NC mimics group,the PTEN protein and mRNA levels increased and the TBX3 mRNA level decreased in the radiotherapy+miR-448 mimics group(all P<0.05).MiR-448 specifically bound to the 3'UTR of PTEN.Conclusion The miR-448 can enhance the radiation sensitivity of lung cancer A549

关 键 词：肺肿瘤 肺腺癌细胞 微小核糖核酸 放射治疗 T-框蛋白3 磷酸酶与张力蛋白同源物 

分 类 号：R734.2[医药卫生—肿瘤]