miR-1269a表达对H460干细胞样细胞放疗敏感性的影响  

Effect of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell like cells

作　　者：姜贻乾[1] 郭庆敏[1] 朱春兰 徐小平[1] 安素红[1] 鲍艳红 苏芳[1] 俞灿炳 Jiang Yi-Qian;Guo Qing-Min;Zhu Chun-Lan;Xu Xiao-Ping;An Su-Hong;Bao Yan-Hong;Su Fang;Yu Can-Bing(Department of Radiotherapy,the First People's Hospital of Xiaoshan District,Hangzhou 311200,China)

机构地区：[1]杭州市萧山区第一人民医院放疗科,杭州311200

出　　处：《解放军医学杂志》2020年第9期922-928,共7页Medical Journal of Chinese People's Liberation Army

基　　金：杭州市科技计划项目(20171226Y82);杭州市卫生计划项目(2017B16)。

摘　　要：目的研究miR-1269a表达对非小细胞肺癌H460干细胞样细胞放疗敏感性的影响。方法选取非小细胞肺癌H460细胞系进行无血清悬浮培养,通过流式细胞仪筛选CD133及CD44阳性的H460干细胞样细胞。通过RT-qPCR检测miR-30a、miR-148a、miR-148b、miR-152、miR-411、miR-497、miR-598、miR-424、miR-761、miR-1269a、miR-1280及CD44、CD133 mRNA在H460干细胞样细胞内的表达水平。采用CCK-8实验、干细胞成球实验、流式细胞术及彗星实验分析miR-1269a表达对H460干细胞样细胞放疗敏感性的影响。结果H460干细胞样细胞的CD133及CD44 mRNA表达水平较H460细胞明显增加,分别为H460细胞的(1.30±0.03)倍和(1.19±0.02)倍(P<0.05)。miR-1269a在H460干细胞样细胞中的表达量最高,且高于H460细胞中的表达量,差异有统计学意义(P<0.05)。转染miR-1269a inhibitor后经X线照射处理,H460干细胞样细胞的生长抑制率(72.11%±2.45%)明显增高,其凋亡率达17.70%±0.54%,细胞内DNA双链断裂也明显增加,在2 Gy的X线辐射处理后DNA尾矩为转染前的(1.19±0.02)倍,即转染miR-1269a inhibitor明显增加了H460干细胞样细胞对放疗的敏感性(P<0.05)。转染pcDNA3.1-SOX7后经X线照射处理对H460干细胞样细胞的作用与转染miR-1269a inhibitor相近。转染miR-1269a mimics可逆转pcDNA3.1-SOX7对H460干细胞样细胞恶性生物学行为的抑制作用。结论抑制miR-1269a可能增强非小细胞肺癌的放疗敏感性,其机制可能是通过上调SOX7的表达所致。Objective To investigate the effect of miR-1269a expression on the radiotherapy sensitivity of H460 stem celllike cells of non-small cell lung cancer.Methods Non-small cell lung cancer H460 cell line was selected for serum-free suspension culture,and CD133 and CD44 positive H460 stem cells were screened by flow cytometry.qRT-PCR was performed to detect the expression levels of miR-30a,miR-148a,miR-148b,miR-152,miR-411,miR-497,miR-598,miR-424,miR-761,miR-1269a,miR-1280 and CD44,CD133 mRNA in H460 stem cell-like cells.The effects of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell-like cells were analyzed using CCK-8 experiments,stem cell pelleting experiments,flow cytometry,and comet experiments.Results The expressions of CD133 and CD44 mRNA increased 1.30±0.03 times and 1.19±0.02 times,respectively,in H460 stem cell-like cells than in H460 cells(P<0.05).The expression level of miR-1269a was the highest in H460 stem cell-like cells,and was higher than that in H460 cells(P<0.05).After transfection of miR-1269a inhibitor and X-ray treatment,the growth inhibition rate of H460 stem cell-like cells increased significantly(72.11%±2.45%),the apoptosis rate reached to 17.70%±0.54%,and the fragmentation of DNA double-strand breaks in cells also increased significantly.After 2 Gy of X-ray radiation,the tail moment was 1.19±0.02 times of that before transfection,implying that transfection of miR-1269a inhibitor significantly increased the sensitivity of H460 stem cell-like cells to radiotherapy(P<0.05).After transfection of pcDNA3.1-SOX7,the effect of X-ray treatment on H460 stem cell-like cells was similar to that of miR-1269a inhibitor transfection.Simultaneous transfection of pcDNA3.1-SOX7 and miR-1269a mimics reversed the inhibitory effect of transfection of pcDNA3.1-SOX7 on the malignant biological behavior of H460 stem cell-like cells.Conclusion Inhibition of miR-1269a may enhance the radiotherapy sensitivity of non-small cell lung cancer,and the mechanism may be caused by up-regulation of SO

关 键 词：miR-1269a 癌 非小细胞肺 干细胞样细胞 放射治疗 放疗敏感性 

分 类 号：R734.2[医药卫生—肿瘤]