宫颈癌细胞中ΔNp63α的MicroRNA表达谱及其对hsa-let-7b-3p的调控机制  

MicroRNA Expression Profile of ΔNp63α in Cervical Cancer Cells and Its Regulation Mechanism on hsa-let-7b-3p

作　　者：宋伟国 钱莉莉 吴大保 申震 朱靖 程勇 肖卫华 周颖 SONG Weiguo;QIAN Lili;WU Dabao;SHEN Zhen;ZHU Jing;CHENG Yong;XIAO Weihua;ZHOU Ying(Department of Obstetrics and Gynecology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, China;Department of Radiation, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, China;The CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Life Sciences, University of Science and Technology Of China, Hefei 230021, China)

机构地区：[1]安徽医科大学附属省立医院妇产科,合肥230001 [2]安徽医科大学附属省立医院放疗科,合肥230001 [3]中国科学院天然免疫与慢性疾病重点实验室 中国科学技术大学生命科学院,合肥230021

出　　处：《肿瘤防治研究》2018年第3期138-143,共6页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金(81272881;81001168);安徽省自然科学基金(1308085MH122);安徽省科技攻关项目(1501041141);中国科学院天然免疫与慢性疾病重点实验室开放课题(KLIICD-201603)

摘　　要：目的在宫颈鳞癌细胞中筛选出ΔNp63α显著调控的mi RNAs,并探讨其对宫颈癌细胞功能的影响。方法通过mi RNA芯片技术筛选出ΔNp63α过表达的Si Ha细胞株(Si Ha/p63)中差异明显的mi RNAs,并进行q RT-PCR验证;通过q RT-PCR在沉默ΔNp63α的ME-180细胞株(ME-180/Shp63)中检测差异表达的mi RNAs,并选择差异较显著的hsa-let-7b-3p作为研究对象;向Si Ha细胞中转染hsa-let-7b-3p刺激剂,通过生长曲线、Western blot以及流式细胞仪检测hsa-let-7b-3p对Si Ha细胞增殖及凋亡的影响。结果 (1)在ΔNp63α过表达的Si Ha细胞株中,筛选出10个变化较显著的mi RNAs;(2)在ΔNp63α沉默的ME-180细胞中,筛选出5个与上述变化趋势一致的mi RNAs;(3)hsa-let-7b-3p过表达抑制Si Ha细胞的增殖;(4)hsa-let-7b-3p过表达促进Si Ha细胞的凋亡。结论宫颈癌细胞株中,mi RNA hsalet-7b-3p的表达与ΔNp63α的表达呈正相关,且可以有效抑制细胞增殖,其作用机制可能与诱导细胞周期阻滞和促进细胞凋亡相关。Objective To screen out the mi RNAs regulated by ΔNp63α in squamous cell carcinoma of the cervix and investigate their effect on the function of cervical cancer cells. Methods mi RNA microarray was used to screen the mi RNAs with significant difference in Si Ha cells with overexpressed ΔNp63α and verified by q RT-PCR. The mi RNAs with differential expression were validated by q RT-PCR in ME-180 cells with silenced ΔNp63α and the hsa-let-7 b-3 p was selected as the study object. The effect of hsa-let-7 b-3 p on the proliferation and apoptosis of Si Ha cells was detected by growth curve, Western blot and flow cytometry after the transfection of hsa-let-7 b-3 p mimic into Si Ha cells. Results（1） In Si Ha cells with ΔNp63α overexpression, ten mi RNAs consistent with the trend of q RT-PCR were screened out from the mi RNAs with significant differences in the results of the microarray.（2） Five mi RNAs that were consistent with the above change trends were screened out in ME-180 cells with silenced ΔNp63α.（3） Overexpression of hsa-let-7 b-3 p inhibited the proliferation of Si Ha cells.（4） Overexpression of hsa-let-7 b-3 p promoted the apoptosis of Si Ha cells. Conclusion In the cervical cancer cell line, the expression of mi RNA hsa-let-7 b-3 p is positively correlated with the expression of ΔNp63α, and it can inhibit cell proliferation effectively. The mechanism may be related to the induction of cell cycle arrest and the promotion of apoptosis.

关 键 词：ΔNp63α 子宫颈癌 miRNA hsa-let-7b-3p 增殖 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]