MiR-886-5p对宫颈鳞状上皮细胞克隆形成和宫颈癌细胞化学药物治疗的影响

doi:10.3969/j.issn.1006-7795.2015.06.017

首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (6): 929-935.doi: 10.3969/j.issn.1006-7795.2015.06.017

MiR-886-5p对宫颈鳞状上皮细胞克隆形成和宫颈癌细胞化学药物治疗的影响

李晶华¹, 王明², 张瑞¹, 冯力民¹

1. 首都医科大学附属北京天坛医院妇产科, 北京 100050;
2. 首都医科大学附属北京佑安医院妇产科, 北京 100069

收稿日期:2015-07-07 出版日期:2015-12-21 发布日期:2015-12-18
通讯作者: 冯力民 E-mail:lucyfeng1966@163.com
基金资助:
国家自然科学基金(81101969)。

Effect of the miR-886-5p on the cervical squamous epithelial cell clone formation and cervical cancer chemotherapy

Li Jinghua¹, Wang Ming², Zhang Rui¹, Feng Limin¹

1. Department of Obstetrics and Gynecology, Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China;
2. Department of Obstetrics and Gynecology, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China

Received:2015-07-07 Online:2015-12-21 Published:2015-12-18
Supported by:
This study was supported by National Natural Science Foundation of China(81101969).

摘要/Abstract

摘要： 目的探讨microRNA-886-5p(MiR-886-5p)对宫颈鳞状上皮细胞克隆形成及宫颈癌细胞化学药物治疗(以下简称化疗)的影响。方法应用基因芯片技术检测宫颈癌及其癌周组织microRNA的表达,筛选出MiR-886-5p在宫颈癌组织中高表达。生物信息系技术分析发现miRNA-886-5p靶向P53通路中多个基因的表达。用MiR-886-5p mimics和带有GFP标签的MiR-886-5p过表达载体稳定转染高危型人乳头瘤病毒(human papilomavirus,HPV16)阳性的永生化人宫颈鳞状上皮细胞H8细胞,应用Western blotting方法检测P53和P14的蛋白表达情况;应用平板克隆形成技术,观察对细胞克隆形成的影响。在宫颈癌SiHa细胞中,加入化疗药物紫杉醇和VP16后,应用real time RT-PCR技术,检测MiR-886-5p的表达情况。结果过表达 MiR-886-5p后,H8细胞的克隆形成率明显高于阴性对照组,并且降调P53通路中P14和P53蛋白的表达。加入化疗药紫杉醇和VP16后,SiHa细胞中MiR-886-5p表达明显升高。结论 MiR-886-5p与宫颈鳞状细胞增生相关,它降调P14和P53两种蛋白的表达,且与宫颈癌细胞化疗抵抗相关。

关键词: 宫颈鳞状上皮细胞, 宫颈癌, MiR-886-5p, 克隆形成, 化学治疗

Abstract: Objective To explore the effect of the miR-886-5p on the cervical squamous epithelial cell clone formation and cervical cancer chemotherapy. Methods Microassay was carried out for cervical squamous cell carcinoma tissues(CSCCs) and adjacent non-tumor tissues. One of them is miR-886-5p that overexpression in CSCCs. Bioinformatics analysis suggests that P53 pathway genes(Bax, P14, GADD45B and 14-3-3δ) are miR-886-5p target genes. With a GFP tag overexpression of miR-886-5p plasmid we evaluated the formation of cervical epithelial cell clone. Downstream target validation was performed for miR-886-5p. MiR-886-5p was validated by RT-PCR after chemotherapy in SiHa cell. Results Forced expression of one miRNA, miR-886-5p, overexpressed in CSCC tissues lowered expression of the protein P14 and P53, promoted cell clone formation in H8, an HPV16-immortalized human cervical squamous epithelial cell line. After the cervical squamous carcinoma cell line was cultured with different amounts of paclitaxel and VP16, the expression of miR-886-5p was significantly upregulated with the increasing concentration of paclitaxel and VP16. Conclusion MiR-886-5p promotes proliferation of cervical cancer cells and contributes to cervical cancer chemotherapy resistance.

Key words: cervical squamous epithelial cell, cervical cancer cell, MiR-886-5p, clone formation, chemotherapy

中图分类号:

R711.74

李晶华, 王明, 张瑞, 冯力民. MiR-886-5p对宫颈鳞状上皮细胞克隆形成和宫颈癌细胞化学药物治疗的影响[J]. 首都医科大学学报, 2015, 36(6): 929-935.

Li Jinghua, Wang Ming, Zhang Rui, Feng Limin. Effect of the miR-886-5p on the cervical squamous epithelial cell clone formation and cervical cancer chemotherapy[J]. Journal of Capital Medical University, 2015, 36(6): 929-935.

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MiR-886-5p对宫颈鳞状上皮细胞克隆形成和宫颈癌细胞化学药物治疗的影响

Effect of the miR-886-5p on the cervical squamous epithelial cell clone formation and cervical cancer chemotherapy

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