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

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Si Ha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,Si Ha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。

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,GADD45 B 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 RTPCR after chemotherapy in Si Ha 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.