Has_circ_0065901通过竞争性吸附miR-197-3p促进 PCDH 10表达抑制宫颈癌进展的机制研究

A Study of Has_circ_0065901 Inhabitation and Mechanism for the Expression of PCDH 10 Through Competitive Adsorptionof MiR-197-3p in Cervical Cancer Progression

目的本研究以has_circ_0065901为研究对象,探讨其在宫颈癌中的分子机制。方法本研究共纳入30对取自新疆医科大学第二附属医院2019年1月至2020年12月患者,对其未经化疗或放疗的宫颈癌组织和癌旁组织进行分析。根据基因表达综合(GEO)数据库(GSE102686)分析发现了异常表达的环状RNA(circRNAs)。通过靶向预测数据库(Targetscan)发现has_circ_0065901的靶向微小RNA(miRNA)是miR-197-3p。通过TargetScan和癌症基因组图谱(TCGA)数据库发现原钙黏蛋白10(PCDH 10)是miR-197-3p的靶基因之一。此外,荧光素酶报告基因测定证实了靶标关系。采用荧光定量PCR(qRT-PCR)和免疫印迹法(WB)定量测定has_circ_0065901、miR-197-3p和PCDH 10的表达。利用细胞计数试剂盒(CCK-8)、流式细胞仪和细胞迁移检测细胞的增殖能力和迁移能力。最后通过体内裸鼠成瘤实验对其进行验证。结果发现在宫颈癌组织细胞中has_circ_0065901表达下调并能够抑制肿瘤细胞增殖和迁移,且差异具有统计学意义(P<0.05)。进一步检测发现has_circ_0065901的靶基因miR-197-3p在宫颈癌中高表达且能够促进肿瘤增殖和迁移(P<0.05)。此外,PCDH 10在宫颈癌组织及细胞中低表达,过表达PCDH 10能够显著抑制肿瘤细胞增殖和迁移且差异具有统计学意义(P<0.05)。双荧光素酶报告基因检测验证has_circ_0065901、miR-197-3p及PCDH 10三者之间的靶向关系且差异均存在统计学意义(P<0.05)。结论Has_circ_0065901能够竞争性吸附miR-197-3p,通过调节PCDH 10的表达来抑制细胞的增殖和迁移。

Objective This study attempted to explore the molecular mechanism of has_circ_0065901 in cervical cancer.Methods A total of 30 pairs of cervical cancer tissues and paracancerous tissues from patients without chemotherapy or radiotherapy from January,2019 to December,2020 in the Second Affiliated Hospital of Xinjiang Medical University were included in this study for further analysis.According to the Gene Expression Omnibus(GEO)database(GSE102686),abnormal expression of circular RNA(circRNAs)was identified.Through TargetScan database,it was found that the target miRNA of has_circ_0065901 was miR-197-3p.Using TargetScan and the Cancer Genome Atlas(TCGA)database,PCDH 10 was found to be one of the targeted genes of miR-197-3p.In addition,luciferase reporter gene assay confirmed the targeting relationship.The expressions of has_circ_0065901,miR-197-3p and PCDH 10 were quantitatively determined by fluorescent quantitative PCR(qRT-PCR)and western blotting(WB).Cell counting kit-8(CCK-8),flow cytometry and trans-well analysis method were used to analyze cell proliferation and migration ability.Finally,it was further verified by in vivo nude mouse tumor formation experiments.Results The expression of has_circ_0065901 was down-regulated in cervical cancer tissue cells and could inhibit tumor cell proliferation and migration,and the difference was statistically significant(P<0.05).Further testing found that the targeted gene miR-197-3p of has_circ_0065901 was highly expressed in cervical cancer and could promote tumor proliferation and migration(P<0.05).In addition,PCDH 10 was lowly expressed in cervical cancer tissues and cells,and overexpression of PCDH 10 could significantly inhibit tumor cell proliferation and migration with a statistically significant difference(P<0.05).The targeting relationship among has_circ_0065901,miR-197-3p and PCDH 10 through dual luciferase reporter gene detection,and the differences were statistically significant(P<0.05).Conclusion Has_circ_0065901 can competitively adsorb miR-197-3p and inhibit cell proliferation and migration by regulating the expression of PCDH 10.