摘要
目的:通过实验探究miR-142靶向高迁移率族蛋白1(high-mobility group box 1 protein,HMGB1)对宫颈癌(cervical cancer,CC)细胞生物学行为的影响及其潜在的作用机制。方法:采用实时荧光定量PCR(RT-PCR)和蛋白质免疫印迹法(Western Blot)检测CC组织和正常组织中miR-142和HMGB1 m RNA及蛋白表达水平,采用荧光素酶报告实验分析miR-142与HMGB1的靶向关系,CCK-8法检测CC细胞生存能力,克隆形成实验检测CC细胞增殖能力,划痕修复实验检测CC细胞迁移能力,基质胶侵袭实验检测CC细胞侵袭能力。结果:CC组miR-142 m RNA和蛋白表达水平显著低于正常组(P<0.05),HMGB1 m RNA和蛋白表达水平显著高于正常组(P<0.05),且CC癌组织中miR-142和HMGB1 m RNA和蛋白表达水平均呈显著负相关(r=-0.399,P=0.002;r=-0.429,P=0.001);miR-142与HMGB1存在靶向关系;CCK-8法实验、克隆形成实验、划痕修复实验和基质胶侵袭实验结果显示,miR-142 mimic组细胞生存、增殖、迁移和侵袭能力显著低于miR-NC组(P<0.05),miR-142 inhibitor组细胞生存、增殖、迁移和侵袭能力显著高于miR-NC组;Western Blot实验结果显示,HMGB1过表达时miR-142 mimic+plasmid组HMGB1蛋白表达水平显著高于miR-142 mimic+control plasmid组(P<0.05),显著低于miR-NC+plasmid组(P<0.05);CCK-8法实验、克隆形成实验、划痕修复实验和基质胶侵袭实验结果显示,HMGB1过表达时miR-142 mimic+plasmid组细胞生存、增殖、迁移和侵袭能力显著高于miR-142 mimic+control plasmid组(P<0.05),显著低于miR-NC+plasmid组(P<0.05)。结论:miR-142可通过靶向负调控HMGB1表达,进而抑制CC细胞生存、增殖、迁移和侵袭。
Objective:To investigate the effects of miR-142 targeting high mobility group protein 1(HMGB1)on biological behaviors of cervical cancer(CC)cells and its potential role mechanism through experiments.Methods:Real-time fluorescent quantitative PCR(RT-PCR)and Western Blot were used to detect the m RNA and protein expression levels of miR-142 and HMGB1 in CC tissues and normal tissues,and luciferase reporter assay was used to analyze the targeting relationship between miR-142 and HMGB1.CCK-8 assay was used to detect the viability of CC cells,and clone formation assay was used to detect the proliferation ability of CC cells,and scratch repair test was used to detect the migration ability of CC cells,and matrigel invasion test was used to detect the invasion ability of CC cells.Results:The m RNA and protein expression levels of miR-142 in CC group were significantly lower than those in normal group(P<0.05)while the m RNA and protein expression levels of HMGB1 were significantly higher than those in normal group(P<0.05),and the miR-142 in CC cancer tissues was significantly negatively correlated with m RNA and protein expression levels of HMGB1(r=-0.399,P=0.002;r=-0.429,P=0.001).miR-142 had a targeting relationship with HMGB1.Results of CCK-8 assay,clone formation assay,scratch repair test and matrigel invasion test showed that the cell viability,proliferation,migration and invasion in miR-142 mimic group were significantly lower than those in miR-NC group(P<0.05),and the cell viability,proliferation,migration and invasion in miR-142 inhibitor group were significantly higher than those in miR-NC group.Western Blot results showed that when HMGB1 was over-expressed,the protein expression level of HMGB1 in miR-142 mimic+plasmid group was significantly higher than that in miR-142 mimic+control plasmid group(P<0.05),and was significantly lower than that in miR-NC+plasmid group(P<0.05).Results of CCK-8 assay,clone formation assay,scratch repair test and matrigel invasion test showed that when HMGB1 was over-expressed,the cell viability,proliferation,migration and invasion in miR-142 mimic+plasmid group were significantly higher than those in miR-142 mimic+control plasmid group(P<0.05),and were significantly lower than those in miR-NC+plasmid group(P<0.05).Conclusion:miR-142 can negatively regulate the expression of HMGB1 by targeting,thereby inhibiting the viability,proliferation,migration and invasion of CC cells.
作者
朱争艳
郭静秋
陈雪梅
刘和宇
宋玉
雷磊
ZHU Zheng-yan;GUO Jing-qiu;CHEN Xue-mei;LIU He-yu;SONG Yu;LEI Lei(Department of Gynecology,Optics Valley,Tongren Hospital,Wuhan University,Wuhan,Hubei,430074,China;Department of Obstetrics and Gynecology,Hunan University of Traditional Chinese Medicine,Changsha,Hunan,410208,China)
出处
《现代生物医学进展》
CAS
2021年第18期3406-3412,共7页
Progress in Modern Biomedicine
基金
国家自然科学基金面上项目(30973771)
武汉市卫生和计划生育委员会科研项目(WZ18Q04)。
