miR-603对宫颈癌SiHa细胞增殖和迁移的影响及其机制  

Effect of miR-603 on proliferation and migration of cervical cancer cells SiHa and its mechanism

作　　者：芦娇娇 甄帅[2] 李旭[2] LU Jiaojiao;ZHEN Shuai;LI Xu(Department of Radiology,First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China;Center for Translational Medicine,First Affiliated Hospital of Xi′an Jiaotong University)

机构地区：[1]西安交通大学第一附属医院医学影像科,西安710061 [2]西安交通大学第一附属医院转化医学中心

出　　处：《山西医科大学学报》2024年第4期439-446,共8页Journal of Shanxi Medical University

基　　金：西安交通大学第一附属医院科研发展基金项目(2020QN-27)。

摘　　要：目的探讨miR-603抑制宫颈癌细胞增殖和迁移的机制。方法采用基因编辑系统(clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9,CRISPR/Cas9)分别敲除宫颈癌细胞SiHa中人乳头瘤病毒16(human papilloma virus 16,HPV16)E6、E7后采用real-time PCR检测miR-603的表达。将宫颈癌细胞SiHa分为4组:mimic NC组、mimic 603组、inhibitor NC组和inhibitor 603组,将mimic NC、mimic 603、inhibitor NC及inhibitor 603分别转染SiHa细胞,通过real-time PCR检测miR-603的表达水平,CCK-8法检测SiHa细胞增殖能力,Transwell小室法检测SiHa细胞迁移能力。双荧光素酶报告试验检测miR-603是否与IQ结构域GTP酶激活蛋白3(IQ motif-containing GTPase activating protein 3,IQGAP3)直接结合,Western blotting检测IQGAP3蛋白表达。UALCAN(The University of Alabama at Birmingham cancer data analysis portal)数据库分析IQGAP3在肿瘤中的表达情况;通过肿瘤细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库分析IQGAP3在宫颈癌细胞系中的表达;利用分析癌症基因表达和病毒感染的交互式在线数据库(an interactive online database for analysis of gene expression and viral infection in cancer,OncoDB)分析IQGAP3与宫颈癌患者HPV感染的相关性。结果敲除HPV16 E6、E7后miR-603的表达升高(P<0.05)。过表达miR-603后,SiHa细胞的增殖和迁移能力下降(P<0.05);敲低miR-603后,SiHa细胞的增殖和迁移能力增强(P<0.05)。双荧光素酶报告试验提示IQGAP3是miR-603的直接靶点,过表达miR-603后,IQGAP3的表达降低,敲低miR-603可促进IQGAP3的表达。UALCAN数据库显示IQGAP3在多种肿瘤组织中表达高于正常对照,OncoDB数据库分析发现HPV16、18阳性患者IQGAP3表达均高于阴性患者。使用CCLE数据库分析IQGAP3在宫颈癌细胞系的表达,发现HPV阴性C33A细胞中IQGAP3表达明显低于HPV16阳性CASKI、SiHa细胞,以及HPV18阳性HELA、C4I、C4II及MS751细胞。结论敲除HPV16 E6、E7后miR-603�Objective To explore the mechanism of miR-603 inhibiting the proliferation and migration of cervical cancer cells.Methods The clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9(CRISPR/Cas9)gene editing system was used to knock out human papillomavirus 16(HPV16)E6 and E7 in SiHa cells,respectively,and then miR-603 expression was detected by real-time PCR.Cervical cancer SiHa cells were divided into four groups:mimic NC group,mimic 603 group,inhibitor NC group and inhibitor 603 group,and transfected with mimic NC,mimic 603,inhibitor NC,and inhibitor 603,respectively.The miR-603 expression level was detected by real-time PCR,the cell proliferation capacity of SiHa cells was assessed by CCK-8 assay,and the cell migration ability was evaluated using Transwell assay.Dual luciferase reporter assay was used to detect whether miR-603 directly binds to IQ motif-containing GTPase activating protein 3(IQGAP3),and IQGAP3 protein expression was detected by Western blotting.The University of Alabama at Birmingham cancer data analysis portal(UALCAN)database was utilized to analyze the expression of IQGAP3 in multiple tumors.The Cancer Cell Line Encyclopedia(CCLE)database was used to analyze the expression of IQGAP3 in cervical cancer cell lines.Additionally,an interactive online database for analysis of gene expression and viral infection in cancer(OncoDB)database was used to analyze the relationship between IQGAP3 and HPV status of cervical cancer patients.Results After knocking out HPV16 E6 or E7,the expression of miR-603 was increased(P<0.05).Overexpression of miR-603 decreased the proli-feration and migration of SiHa cells(P<0.05),while knockdown of miR-603 enhanced the proliferation and migration of SiHa cells(P<0.05).Dual luciferase reporter assay suggested that IQGAP3 was a direct target of miR-603.After overexpression of miR-603,the expression of IQGAP3 was decreased,whereas knockdown of miR-603 promoted the expression of IQGAP3.UALCAN database showed that the expression of IQGAP3 was

关 键 词：宫颈癌 miR-603 IQGAP3 细胞增殖 细胞迁移 

分 类 号：R737.33[医药卫生—肿瘤]