摘要
目的观察长链非编码RNA-linc00858和微小RNA-3182(miR-3182)在宫颈癌组织表达变化、对宫颈癌细胞株侵袭迁移的调控作用,分析宫颈癌细胞中linc00858、miR-3182的靶向关系。方法①40例宫颈癌患者,均行宫颈癌根治术,术中留取宫颈癌组织及癌旁组织,采用RT-qPCR法检测宫颈癌及癌旁组织中linc00858、miR-3182的表达,采用Spearman相关性分析法分析宫颈癌组织中linc00858和miR-3182相关性。②筛选人宫颈癌细胞株MS751为受试细胞,1~4组分别转染si-linc00858(小干扰linc00858,沉默linc00858表达)、OE-linc00858(过表达linc00858)、si-NC(小干扰阴性对照)、OE-NC(过表达阴性对照),转染48 h时采用Transwell实验观察各组细胞侵袭、迁移情况。培养48 h时采用RT-qPCR法检测各组细胞linc00858、miR-3182的表达。③分别采用RNA免疫沉淀测定实验、双荧光素酶报告实验分析MS751细胞中linc00858与miR-3182的靶向关系。结果①与癌旁组织比较,宫颈癌组织中linc00858相对表达量高、miR-3182相对表达量低(P均<0.05)。宫颈癌组织中linc00858和miR-3182的表达呈负相关(r=0.3432,P<0.05)。②与3组比较,培养48 h时1组细胞迁移数及细胞侵袭数均少;与4组比较,2组细胞迁移数及细胞侵袭数均多(P均<0.05)。与3组比较,培养48 h时1组细胞linc00858相对表达量低、miR-3182相对表达量高;与4组比较,2组细胞linc00858相对表达量高、miR-3182相对表达量低(P均<0.05)。③与加入IgG抗体者比较,加入抗AGO2抗体的MS751细胞linc00858和miR-3182沉淀RNA相对表达量高(P<0.05)。与一组比较,二组细胞荧光素酶活性低(P<0.05)。结论宫颈癌组织中linc00858高表达、miR-3182低表达。上调linc00858表达可促进MS751细胞的侵袭和迁移,下调linc00858表达可抑制MS751细胞的侵袭和迁移。宫颈癌细胞中linc00858可靶向负调控miR-3182表达。
Objective To observe the expression changes of long non-coding RNA-linc00858 and microRNA-3182(miR-3182)in cervical cancer(CC)tissues and their regulatory effects on invasion and migration of CC cell lines,and to analyze the targeting relationship between linc00858 and miR-3182 in CC cells.Methods①Forty patients with CC un⁃derwent radical cervical cancer surgery.During the surgery,CC tissues and adjacent tissues were collected and the expres⁃sion of linc00858 and miR-3182 in the CC tissues and adjacent tissues was detected by RT-qPCR.The correlation between linc00858 and miR-3182 in CC tissues was analyzed by Spearman correlation analysis.②We screened human CC cell line MS751 as the test cell line,and cells in the groups 1-4 were transfected with si-linc00858(small interference linc00858,silencing linc00858 expression),OE-linc00858(overexpression linc00858),si-NC(small interference negative control),and OE-NC(overexpression negative control),respectively.At 48 h of transfection,Transwell assay was used to observe the invasion and migration of cells in each group.RT-qPCR was used to detect the expression of linc00858 and miR-3182 in cells of each group after 48 h of cultivation.③The targeting relationship between linc00858 and miR-3182 in MS751 cells was analyzed by RNA immunoprecipitation assay and double Luciferase reporting test.Results①Compared with the adjacent tissues,the relative expression of linc00858 was higher and the relative expression of miR-3182 was lower in CC tissues(P<0.05).The expression of linc00858 and miR-3182 in CC tissue was negatively correlated(r=0.3432,P<0.05).②Compared with the group 3,the number of cell migration and cell invasion in the group 1 were less at 48 h of cultivation;compared with the group 4,the group 2 had more cell migration and cell invasion(both P<0.05).Compared with the group 3,the relative expression level of linc00858 in the group 1 was lower and miR-3182 was higher at 48 h of cultivation;compared with the group 4,the relative expression level of linc00
作者
吴颖
魏敏
冀瑞晴
王建
罗彩霞
单燕丽
WU Ying;WEI Min;JI Ruiqing;WANG Jian;LUO Caixia;SHAN Yanli(不详;Graduate School,Xuzhou Medical University,Xuzhou 221004,China)
出处
《山东医药》
CAS
2023年第18期21-25,共5页
Shandong Medical Journal
基金
新疆维吾尔自治区伊犁哈萨克自治州科技计划项目(YZ2021YD038)。
