首页|hsa_circ_0011946靶向miR-767-3p调控宫颈癌细胞增殖、迁移和侵袭

hsa_circ_0011946靶向miR-767-3p调控宫颈癌细胞增殖、迁移和侵袭

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目的 探讨hsa_circ_0011946靶向miR-767-3p在宫颈癌进展中的作用.方法 RT-qPCR分析hsa_circ_0011946、miR-767-3p在宫颈癌组织、癌旁正常组织、人宫颈上皮永生化细胞(H8)及宫颈癌细胞(SiHa、HeLa、Caski)中的表达水平.分别用si-NC、si-hsa_circ_0011946、miR-NC、miR-767-3p、pcDNA、pcDNA-hsa_circ_0011946、anti-miR-NC+si-hsa_circ_0011946、anti-miR-767-3p+si-hsa_circ_0011946 转染SiHa细胞,并设为si-NC组、si-hsa_circ_0011946 组、miR-NC组、miR-767-3p组、pcDNA组、pcDNA-hsa_circ_0011946组、anti-miR-NC+si-hsa_circ_0011946组、anti-miR-767-3p+si-hsa_circ_0011946组,未转染的细胞作为NC组.采用RT-qPCR检测各组SiHa细胞中hsa_circ_0011946和miR-767-3p的表达;CCK-8法检测细胞活力;平板克隆形成实验检测细胞克隆数;Transwell实验检测细胞迁移和侵袭数;Western blot检测MMP-2、MMP-9蛋白表达.circular RNA interactome预测miR-767-3p与hsa_circ_0011946的结合位点,双荧光素酶报告基因实验分析hsa_circ_0011946和miR-767-3p的靶向关系.结果 与癌旁正常组织比较,宫颈癌组织中hsa_circ_0011946表达水平显著升高(P<0.05),miR-767-3p表达水平显著降低(P<0.05).与H8细胞比较,SiHa、HeLa、Caski细胞中hsa_circ_0011946表达水平显著升高(P<0.05),miR-767-3p表达水平显著降低(P<0.05).与NC组比较,si-hsa_circ_0011946组SiHa细胞hsa_circ_0011946表达水平、吸光度(A)值、克隆数、迁移数、侵袭数及MMP-2、MMP-9蛋白表达水平显著降低/减少(P<0.05).与miR-NC组比较,miR-767-3p组SiHa细胞miR-767-3p表达水平显著升高(P<0.05),A值、克隆数、迁移数、侵袭数及MMP-2、MMP-2蛋白表达水平显著降低/减少(P<0.05).与pcDNA组比较,pcDNA-hsa_circ_0011946组SiHa细胞miR-767-3p 表达水平显著降低(P<0.05),hsa_circ_0011946 表达水平显著升高(P<0.05).共转染 miR-767-3p mimics 和WT-hsa_circ_0011946的SiHa细胞相对荧光素酶活性低于共转染miR-NC和WT-hsa_circ_0011946(P<0.05).hsa_circ_0011946与miR-767-3p直接特异性结合.与anti-miR-NC+si-hsa_circ_0011946 组比较,anti-miR-767-3p+si-hsa_circ_0011946 组SiHa细胞miR-767-3p表达水平显著降低(P<0.05),A值、克隆数、迁移数、侵袭数及MMP-2、MMP-9蛋白表达水平显著升高/增加(P<0.05).结论 干扰hsa_circ_0011946通过靶向上调miR-767-3p抑制宫颈癌细胞的增殖、迁移和侵袭.
hsa_circ_0011946 regulates the proliferation,migration and invasion of cervical cancer cells via targeting miR-767-3p
Objective To investigate the role of hsa_circ_0011946 targeting miR-767-3p in the progression of cervical cancer.Methods The expression levels of hsa_circ_0011946 and miR-767-3p in cervical cancer tissues,adjacent tissues,immortalized human cervical epithelial cells(H8)and cervical cancer cells(SiHa,HeLa,and Cashi)were determined by RT-qPCR.SiHa cells were transfected with si-NC,si-hsa_circ_0011946,miR-NC,miR-767-3p,pcDNA,pcDNA-hsa_circ_0011946,anti-miR-NC+si-hsa_circ_0011946,anti-miR-767-3p+ si-hsa_circ_0011946,respectively,as the si-NC group,the si-hsa_circ_0011946 group,the miR-NC group,the miR-767-3p group,the pcDNA group,the pcDNA-hsa_circ_0011946 group,the anti-miR-NC+si-hsa_circ_0011946 group,and the anti-miR-767-3p+si-hsa_circ_0011946 group,and the untransfected cells were used as the NC group.The expression of hsa_circ_0011946 and miR-767-3p of SiHa cells in each group was detected by RT-qPCR,the cell viability was detected by CCK-8,the number of cell clone was detected by plate clone formation assay,the cell migration and invasion numbers were detected by Transwell assay,and the protein expression of MMP-2 and MMP-9 was detected by Western blot.The binding site of miR-767-3p to hsa_circ_0011946 was predicted by circular RNA interactome,and the targeted relationship between hsa_circ_0011946 and miR-767-3p was analyzed by dual luciferase reporter assay.Results Compared with the adjacent tissues,the expression level of hsa_circ_0011946 in the cervical cancer tissues was significantly increased(P<0.05),while the expression level of miR-767-3p was significantly decreased(P<0.05).Compared with H8 cells,the expression levels of hsa_circ_0011946 in SiHa,HeLa and Caski cells were significantly increased(P<0.05),while the expression levels of miR-767-3p were signifi-cantly decreased(P<0.05).Compared with the NC group,the expression level of hsa_circ_0011946,absorbance(A)value,number of clone,number of migration,number of invasion and the protein expression levels of MMP-2 and MMP-9 were significantly decreased in SiHa cells in the miR-767-3p group(P<0.05).Compared with the miR-NC group,the expression level of miR-767-3p in SiHa cells in the miR-767-3p group was significantly increased(P<0.05),and the cell A value,number of clone,number of migration,number of invasion and the protein expression levels of MMP-2 and MMP-9 were significantly decreased(P<0.05).Compared with the pcDNA group,the expression level of miR-767-3p in SiHa cells in the pcDNA-hsa_circ_0011946 group was significantly decreased(P<0.05),and the expression level of hsa_circ_0011946 was significantly increased(P<0.05).The relative luciferase activity of SiHa cells co-transfected with miR-767-3p mimics and WT-hsa_circ_0011946 was lower than that of co-transfected with miR-NC and WT-hsa_circ_0011946(P<0.05).hsa_circ_0011946 bound to miR-767-3p directly and specifically.Compared with the anti-miR-NC+si-hsa_circ_0011946 group,the expression level of miR-767-3p in SiHa cells in the anti-miR-767-3p+si-hsa_circ_0011946 group was significantly decreased(P<0.05),and the cell A value,number of clone,number of migration,number of invasion and protein expression levels of MMP-2 and MMP-9 were significantly increased(P<0.05).Conclusion Interfering hsa_circ_0011946 can inhibit the proliferation,migration and invasion of cervical cancer cells through targeted up-regulation of miR-767-3p.

hsa_circ_0011946miR-767-3pcervical cancerproliferationmigrationinvasion

李文亚、谢程、彭莉

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长江大学附属仙桃市第一人民医院妇产科,湖北 仙桃 433000

长江大学附属仙桃市第一人民医院肾内科,湖北 仙桃 433000

hsa_circ_0011946 miR-767-3p 宫颈癌 增殖 迁移 侵袭

湖北省卫生健康委项目

WJ2021AB046

2024

局解手术学杂志
重庆市解剖学会,第三军医大学

局解手术学杂志

CSTPCD
影响因子:1.063
ISSN:1672-5042
年,卷(期):2024.33(2)
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