环状RNA同源域相互作用蛋白激酶3在三阴性乳腺癌细胞侵袭中的作用及机制  

The role of circular RNA homeodomain interacting protein kinase 3 in triple-negative breast cancer cell invasion

作　　者：吕洁 陈中华 王博[1] LU Jie;CHEN Zhonghua;WANG Bo(School of Clinical Medicine,Jiamusi University,Jiamusi 154002;Jiamusi Tumor Hospital,Jiamusi 154002,Heilongjiang,China)

机构地区：[1]佳木斯大学临床医学院,黑龙江佳木斯154002 [2]佳木斯市肿瘤医院,黑龙江佳木斯154002

出　　处：《癌变.畸变.突变》2025年第1期39-44,51,共7页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：黑龙江省卫生健康委科研课题(2020-329)。

摘　　要：目的:研究环状RNA同源域相互作用蛋白激酶3(circHIPK3)在三阴性乳腺癌(TNBC)细胞侵袭中的调控作用及机制。方法:收集2022年1月—2023年6月手术切除的TNBC组织及对应癌旁组织各40例、非三阴性乳腺癌组织及对应的癌旁组织各40例,采用荧光定量PCR法(qPCR)检测circHIPK3的表达水平。同时培养人正常乳腺上皮细胞株MCF10A,非三阴性乳腺癌细胞株MCF-7、SK-BR-3,TNBC细胞株HCC1806、MDA-MB-231、HCC-70,采用qPCR检测circHIPK3的表达水平,依据其表达水平,筛选出circHIPK3表达水平最高的MDA-MB-231细胞用于后续试验。筛选出敲低circHIPK3效果最显著的siRNA序列,然后将circHIPK3 siRNA和miR-485-3共转染MDA-MB-231细胞,采用qPCR检测circHIPK3、miR-485-3p的表达,采用Western blot法检测FGF2的蛋白表达,Transwell侵袭试验检测细胞侵袭数目。结果:乳腺癌组织中circHIPK3的相对表达水平高于对应的癌旁组织(P<0.05);TNBC组织中circHIPK3的相对表达水平高于非三阴性乳腺癌组织(P<0.05);TNBC细胞株中circHIPK3的相对表达水平高于非三阴性乳腺癌细胞株和正常乳腺癌上皮细胞株(P<0.05),且MDA-MB-231细胞中circHIPK3的表达水平高于HCC1806、HCC-70细胞(P<0.05);与转染siRNA对照序列的细胞比较,转染circHIPK3 siRNA的MDA-MB-231细胞中circHIPK3、FGF2蛋白的表达水平及细胞侵袭数目降低,miR-485-3p的表达水平升高(P<0.05);与转染miRNA对照序列的细胞比较,转染miR-485-3p抑制物的MDA-MB-231细胞(与circHIPK3 siRNA共转染)中miR-485-3p的表达水平降低,FGF2蛋白的表达水平及细胞侵袭数目增加(P<0.05)。结论:circHIPK3促进TNBC细胞侵袭,其可能的机制为通过抑制miR-485-3p的表达进而促进FGF2蛋白的表达。OBJECTIVE:To investigate the role of circular RNA homeodomain interacting protein kinase 3(circHIPK3)in triple-negative breast cancer(TNBC)cell invasion.METHODS:From January 2022 to June 2023,40 cases of surgically removed TNBC tissues,40 cases of non-triple negative breast cancer tissues and 40 cases of corresponding paracancer tissues were collected.Expression level of circHIPK3 was detected by fluorescence quantitative PCR.Normal human mammary epithelial cell lines MCF10A,non-triple negative breast cancer cell lines MCF-7 and SK-BR-3,TNBC cell lines HCC1806,MDA-MB-231 and HCC-70 were cultured.,Their expression levels of circHIPK3 were detected by fluorescence quantitative PCR.According to the expression level,MDA-MB-231 cells with the highest circHIPK3 expression were selected for grouping.The siRNA sequence with the most significant circHIPK3 knockdown effect was screened,and then circHIPK3 siRNA and miR-485-3 were co-transfected into MDA-MB-231 cells.Expressions of circHIPK3 and miR-485-3p were detected by fluorescence quantitative PCR.Protein expression of FGF2 was detected by western blot.The number of cell invasions was detected by the transwell invasion assay.RESULTS:The relative expression levels of circHIPK3 in breast cancer tissues were higher than that in adjacent tissues(P<0.05).The relative expression level of circHIPK3 in TNBC tissues was higher than that in non-triple negative breast cancer tissues(P<0.05).The relative expression level of circHIPK3 in TNBC cell lines was higher than that in non-triple negative breast cancer cell lines and normal breast cancer epithelial cell lines(P<0.05),and the expression level of circHIPK3 in MDA-MB-231 cells was higher than that in HCC1806 and HCC-70 cells(P<0.05).Compared with the cells transfected with control siRNA sequence,the expression levels of circHIPK3 and FGF2 and the number of cell invasion in MDA-MB-231 cells transfected with circHIPK3 siRNA were decreased,and the expression level of miR-485-3p was increased(P<0.05).Compared with cells transfected

关 键 词：三阴性乳腺癌 环状RNA同源域相互作用蛋白激酶3 成纤维细胞生长因子2 细胞侵袭 

分 类 号：R737.9[医药卫生—肿瘤]