circ_0000212靶向miR-139-5p对肝癌细胞增殖、迁移、侵袭、凋亡以及紫杉醇敏感性的影响

Circ_0000212 affects proliferation,migration,invasion,apoptosis,and paclitaxel sensitivity of liver cancer cells by targeting miR-139-5p

背景circ_0000212是一种新发现的非编码RNA,其高表达可促进结直肠癌进展.然而,circ_0000212在肝癌中的表达模式和作用仍然未知.目的探讨circ_0000212靶向miR-139-5p对肝癌细胞增殖、迁移、侵袭、凋亡以及紫杉醇敏感性的影响.方法RT-qPCR检测circ_0000212和miR-139-5p在肝癌组织、癌旁组织中的表达.Pearson相关分析确定肝癌组织中circ_0000212和miR-139-5p表达的关系.双荧光素酶报告实验验证circ_0000212和miR-139-5p靶向关系.将肝癌细胞HCC9204分为对照组、干扰circ_0000212组、干扰circ_0000212+miR-139-5p抑制物组、紫杉醇组、紫杉醇+干扰circ_0000212组、紫杉醇+干扰circ_0000212+miR-139-5p抑制物组.CCK-8法检测HCC9204细胞抑制率;集落形成实验检测HCC9204细胞集落形成数;流式细胞术检测HCC9204细胞凋亡率;Transwell实验检测HCC9204细胞迁移和侵袭数.结果与癌旁组织比较,肝癌组织中circ_0000212表达水平显著升高(P<0.05),miR-139-5p表达水平显著降低(P<0.05).肝癌组织中circ_0000212和miR-139-5p表达呈负相关关系.circ_0000212和miR-139-5p存在直接相互作用.与对照组比较,紫杉醇组HCC9204细胞circ_0000212表达水平显著降低(P<0.05),miR-139-5p表达水平显著升高(P<0.05).与对照组比较,干扰circ_0000212组、紫杉醇组HCC9204细胞克隆形成数、迁移数、侵袭数显著降低(P<0.05),抑制率、凋亡率显著升高(P<0.05).与干扰circ_0000212组比较,干扰circ_0000212+miR-139-5p抑制物组HCC9204细胞克隆形成数、迁移数、侵袭数显著升高(P<0.05),抑制率、凋亡率显著降低(P<0.05).与紫杉醇组比较,紫杉醇+干扰circ_0000212组HCC9204细胞克隆形成数、迁移数、侵袭数显著降低(P<0.05),抑制率、凋亡率显著升高(P<0.05).与紫杉醇+干扰circ_0000212组比较,紫杉醇+干扰circ_0000212+miR-139-5p抑制物组HCC9204细胞克隆形成数、迁移数、侵袭数显著升高(P<0.05),抑制率、凋亡率显著降低(P<0.05).结论干扰circ_0000212通过靶向上调miR-139-5p可抑制肝癌细胞增殖、迁移、侵袭能力,诱导细胞凋亡,并提高其对紫杉醇的敏感性.

BACKGROUND Circ_0000212 is a newly discovered non-coding RNA whose high expression promotes the progression of colorectal cancer.However,the expression patterns and roles of circ_0000212 in liver cancer remain unknown.AIM To investigate the effect of circ_0000212 targeting miR-139-5p on cell proliferation,migration,invasion,apoptosis,and paclitaxel sensitivity in liver cancer.METHODS RT-qPCR was applied to detect the expression of circ_0000212 and miR-139-5p in liver cancer tissues and adjacent tissues.Pearson correlation analysis was performed to determine the relationship between circ_0000212 and miR-139-5p expression in liver cancer tissues.Dual luciferase reporter assay was used to verify the targeting relationship between circ_0000212 and miR-139-5p.Liver cancer HCC9204 cells were divided into a control group,circ_0000212 interference group,circ_0000212 interference+miR-139-5p inhibitor group,paclitaxel group,paclitaxel+circ_0000212 interference group,and paclitaxel+circ_0000212 interference+miR-139-5p inhibitor group.The rate of inhibited HCC9204 cells was detected using CCK-8 method;the number of clones formed by HCC9204 cells was calculated using colony formation assay;the apoptotic rate of HCC9204 cells was evaluated by flow cytometry;and the migration and invasion of HCC9204 cells were detected by Transwell assay.RESULTS Compared with adjacent tissue,the expression level of circ_0000212 in liver cancer tissue was significantly increased(P<0.05),while the expression level of miR-139-5p was significantly decreased(P<0.05).There was a negative correlation between the expression of circ_0000212 and miR-139-5p in liver cancer tissues.Circ_0000212 directly interacted with miR-139-5p.Compared with the control group,circ_0000212 expression in HCC9204 cells in the paclitaxel group was significantly reduced(P<0.05),while miR-139-5p expression was significantly increased(P<0.05).Compared with the control group,the numbers of clones formed and migrating and invading HCC9204 cells in the interference circ_0000212 group and paclitaxel group were significantly reduced(P<0.05),and the inhibition rate and apoptosis rate were significantly increased(P<0.05).Compared with the circ_0000212 interference group,the numbers of clone formed and migrating and invading HCC9204 cells in the interference circ_0000212+miR-139-5p inhibitor group were significantly increased(P<0.05),and the inhibition rate and apoptosis rate were significantly reduced(P<0.05).Compared with the paclitaxel group,the numbers of clones formed and migrating and invading HCC9204 cells in the paclitaxel+circ_0000212 interference group were significantly reduced(P<0.05),and the inhibition rate and apoptosis rate were significantly increased(P<0.05).Compared with the paclitaxel+circ_0000212 interference group,the numbers of clones formed and migrating and invading HCC9204 cells in the paclitaxel+circ_0000212 interference+miR-139-5p inhibitor group were significantly increased(P<0.05),and the inhibition rate and apoptosis rate were significantly reduced(P<0.05).CONCLUSION Interfering with circ_0000212 can inhibit cell proliferation,migration,and invasion,induce cell apoptosis,and increase its sensitivity to paclitaxel in liver cancer cells by targeting and up-regulating miR-139-5p.