Circ_DOCK1调节miR-122-5p/PPIB轴对结直肠癌细胞增殖、迁移和侵袭的影响

Effects of circ_DOCK1 on the proliferation,migration and invasion of colorectal cancer cells by regulating the miR-122-5p/PPIB axis

目的探讨环状核糖核苷酸_胞质分裂作用因子1(Circ_DOCK1)通过调节微小核糖核苷酸-122-5p(miR-122-5p)/肽酰脯氨基顺反异构酶B(PPIB)轴对结直肠癌细胞增殖、迁移和侵袭的影响。方法实时荧光定量PCR检测结直肠癌患者癌组织与癌旁组织中的circ_DOCK1、miR-122-5p、PPIB表达水平。取人结直肠细胞SW480,脂质体转染法转染circ_DOCK1干扰质粒(sh-circ_DOCK1)、miR-122-5p抑制物(anti-miR-122-5p)和模拟物(miR-122-5p mimics),以CCK-8法、划痕试验和Transwell实验检测结直肠癌细胞的增殖、迁移和凋亡情况。以生物信息学和荧光素酶报告基因实验分析circ_DOCK1与miR-122-5p的靶向关系。结果结直肠癌组织的circ_DOCK1、PPIB mRNA表达水平高于癌旁组织,miR-122-5p表达水平低于癌旁组织。与挽救组和NC-circ_DOCK1组比较,sh-circ_DOCK1组的circ_DOCK1和PPIB mRNA表达量下降,miR-122-5p表达量升高,且细胞转染48 h和72 h的A值及细胞迁移率和穿膜细胞数均降低(P<0.05);挽救组和NC-circ_DOCK1组的数据比较差异无统计学意义(P>0.05)。生物信息学软件预测circ_DOCK1含有与miR-122-5p互补的核苷酸序列。转染miR-122-5p mimics后,circ_DOCK1的野生型荧光素酶报告质粒相对活性降低,转染anti-miR-122-5p后,circ_DOCK1的野生型荧光素酶报告质粒相对转录活性增加(P<0.05),circ_DOCK1突变型荧光素酶报告质粒相对活性值变化无统计学意义(P>0.05)。结论circ_DOCK1在结肠癌组织中的表达上调,且能通过调节miR-122-5p/PPIB轴影响结直肠癌的增殖、迁移和侵袭,可考虑经circ_DOCK1作为结直肠癌的分子靶向治疗研究方向。

Objective To investigate the effects of Circ_DOCK1 on the proliferation,migration and invasion of colorectal cancer cells by regulating microribonucleotide-122-5p(miR-122-5p)/peptidylproamino cis-trans isomerase B(PPIB)axis.Methods The expression levels of circ_DOCK1,miR-122-5p and PPIB in cancer tissues and adjacent tissues of colorectal cancer patients were detected by real-time fluorescence quantitative PCR.Human colorectal cells SW480 were transfected with circ_DOCK1 interfering plasmid(sh-circ_DOCK1),miR-122-5p inhibitor(anti-miR-122-5p)and miR-122-5p mimics(miR-122-5p mimics).The proliferation,migration and apoptosis of colorectal cancer cells were detected by CCK-8 method,scratch test and Transwell test.The targeting relationship between circ_DOCK1 and miR-122-5p was analyzed by bioinformatics and luciferase reporter gene experiments.Results The expression levels of circ_DOCK1 and PPIB mRNA in colorectal cancer tissues were higher than those in adjacent tissues,and the expression level of miR-122-5p was lower than that in adjacent tissues.Compared with the rescue group and NC-circ_DOCK1 group,the expression of circ_DOCK1 and PPIB mRNA in sh-circ_DOCK1 group decreased,expression of miR-122-5p increased,and the A value,cell migration rate and number of transmembrane cells at 48 h and 72 h after transfection decreased(P<0.05),which were no significant difference between rescue group and NC-circ_DOCK1 group(P>0.05).Bioinformatics software predicted that circ_DOCK1 contained a nucleotide sequence complementary to miR-122-5p.The dual luciferase reporter gene experiment showed that the relative activity of the wild-type luciferase reporter plasmid of circ_DOCK1 decreased after transfection of miR-122-5p mimics,and the wild-type luciferase of circ_DOCK1 after transfection of anti-miR-122-5p The relative transcriptional activity of the reporter plasmid increased(P<0.05).Conclusion The expression of circ_DOCK1 is up-regulated in colon cancer tissue,and it can affect the proliferation,migration and invasion of colorectal cancer by regulating the miR-122-5p/PPIB axis.Circ_DOCK1 can be considered as the research direction of molecular targeted therapy for colorectal cancer.