沉默circCCDC66靶向miR-129-5p对食管癌细胞Eca-109增殖及凋亡的影响

Effect of Silencing circCCDC66 on the Proliferation and Apoptosis ofEsophageal Cancer Cells Eca-109 by Targeting miR-129-5p

该文探讨了沉默circCCDC66对食管癌细胞Eca-109增殖及凋亡的影响及其可能作用机制。qRT-PCR法与Western blot法分别检测食管癌组织、癌旁组织中circCCDC66、miR129-5p、HMGB1的表达量;体外培养人食管癌细胞Eca-109,将si-NC、si-circCCDC66、miR-NC、miR-129-5p mimic、si-circCCDC66+miR-129-5p inhibitor分别转染至Eca-109细胞;qRT-PCR法与Western blot法分别检测细胞中circCCDC66、miR-129-5p、HMGB1的表达量;CCK-8法、平板克隆形成实验与流式细胞术分别检测细胞增殖、集落形成数及细胞凋亡率;双荧光素酶报告实验验证circCCDC66与miR-129-5p的靶向关系,以及miR-129-5p与HMGB1的靶向关系。食管癌组织中circCCDC66、HMGB1的表达量高于癌旁组织(P<0.05),miR-129-5p的表达量低于癌旁组织(P<0.05);转染si-circCCDC66或转染miR-129-5p mimic后miR-129-5p的表达量、细胞凋亡率、细胞增殖抑制率升高(P<0.05),而HMGB1蛋白水平降低(P<0.05),集落形成数减少(P<0.05);circCCDC66可靶向调控miR-129-5p的表达,HMGB1是miR-129-5p的靶基因;共转染sicircCCDC66和miR-129-5p inhibitor可降低转染si-circCCDC66对Eca-109细胞增殖、集落形成、凋亡的影响。沉默circCCDC66可通过靶向调控miR-129-5p/HMGB1表达而降低食管癌细胞增殖、克隆形成能力,并可诱导细胞凋亡。

In this paper,the effect of silencing circCCDC66 on the proliferation and apoptosis of Eca-109in esophageal cancer cells and its possible mechanism were discussed.The expression of circCCDC66,miR-129-5p and HMGB1 in esophageal cancer tissues and adjacent tissues were detected by qRT-PCR and Western blot.Human esophageal cancer cells Eca-109 were cultures in vitro,si-NC,si-circCCDC66,miR-NC,miR-129-5pmimic,si-circCCDC66+miR-129-5p inhibitor were transfected into Eca-109 cells,respectively.The expressionof circCCDC66,miR-129-5p and HMGB1 in cells were detected by qRT-PCR and Western blot,respectively.Cell proliferation,colony formation and apoptosis rate were detected by CCK-8 assay,plate clone formation assay and flow cytometry,respectively.The targeting relationship between circCCDC66 and miR-129-5p,and thetargeting relationship between miR-129-5p and HMGB1 were verified by dual luciferase report experiment.Theexpression levels of circCCDC66 and HMGB1 in esophageal cancer tissues were higher than those in paracancertissues(P<0.05),while the expression of miR-129-5p was lower than those in paracancer tissues(P<0.05).Aftertransfection with si-circCCDC66 or miR-129-5p mimic,the expression of miR-129-5p,the rate of apoptosis,andthe inhibition rate of cell proliferation were increased(P<0.05),while the protein level of HMGB1 and the number of colonies formed were decreased(P<0.05).circCCDC66 could target and regulate the expression of miR129-5p,and HMGB1 is the target gene of miR-129-5p.Co-transfection of si-circCCDC66 and miR-129-5p inhibitor could reduce the effect of transfection of si-circCCDC66 on the proliferation,clone formation and apoptosisof Eca-109 cells.Silencing circCCDC66 can reduce the proliferation and clonal formation ability of esophagealcancer cells and induce cell apoptosis by targeting the expression of miR-129-5p/HMGB1.