circGFRA1靶向miR-3194-3p调控结直肠癌细胞增殖、迁移及侵袭OA
circGFRA1 regulates the proliferation,migration and invasion of colorectal cancer cells by targeting miR-3194-3p
目的:探讨circGFRA1对结直肠癌细胞SW620增殖、迁移及侵袭的影响及其作用机制.方法:采用qRT-PCR检测结直肠癌组织与癌旁组织、结直肠癌细胞SW620与人正常结肠上皮细胞NCM460中circGFRA1和miR-3194-3p的表达水平;体外培养SW620细胞并分为6组:si-NC组、si-circGFRA1组、miR-NC组、miR-3194-3p组、si-circGFRA1+anti-miR-NC 组、si-circGFRA1+anti-miR-3194-3p 组(si-circGFRA1+anti-miR 组).应用 CCK-8 法、平板克隆形成实验检测细胞增殖能力;划痕实验、Transwell实验检测细胞迁移及侵袭能力;双荧光素酶报告实验验证circGFRA1与miR-3194-3p的靶向关系;蛋白免疫印迹法检测MMP-2、MMP-9的蛋白表达量.结果:结直肠癌组织中circGFRA1表达水平显著高于癌旁组织(P＜0.001),miR-3194-3p表达水平显著低于癌旁组织(P＜0.001);SW620细胞中circGFRA1表达水平显著高于NCM460细胞(P＜0.001),miR-3194-3p表达水平显著低于NCM460细胞(P＜0.001).与si-NC组相比,si-circGFRA1组细胞活力、克隆形成数、迁移及侵袭细胞数均显著降低(P＜0.05),MMP-2、MMP-9蛋白表达量显著下调(P＜0.05);与miR-NC组相比,miR-3194-3p组上述指标变化趋势与si-circGFRA1组一致(P＜0.05).双荧光素酶报告实验证实circGFRA1可靶向结合miR-3194-3p.与si-circGFRA1+anti-miR-NC组相比,si-circGFRA1+anti-miR组细胞活力、克隆形成数、迁移及侵袭细胞数均显著升高(P＜0.05),MMP-2、MMP-9蛋白表达量显著上调(P＜0.05).结论:circGFRA1在结直肠癌中高表达,通过靶向抑制miR-3194-3p表达,上调MMP-2、MMP-9蛋白水平,从而促进结直肠癌细胞增殖、迁移及侵袭.
Objective:To explore the effect and mechanism of circGFRA1 on the proliferation,migration and invasion of colorectal cancer(CRC)SW620 cells.Methods:qRT-PCR was used to detect the expression levels of circGFRA1 and miR-3194-3p in CRC tissues,adjacent normal tissues,SW620 cells and normal human colonic epithelial NCM460 cells.SW620 cells were cultured in vitro and divided into 6 groups:si-NC group,si-circGFRA1 group,miR-NC group,miR-3194-3p group,si-circGFRA1+anti-miR-NC group,and si-circGFRA1+anti-miR-3194-3p(si-circGFRA1+anti-miR)group.CCK-8 assay and colony formation assay were used to detect cell proliferation ability;scratch assay and Transwell assay were used to detect cell migration and invasion abilities;dual luciferase reporter assay was used to verify the targeting relationship between circGFRA1 and miR-3194-3p;Western blotting was used to detect the protein expression levels of MMP-2 and MMP-9.Results:The expression level of circGFRA1 in CRC tissues was significantly higher than that in adjacent normal tissues(P＜0.001),and the expression level of miR-3194-3p was significantly lower than that in adjacent normal tissues(P＜0.001);the expression level of circGFRA1 in SW620 cells was significantly higher than that in NCM460 cells(P＜0.001),and the expression level of miR-3194-3p was significantly lower than that in NCM460 cells(P＜0.001).Compared with the si-NC group,the cell viability,number of colony formation,migration and invasion cells in the si-circGFRA1 group were significantly reduced(P＜0.05),and the protein expression levels of MMP-2 and MMP-9 were significantly downregulated(P＜0.05);compared with the miR-NC group,the change trends of the above indicators in the miR-3194-3p group were consistent with those in the si-circGFRA1 group(P＜0.05).Dual luciferase reporter assay confirmed that circGFRA1 could specifically bind to miR-3194-3p.Compared with the si-circGFRA1+anti-miR-NC group,the cell viability,number of colony formation,migration and invasion cells in the si-circGFRA1+anti-miR group were significantly increased(P＜0.05),and the protein expression levels of MMP-2 and MMP-9 were significantly upregulated(P＜0.05).Conclusion:circGFRA1 is highly expressed in CRC,and promotes the proliferation,migration and invasion of CRC cells by targeting and inhibiting the expression of miR-3194-3p and upregulating the protein levels of MMP-2 and MMP-9.
张荣菊;王汝月;苗志刚;孙荣国;苗玉;周玮玮
沧州市中心医院 病理科,河北沧州 061600沧州市中心医院 科研处,河北沧州 061600沧州市中心医院 病理科,河北沧州 061600沧州市中心医院 心血管内科,河北沧州 061600沧州市中心医院 病理科,河北沧州 061600沧州市中心医院 病理科,河北沧州 061600
医药卫生
结直肠癌circGFRA1miR-3194-3p增殖迁移侵袭基质金属蛋白酶
colorectal cancercircGFRA1miR-3194-3pproliferationmigrationinvasionmatrix metalloproteinase
《江苏大学学报（医学版）》 2026 (2)
136-141,6
河北省医学科学研究课题计划项目(20240590)
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