miR-4677-3p通过靶向调控KLF7影响结直肠癌细胞增殖、凋亡和侵袭OA
miR-4677-3p regulates the proliferation,apoptosis and invasion of colorectal cancer cells by targeting and modulating KLF7
目的 探讨miR-4677-3p通过靶向KLF7对结直肠癌(CRC)细胞增殖、凋亡和侵袭的影响.方法 采用Western blotting及实时PCR检测CRC组织及细胞中miR-4677-3p、KLF7表达水平.将HCT116细胞分为miR-NC组、miR-4677-3p mimics组、si-NC组、si-KLF7组、miR-4677-3p mimics+pcDNA3.1组、miR-4677-3p mimics+pcDNA-KLF7组,用集落形成实验、流式细胞术、Transwell实验分别检测细胞增殖、凋亡、侵袭情况;用Western blotting检测增殖、凋亡、侵袭相关蛋白表达.通过裸鼠移植瘤实验观察移植瘤生长情况,用实时PCR检测移植瘤组织miR-4677-3p表达水平,用免疫组织化学染色检测移植瘤组织中增殖、凋亡、侵袭相关蛋白表达.用starBase网站预测及双萤光素酶报告基因实验检测miR-4677-3p与KLF7的靶向关系.结果 在CRC组织及CRC细胞系HCT116、SW480、HT29中,miR-4677-3p表达下调,KLF7表达上调(P<0.05);过表达miR-4677-3p及沉默KLF7均可降低细胞克隆形成数、细胞侵袭数及KLF7、Ki-67、MMP-9、CLDN1表达,增高细胞凋亡率及cleaved caspase-3表达(P<0.05);而在过表达miR-4677-3p基础上沉默KLF7可部分逆转过表达miR-4677-3p对CRC细胞生长的抑制作用;miR-4677-3p与KLF7之间存在靶向关系;miR-4677-3p mimics组较miR-NC组移植瘤体积、重量减小,miR-4677-3p及cleaved caspase-3表达水平升高,KLF7、Ki-67、MMP-9、CLDN1表达水平降低(P<0.05).结论 miR-4677-3p可通过靶向抑制KLF7而抑制CRC细胞增殖、侵袭,促进其凋亡.
Objective To explore the effects of miR-4677-3p on the proliferation,apoptosis,and invasion of colorectal cancer(CRC)cells by targeting KLF7.Methods Real-time PCR and Western blotting were used to detect miR-4677-3p and KLF7 expression in CRC tissues and cells.HCT116 cells were assigned into miR-NC,miR-4677-3p mimics,si-NC,si-KLF7,miR-4677-3p mimics+pcDNA3.1,and miR-4677-3p mimics+pcDNA-KLF7 groups.Cell cloning experiments,flow cytometry,and Transwell assay were used to detect cell proliferation,apoptosis,and invasion,respectively.Western blotting was used to test proliferation,apoptosis,and invasion-related pro-teins.The nude mouse transplant tumor experiment was used to observe the growth of the transplant tumor.Real-time PCR was performed to detect miR-4677-3p in tumor tissue.Immunohistochemistry was performed to detect the proteins related to tumor tissue proliferation,apoptosis,and invasion.Results In the CRC tissues and CRC cells HCT116,SW480,and HT29,miR-4677-3p was downregulated and KLF7 was upregulated(P<0.05).The miR-4677-3p mimic group showed a decrease in the number of cell clones formed,the number of cell invasions,KLF7,Ki-67,MMP-9,and CLDN1,and an increase in the apoptosis rate and cleaved caspase-3 when compared to the miR-NC group(P<0.05).For the si-NC group,the si-KLF7 group showed a decrease in the number of cell clones formed,the number of cell invasions,Ki-67,MMP-9,and CLDN1,and an increase in the apoptosis rate and cleaved caspase-3 when compared to the si-NC group(P<0.05).The miR-4677-3p mimics+pcDNA-KLF7 group showed an increase in the number of cell clones formed,the number of cell invasions,KLF7,Ki-67,MMP-9,and CLDN1,and a decrease in the apoptosis rate and cleaved caspase-3 compared to the miR-4677-3p mimics+pcDNA3.1 group(P<0.05).The starBase website prediction and dual-luciferase assay reports indicated a targeted relationship between miR-4677-3p and KLF7.In the miR-NC group,the miR-4677-3p mimics group showed a decrease in regard to the tumor volume and mass,an increase in miR-4677-3p and cleaved caspase-3 levels,and a decrease in KLF7,Ki-67,MMP-9,and CLDN1(P<0.05).Conclusion miR-4677-3p can inhibit CRC cell proliferation,invasion,and promote apoptosis by targeting KLF7.
康嘉敏;高紫玉;刘妍慧;乌日罕;刘彩霞;邢智伟
内蒙古医科大学 研究生院,呼和浩特 010050内蒙古医科大学 附属医院肿瘤内科,呼和浩特 010050内蒙古医科大学 研究生院,呼和浩特 010050内蒙古医科大学 附属医院肿瘤内科,呼和浩特 010050内蒙古医科大学 附属医院肿瘤内科,呼和浩特 010050内蒙古医科大学 附属医院肿瘤内科,呼和浩特 010050
医药卫生
miR-4677-3pKLF7结直肠癌增殖凋亡侵袭
miR-4677-3pKLF7colorectal cancerproliferationapoptosisinvasion
《中国医科大学学报》 2026 (8)
700-706,7
内蒙古自治区"草原英才"工程创新创业团队项目(ZY20241207)
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