miR-29b-2-5p通过ABCC1抑制结直肠癌恶性生物学行为的作用机制

The Mechanism by Which miR-29b-2-5p Inhibits the Malignant Biological Behavior of Colorectal Cancer by Targeting ABCC1

目的:探讨miR-29b-2-5p在调节结直肠癌(CRC)恶性生物学行为过程中的分子机制。方法:通过qRT-PCR技术分析CRC组织和细胞系(HCT-116、HT-29、Caco2、SW480和NCM460)中miR-29b-2-5p和ABCC1的转录组表达水平。分别使用细胞计数和Transwell进行评估HT-29细胞的增殖和侵袭能力。使用双荧光素报告系统验证miR-29b-2-5p与ABCC1之间的靶向作用。结果:CRC组织和细胞系中miR-29b-2-5p的表达水平显著低于正常组织(P<0.001),尤其在HT-29细胞系中miR-29b-2-5p的表达水平最为显著降低。过表达miR-29b-2-5p或沉默ABCC1均显著抑制HT-29细胞增殖(P<0.05)与侵袭(P<0.01)。生物信息学预测结果显示,ABCC1的3’UTR区域含有miR-29b-2-5p结合位点。进一步实验表明,miR-29b-2-5p过表达导致ABCC1转录表达水平显著下调(P<0.001)。双荧光素酶报告系统检测结合TCGA公共数据库结果显示,miR-29b-2-5p直接靶向ABCC1 3’UTR区域(P<0.001),并且二者表达水平呈显著负相关(P<0.001)。结论:miR-29b-2-5p通过靶向ABCC1的方式参与调控HT-29细胞的增殖和侵袭,参与CRC的发生发展。

Objective:To investigate the molecular mechanism of miR-29b-2-5p in regulating the malignant biological behavior of colorectal cancer(CRC).Methods:The transcriptional expression levels of miR-29b-2-5p and ABCC1 in CRC tissues and cell lines(HCT-116,HT-29,Caco2,SW480 and NCM460)were analyzed using qRT-PCR assay.Cell proliferation and invasion abilities of HT-29 cells were evaluated using cell counting and Transwell assays,respectively.The targeting effect of miR-29b-2-5p on ABCC1 was validated using a dual-luciferase reporter system.Results:The expression level of miR-29b-2-5p in CRC tissues and cell lines was significantly lower than in normal tissues(P<0.001),with the most significant decrease observed in HT-29 cell line.Overexpression of miR-29b-2-5p or silencing of ABCC1 significantly inhibited HT-29 cell proliferation(P<0.05)and invasion(P<0.01).Bioinformatics predictions indicated that the 3’UTR region of ABCC1 contained binding sites for miR-29b-2-5p.Further experiments demonstrated that overexpression of miR-29b-2-5p led to a significant downregulation of ABCC1 transcriptional expression levels(P<0.001).Dual-luciferase reporter system analysis Combined with TCGA confirmed direct targeting of the ABCC13’UTR region by miR-29b-2-5p(P<0.001),with a significant negative correlation between their expression levels(P<0.001).Conclusion:miR-29b-2-5p participates in regulating the proliferation and invasion of HT-29 cells by targeting ABCC1,thereby contributing to the occurrence and development of CRC.