Zhang Yanling, Da Yang Guang, Chen Qian Ya, Zeng Jinlong, Cao Yang
Department of Oncology, The Fourth Affiliated Hospital of Guangzhou Medical University, No. 232, Outer Ring East Road, Panyu District, Guangzhou 510000 Guangdong Province, China.
The First Clinical School of Guangzhou University of Chinese Medicine, No. 1 Guangming East Road, Zengcheng District, Guangzhou 510000 Guangdong Province, China.
Toxicol Res (Camb). 2024 Dec 17;13(6):tfae149. doi: 10.1093/toxres/tfae149. eCollection 2024 Dec.
To investigate the role and mechanism of miR-342 and FOXP1 on hepatocellular carcinoma cells. QRT-PCR was applied to determine the expression of miR-342, FOXP1 and MYCBP in normal hepatocyte cell lines (NHC), hepatocellular carcinoma cell lines (HEK-293 T) and human hepatocellular carcinoma cell lines (HepG2, MHCC97-L, Huh7 and SMMC7721). After knockdown or over-expression of miR-342 and FOXP1 in HepG2 cells respectively, cell proliferation and cell viability were measured using MTT assay and colony formation assay. Flow cytometry was adopted to test for apoptosis. Dual luciferase gene reporter assays were performed to validate the target relationship between FOXP1and miR-342 or MYCBP. The level of apoptosis-related proteins cleaved-caspase-3, Bcl-2 and Bax were measured by western blot. Compared with NHC, miR-342 expression was decreased and FOXP1 expression was up-regulated in hepatocellular carcinoma cell lines. MiR-342 could target and negatively regulate FOXP1. FOXP1 could promote the proliferation of hepatocellular carcinoma cells, positively regulate the expression of c-Caspase-3, Bax, negatively regulate Bcl-2 and inhibit apoptosis. FOXP1 can also target and positively regulate MYCBP. The expression of MYCBP was up-regulated in the hepatocellular carcinoma cell lines, while overexpression of miR-342 decreased MYCBP expression promoted by overexpression of FOXP1. MiR-342 can inhibit FOXP1/MYCBP signaling axis to regulate the members of Caspase-3 and Bcl-2 family to inhibit the proliferation and promote apoptosis of hepatocellular carcinoma cells.
探讨miR-342和FOXP1在肝癌细胞中的作用及机制。应用实时定量聚合酶链反应(QRT-PCR)检测正常肝细胞系(NHC)、肝癌细胞系(HEK-293 T)及人肝癌细胞系(HepG2、MHCC97-L、Huh7和SMMC7721)中miR-342、FOXP1和MYCBP的表达。分别在HepG2细胞中敲低或过表达miR-342和FOXP1后,采用MTT法和集落形成试验检测细胞增殖和细胞活力。采用流式细胞术检测细胞凋亡。进行双荧光素酶基因报告试验以验证FOXP1与miR-342或MYCBP之间的靶向关系。通过蛋白质免疫印迹法检测凋亡相关蛋白cleaved-caspase-3、Bcl-2和Bax的水平。与NHC相比,肝癌细胞系中miR-342表达降低,FOXP1表达上调。miR-342可靶向并负向调节FOXP1。FOXP1可促进肝癌细胞增殖,正向调节c-Caspase-3、Bax表达,负向调节Bcl-2表达并抑制细胞凋亡。FOXP1还可靶向并正向调节MYCBP。肝癌细胞系中MYCBP表达上调,而过表达miR-342可降低FOXP1过表达所促进的MYCBP表达。miR-342可抑制FOXP1/MYCBP信号轴,调节Caspase-3和Bcl-2家族成员,从而抑制肝癌细胞增殖并促进其凋亡。