miR-181a在肝癌细胞化疗耐药中的作用及其相关靶基因预测OA

Objective To investigate the role of miR-181a in chemotherapy resistance of hepatocellular carcinoma(HCC)cells and to predict its related target genes.Methods The doxorubicin-resistant human HCC cell line HepG2/ADM and the human HCC cell line HepG2 were cultured and collected for experiments.The two cell lines were divided into the non-resistant group(HepG2 cells),non-resistant+high expression group(HepG2 cells transfected with miR-181a ag-omir),resistant group(HepG2/ADM cells),resistant+low expression group(HepG2/ADM cells transfected with miR-181a antagomir),and negative control group(cells transfected with blank vector,HepG2/ADM and HepG2 cells),re-spectively.The miR-181a was detected by qPCR.The non-resistant group,non-resistant+high expression group,resistant group,and resistant+low expression group were treated with different concentrations of doxorubicin,cisplatin,and 5-flu-orouracil.The cell proliferation inhibition rate was measured by MTT assay,and the half-maximal inhibitory concentration(IC50)was calculated.After obtaining the IC50 values,HepG2/ADM cells were divided into the blank group(no drug and no transfection),low expression group(transfected with miR-181a antagomir),NC group(transfected with miR inhibitor NC),ADM group(treated with IC50 concentration of doxorubicin),and low expression+ADM group(transfected with miR-181a antagomir+treated with IC50 concentration of doxorubicin),respectively.The apoptosis was detected by flow cy-tometry.The GEO database was used to analyze the expression profile of the GSE206501 dataset,screening for differentially expressed genes between oxaliplatin-resistant and oxaliplatin-sensitive liver cancer patients.These were intersected with miR-181a target genes predicted by online databases to obtain hub target genes of miR-181a.The GSCA database was used to analyze the relationship between hub target genes and survival of HCC patients.The GEPIA2 database was used to ana-lyze the correlation between survival-related hub target genes and clinical stages.Combined with the differential expression gene screening results from the GSE206501 dataset,the target genes of miR-181a were finally determined.The GSCA data-base was used to perform pathway activity analysis on the target genes of miR-181a.Results The expression of miR-181a in the resistant group was higher than that in the non-resistant group(P＜0.01).The IC50 values of doxorubicin,cisplatin,and 5-fluorouracil in the resistant+low expression group were lower than those in the resistant group,while the IC50 values in the non-resistant+high expression group were higher than those in the non-resistant group(all P＜0.05).The apoptosis rates of HepG2/ADM cells in the low expression group,ADM group,and ADM+low expression group increased sequen-tially and were all higher than those in the blank group and NC group(all P＜0.01).The screened target genes of miR-181a included CCNB1,KIF2C,MELK,NEK2,NDUFV3,TOP2A,and TPX2.These genes activated apoptosis,cell cycle,and epithelial-mesenchymal transition pathways,and inhibited the androgen receptor,estrogen receptor,RAS/MAPK,and RTK pathways.Conclusions The miR-181a is up-regulated in the drug-resistant cells of HepG2/ADM,and plays a criti-cal role in mediating multidrug resistance in HCC chemotherapy,potentially through the regulation of apoptosis.The target genes may include CCNB1,KIF2C,MELK,NEK2,NDUFV3,TOP2A,and TPX2.