MicroRNA MTCO3P38 downregulates oncogene Ly6/Plaur domain-containing 1 and inhibits prolif-eration and migration of liver cancer cells
Objective To analyze the expression of Ly6/Plaur domain containing protein 1(LYPD1)in liver cancer tissues,and explore the feasibility of LYPD1 as a predictive factor for liver cancer and microRNA(miRNA)-MTCO3P38-LYPD1 pathway as a therapeutic target for liver cancer.Methods The differential expression of LYPD1 in 419 liver tissues downloaded from the cancer genome atlas(TCGA)database(including 369 cases of liver cancer tissues,50 cases of normal liver tissues,and 50 cases of paired liver cancer tissues)was analysed,and GraphPad Prism 8 software was used to calculate the area under the ROC curve using LYPD1 expression level as the predictive model for liver cancer diagnosis.Control group and miRNA MTCO3P38 group were set up,and control miRNA and miRNA MTCO3P38 were transfected into Huh7 and HCCLM9 liver cancer cells using liposomes,respectively.After 24-48 h of trans-fection,fluorescence quantitative polymerase chain reaction(qPCR)was used to detect LYPD1 mRNA expression.Western blotting was used to detect the expression level of LYPD1 protein.The cell counting kit-8(CCK-8)assay was used to analyze the proliferation ability of two groups of cells.The scratch experi-ment was used to analyze the migration ability of two groups of cells.The comparison of inter group meas-urement data was conducted using paired sample t-test or non paired sample.Results The expression value of LYPD1 was significantly higher in 369 cases of liver cancer tissues than in 50 cases of normal liver tissues(1.233±0.065 vs.0.148±0.033,t=6.082,P＜0.01).The expression value of LYPD1 was significantly higher in 50 cases of paired liver cancer tissues than in 50 cases of normal liver tissues(1.434±0.222 vs.0.148±0.033,t=5.734,P＜0.01).Using the expression values of LYPD1 in liver cancer tissue and normal liver tissue as models to predict liver cancer diagnosis,the area under the ROC curve was 0.871±0.022[95％confidence interval(CI)=0.828-0.914,P＜0.01].The qPCR results showed that the expression value of LYPD1 mRNA in the miRNA MTCO3P38 group was significantly lower than that in the control group(0.263±0.018 vs.1.033±0.088,t=7.375,P＜0.05).The Western blotting results showed that the expression of LYPD1 protein in the miRNA MTCO3P38 group was signifi-cantly lower than that in the control group(0.800±0.029 vs.2.167±0.088,t=13.480,P＜0.01).The CCK-8 experimental results showed that the expression of LYPD1 protein in the miRNA MTCO3P38 group was lower than that in the control group.The cell viability in the miRNA MTCO3P38 group was signifi-cantly lower than that in the control group(1.530±0.135 vs.3.553±0.128,t=7.695,P＜0.05).The scratch test results showed that the cell migration rate in the miRNA MTCO3P38 group was significantly lower than that in the control group(5.400±0.057 vs.9.167±0.375,t=11.850,P＜0.01).Conclusion The downregulation of the expression of LYPD1,a potential diagnostic predictor for liver cancer,by miRNA MTCO3P38 inhibits the cell proliferation and migration behavior of liver cancer cell lines.
NETL
NSTL
万方数据
