Effects Of Hepatocyte Growth Factor Activator Inhibitor-1 Gene On Autophagy Of Prostate Cancer PC-3 Cells

Objective This study constructed a recombinant plasmid using hepatocyte growth factor activator inhibitor-1(HAI-1)gene as a target gene,and transfected prostate cancer PC-3 cells.By detecting a series of autophagy related indicators,the effects of HAI-1 gene on the autophagy function of prostate cancer PC-3 cells were observed,and the possible mechanism of its occurrence is discussed,which provides a new theoretical basis for the occurrence and development of prostate cancer.Methods Prostate cancer PC-3 cells were transfected with recombinant plasmid pcDNA3.1(+)-HAI-1,and we also transfected plasmid pcDNA3.1(+)empty vector into PC-3 cells as an empty vector group.No plasmid transfection group was a negative control group.Total RNA and protein were extracted 72 hours after transfection,and the expressions of HAI-1 and autophagy marker Beclin 1 were detected by real-time quantitative RT-PCR.The ratio of LC3-II/I protein and the expression level of Beclin 1 protein were detected by Western blot.The ultrastructure and autophagy levels of the cells were observed by transmission electron microscopy and cell immunofluorescence.Results The results of RT-PCR showed that there is no difference in mRNA expression of HAI-1 and Beclin 1 in prostate cancer PC-3 cells between the empty vector group and the negative control group,indicating that transfection of empty plasmids have no effect on cells.By comparing the negative control group and the experimental group,the expression level of the autophagy-related gene Beclin 1 was significantly increased after transfection of the recombinant plasmid pcDNA3.1(+)-HAI-1;the results of Western blot showed that the ratio of LC3-II/I protein in the plasmid pcDNA3.1(+)-HAI-1 was significantly increased,comparing the empty vector group and the negative control group,and the expression level of Beclin 1 protein was significantly raised(P<0.05).Transmission electron microscopy showed that there is a large number of autophagosomes and autophagosomes in the cytoplasm of prostate cancer PC-3 cells in the experimental group,and fluorescently labeled autophagosomes and autophagosomes were observed by cellular immunofluorescence.Conclusion HAI-1 gene can induce autophagy in prostate cancer PC-3 cells,and the intervention of prostate cancer autophagy activity may be a potential way to treat tumors.