Effect Of Dihydroartemisinin On UHRF1 Of Prostate Cancer Xenograft Model In Nude Mice And Its Regulatory Mechanism

Objective:This study aims to investigate the effect of dihydroartemisinin on Ubiquitin-like with PHD and ring finger domains 1（UHRF1）of prostate cancer PC-3 cells xenograft model in nude mice,and to further explore its regulatory mechanism of methylation.Methods:Prostate cance PC-3 cells were transplanted to establish the classic xenotransplantation mode.This nude mice were randomly divided into 4 groups:the control group,the solvent group（DMSO group）and the DHA group（50umol/kg,100umol/kg）.After tumor formation,we researched on it with DHA intervention once every two days.On the 13th day,samples were collected to calculate the rate of tumor inhibition and measured in vitro.The morphological manifestations of apoptosis in cancer tissues were observed by light microscope and electron microscope;the apoptosis of tumor cells and the apoptosis rate were observed by TUNEL;expression of UHRF1,DNMT1 and p16^INK4AmRNA were detected by Quantitative real-time PCR;expression at protein levels of UHRF1,DNMT1 and p16^INK4Awere observed by Western blotting;The cellular location and expression of UHRF1,DNMT1 and p16^INK4Aprotein were detected by immunohistochemical methods;methylation levels in the p16^INK4ACpG islands was determined by bisulfite genomic sequencing.Results:1.The tumor inhibition rate suggested that DHA could inhibit the progression of prostate cancer PC-3 cells in nude mice.2.The morphological changes of apoptosis and necrosis induced by DHA in PC-3cells were observed by light microscopy and electron microscopy.3.TUNEL showed that dihydroartemisinin could induce apoptosis in PC-3 cells.4.RT-PCR,western blot and immunohistochemistry showed that DHA could decrease the expression of UHRF1 and DNMT1 in PC-3 cells and increase the expression of tumor suppressor p16^INK4A.5.BGS showed that the methylation level of the promoter region of tumor suppressor p16^INK4Adecreased significantly after DHA acted on pc-3 cells.Conclusion:DHA can effectively inhibit the progression of tumor of prostate cancer in nude mice.It may result from down-regulating the expression of UHRF1 and DNMT1,inducing the demethylation of the promoter region of tumor suppressor p16^INK4Aand restoring its expression,so as to exert its effect of anti-cancer and induce the apoptosis of PC-3 cells in prostate cancer.