Up-regulation Death Receptor 4 Enhance TRAIL-induced Apoptosis In Human Renal Cancer Cells

Background:Renal cell carcinoma(RCC)has a high incidence,which seriously threatens human life and health.For patients with advanced and metastatic kidney cancer,safe and effective treatment methods still need further research.Tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)can highly selectively induce apoptosis of cancer cells and has little toxicity to normal tissues,so it can be used as a safe anti-cancer drug.However,in more and more studies,it has been found that cancer cells have obvious resistance to apoptosis induced by TRAIL.Death receptor is the target of TRAIL to induce apoptosis of cancer cells.The protein expression level of death receptor is considered to be positively related to the sensitivity of TRAIL.Increasing the expression level of death receptors in kidney cancer cells can restore the sensitivity of kidney cancer cells to TRAIL.Objectives:(1)It is proved that up-regulating the protein expression level of death receptor can restore the sensitivity of renal cell carcinoma to TRAIL.(2)Looking for safe and effective TRAIL sensitizers to provide the possibility for the clinical application of TRAIL.(3)In-depth exploration of the mechanism of andrographolide and PFT-? combined with TRAIL to inhibit the proliferation and migration of renal cancer cells and induce programmed apoptosis of renal cancer cells.(4)Explore the molecular mechanism of renal cancer cell resistance to TRAIL,and look for treatment strategies.Methods:Through the mRNA expression data of DR4 and DR5 in the database,analyze the expression differences of death receptors in renal cancer cells and normal kidney tissues;detect tumor cell viability by MTS method;detect cell proliferation experiments,Edu staining proliferation experiments and cloning experiments The proliferation ability of tumor cells;the migration ability of tumor cells was detected by the plate scratch healing test.Detect cell cycle and apoptosis by flow cytometry;detect the expression level of various related proteins by western blot;prepare target gene-silencing cell lines by small-molecule RNA interference and lentiviral packaging infection methods;use fluorescence real-time quantitative PCR technology Detect changes in cell transcription levels.Results:(1)The expression level of death receptor protein in kidney cancer is higher than that in normal tissues,so TRAIL can be used as a potential therapeutic agent to target kidney cancer.The protein expression level of DR5 in renal cancer cells was significantly higher than that of normal tissues,but the protein expression level of DR4 was not significantly higher than that of DR5.(2)The increased expression level of DR4 protein can enhance the sensitivity of renal cell carcinoma to apoptosis induced by TRAIL.(3)The combined application of andrographolide or PFT-? and TRAIL can significantly inhibit the cell viability,proliferation and migration ability of renal cancer cells,induce renal cancer cell cycle arrest and cell senescence,and induce Caspase-dependent programmed apoptosis of renal cancer cells.Die.(4)Knockdown mutant p53 induces an increase in the expression level of DR4 protein in renal cancer cells and restores the sensitivity of renal cancer to TRAIL.Conclusion:The sensitivity of kidney cancer to TRAIL can be restored by up-regulating DR4.Andrographolide and PFT-? are safe and effective potential TRAIL sensitizers,and their combined application becomes a potential kidney cancer treatment drug.Mutant p53 induces a decrease in the expression of DR4 in renal cancer cells,which is one of the mechanisms of renal cancer resistance to TRAIL.