Mechanism Of Hydrogenated Carbazole Derivatives Inhibiting Proliferation,Migration And Invasion Of Clear Cell Renal Carcinoma Cells

Objective: To investigate the effect of hydrogenated carbazole derivative(HCZ1927)on the proliferation,migration and invasive function of renal clear cell carcinoma cells.Methods: First of all,CCK-8 was used to screen drugs that could inhibit the survival of renal clear cell carcinoma cells.The binding proteins and involved signaling pathways of HCZ1927 were predicted by bioinformatics and network pharmacology.The interaction between HCZ1927 and AKT was determined by streptomycin-agarose affinity experiments.The expressions of PI3 K,AKT,and HIF-1α in the cells were then detected by Western Blot assay.Finally,the effects of HCZ1927 on the proliferation,migration and invasion of renal clear cell carcinoma cells were studied through functional experiments.Results: We screened the efficacy of dozens of artificially synthesized naturally occurring small molecule drugs and found that the hydrogenated carbazole derivative(HCZ1927)was effective in inhibiting the survival of renal clear cell cancer cells.Structural bioinformatics combined with network pharmacology revealed that HCZ1927 could bind to AKT protein and inhibit the activity of PI3K/AKT/HIF-1αsignaling pathway in clear cell renal cell carcinoma.Western Blot results showed that after treatment with HCZ1927,the expression levels of p-PI3 K,p-AKT,AKT,and HIF-1α proteins in renal clear cell carcinoma cells were significantly decreased,but the overexpression of AKT protein could reverse the above experimental results.Experiments with CCK-8 and EDU showed that after treatment with HCZ1927,the proliferation of renal clear cell carcinoma cells was significantly inhibited.Further,through wound-healing test and Transwell test,we found that HCZ1927 could significantly weaken the migration and invasion ability of renal clear cell carcinoma cells.In addition,the overexpression of AKT protein during the treatment with HCZ1927 could effectively restore the proliferation,migration and invasion abilities of renal clear cell carcinoma cells.In summary,we believe that the hydrogenated carbazole derivative(HCZ1927)most likely inhibited the proliferation,migration,and invasion of renal clear cell carcinoma cells by inhibiting the PI3K/AKT/HIF-1αsignaling pathway.Hydrogenated carbazole derivative(HCZ1927)was able to inhibit the survival of renal clear cell carcinoma cells.In vitro studies have revealed that HCZ1927 inhibits the proliferation,migration,and invasion of renal clear cell carcinoma cells as well as epithelial-mesenchymal transition(EMT).In renal clear cell carcinoma cells,we observed that HCZ1927 negatively regulated the PI3K/AKT/HIF-1α pathway and that it combined with AKT to inhibit AKT protein expression.Conclusion: In this study,bioinformatics and network pharmacology analysis were combined with in vitro experiments,and we found for the first time that HCZ1927 could down-regulate the activity of PI3K/AKT/HIF-1α signaling pathway to inhibit the proliferation,migration and invasion of renal clear cell carcinoma cells.Therefore,we believe that HCZ1927 is involved in a variety of potential molecular mechanisms that provide a new approach to studying the potential drug development of clear cell renal cell carcinoma.