The Roles And Mechanisms Of SIVA In Inhibiting The Proliferation And Inducing Apoptosis Of Hepatocellular Carcinoma Cells By Plumbagin

Purpose: Plumbagin(PL)is thought to be a bioactive phytochemical drug and exerts antitumor effect on various cancers.However,few studies focus on the antitumor activity of PL on hepatocellular carcinoma(HCC).In this study,we firstly investigated the antitumor activity of PL on HCC cells and further investigated the molecular mechanism of its antitumor activity against HCC both in vitro and in vivo.Finally,we assessed the relationship between the expression of SIVA and HCC progression.Methods: We evaluated the antiproliferative activity of PL through CCK-8,Ed U and colony forming test.Then we analyzed the cell cycle and apoptosis by flow cytometer.Western blot was used to detect the expression of apoptosis related protein,SIVA and m TOR pathway.Wound healing and Transwell were applied to test the cell migration and invasion.We also performed RNA-seq to determine the gene expression profiles and overexpressed or knocked down SIVA to validate its role in PL's antitumor activity.Regarding animal experiment,we built xenograft model in BALB/c nude mice using LM3 cells.Then immunohistochemistry was performed to study the antitumor activity and the expression of SIVA and m TOR in PL-treated group.The cases of TMA were divided into “Low” and “High” groups according to the results of immunohistochemistry and various analyzed were performed between these two groups.We facilitated the online tools to further investigated the relationship between the expression of SIVA and HCC progression.Results: PL can exert antitumor power on inhibiting proliferation,migration,invasion and inducing apoptosis of liver cancer cells in vitro.Further experiment demonstrated that PL could inhibit the expression of SIVA and subsequently downregulate m TOR signaling pathway.Upregulating the expression of SIVA will alleviate the antitumor activity of PL on liver cancer,which confirmed the important role of SIVA/m TOR signaling pathway in the antitumor activity of PL.In vivo immunohistochemistry also demonstrated that PL could inhibit SIVA/m TOR signaling pathway in tumor tissues.Analyses of TMA and datasets from GEPIA and KM-plotter revealed that the population with lower expression of SIVA had longer overall survival.Conclusion: Our promising results showed that PL could inhibit proliferation and induce apoptosis of hepatic cancer through inhibiting SIVA/m TOR signaling pathway for the first time,which indicated that PL might be a good candidate against liver cancer,especially for patients who have higher expression of SIVA.