Expression Of SOX5?GATA3 In Classic Kaposi's Sarcoma And Its Effect On Proliferation And Apoptosis

Objective The aim of this study is to uncover the regulation network mianly associated with SRY-related high-mobility-group box 5(SOX5)and GATA-binding protein 3(GATA3)in the development of Kaposi's Sarcoma(KS),include cellar proliferation and apoptosis.Thus provide a possible treatment target for KS.MethodImmunohistochemistry and real-time PCR were used to detect the expression of SOX5 and GATA3 in tumors and adjacent normal tissues in classic KS patients;Further,Kaposi's sarcoma-associated herpesvirus(KSHV)was used to infecte cell lines iSLK-219 and iSLK-BAC,and KSHV was used as uninfected control toc determine the expression difference under such transfection.Further sequenced downstream effected genes associated with proliferation and apoptosis were manipulatedby vectors transfection.CCK-8 kit and flow cytometry were used to detect the effect of SOX5 and GATA3 in cellar vitality and apoptosis in infected Western blotting was used to detect the expression of proliferation-related proteins PCNA,PKC,apoptosis-related proteins Bcl-2 and C-PARP in SOX5 and GATA3 overexpressed KSHV cell line;The sequencing method was used to screen the differential genes related to cell proliferation and apoptosis after SOX5 and GATA3 overexpression,and to explore the possible pathway related with proliferation and apoptosis effected by SOX5 and GATA3 Detail mechanism of how SOX5 and GATA3 participate in the pathological development of KS is thus uncovered.Results1.Immunohistochemistry and real-time PCR revealed that SOX5 and GATA3 showed low expression in tumor tissues in classical KS patients,and showed a statistically significant difference whencompared with normal tissues adjacent to the tumor(P < 0.01),and the expression of SOX5 and GATA3 were highly related to regional difference and course of disease of KS(P < 0.05).2.The expression of SOX5 and GATA3 in KSHV-infected cell lines was lower than that of uninfected control cell lines(P < 0.01).3.The results of real-time PCR showed that most of the SOX5 and GATA3 target genes were related to proliferation and apoptosis.4.The results of CCK-8 showed that after transfecting SOX5 and GATA3 into iSLK-219 and iSLK-BAC cells respectively,the cell growth speed was decreased than that of the empty vector transfected group(P < 0.01).This suggested that SOX5 and GATA3 can inhibited the proliferation of KSHV-infected cell lines.5.When SOX5 and GATA3 were transfected into iSLK-219 and iSLK-BAC cells,respectively,the apoptotic rate was significantly decreased compared with the empty vector transfected group(P < 0.05).6.Overexpressing SOX5 and GATA3 resulted in decrease expression of C-PARP,anti-apoptotic protein Bcl-2and proliferation-related proteins PCNA and PKC.7.Transcriptome sequencing analysis showed that SOX5 and GATA3 are highly associated with JAK-STAT signaling pathway.8.Overexpression of GATA3 in iSLK-219 cells did not change total protein level of ERK and STAT3 compared with cells transfected with empty vector;whereas decrease p-ERK and p-STAT3 significantly.Conclusion1.SOX5 and GATA3 were lowly expressed in both classical KS tissues and KSHV infected cells.2.SOX5 and GATA3 can inhibit KSHV-infected cell lines proliferation and promote cell apoptosis.3.GATA3 can inhibite cell proliferation and promote apoptosis through inhibits phosphorylation of ERK and STAT3 in KSHV-infected cells.