Isovitexin Inhibit The Tumorigenicity By Regulate Expression Of MnSOD/p-CaMK Ⅱ In NCI-H460 Cells

Objetive:To study the isovitexin(ISOV)on MnSOD/p-CaMK Ⅱ signal pathway,and tumorigenicity in human non-small cell lung cancer H460 cell derived stem-like cells(LCSLC).Methods:1.Non-small cell lung cancer H460 cells were cultured using DMEM medium containing 10%serum in vitro,and LCSLC were obtained by ultra-low adherence culture with free.senum stem cell conditioned medium.LCSLC were dealed with different concentrations of ISOV(l,3,10 μg/mL).The protein expression levels of MnSOD,(p-CaMK Ⅱ)and cancer stem cell markers CD 133,CD44,ALDH1 were analyzed by western blot.The tumorigenicities of LCSLC were detected by tumor sphere formation assay,wound healing assay,soft agar colony forming assay and tanswell invasion assay.The glycolysis metabolism kit was used to determine lactate metabolism level.2.MnSOD knockdown of LCSLC was used by infection of adenovirus expressing MnSOD shRNA.The expressions of MnSOD,p-CaMKⅡ,cancer stem cell markers(CD133,CD44 and ALDH1)proteins were analysed by western blot.The tumorigenicity of LCSLC were tested via sphere formation assay,soft agar colony formation assay,invasion and migration assay.The metabolism glycolysis kit was used to determinate level of lactic acid metabolis.3.LCLSC were treated by MnSOD shRNA combined with ISOV.The protein expressions of MnSOD,P-CaMK Ⅱ,cancer stem cell markers(CD133,CD44 and ALDH1)were analyzed by western blot,The tumorigenicities of LCSLC were detected by tumor sphere formation assay,soft agar colony forming assay,wound healing assay,and transwell invasion assay.The glycolysis metabolism kit was used to determine lactate metabolism level.Results:1.Compared with H460 cells,the protein expressions of MnSOD,p-CaMK Ⅱ and cancer stem cell markers(CD133,CD44 and ALDH1),in vitro tumorigenicity and level of lactic acid metabolism were increased in LCSLC.2.ISOV inhibited the protein expressions of MnSOD,p-CaMK Ⅱ,cancer stem cell markers(CD133,CD44 and ALDH1),and in vitro tumorigenicity and level of lactic acid metabolism in LCSLC,in concentration-dependent manner.3.MnSOD shRNA reduced the protein expressions of MnSOD,p-CaMKⅡ,cancer stem cell markers(CD133,CD44 and ALDHI),in vitro tumorigenicity and level of lactic acid metabolism in LCSLC.4.MnSOD shRNA combined with ISOV cooperatively decreased the protein expressions of MnSOD,p-CaMKⅡ,cancer stem cell markers(CD133,CD44 and ALDH1),in vitro tumorigenicity and level of lactate metabolism in LCSLC.Conclusions1.ISOV inhibits in vitro tumorigenicities and aerobic glycolysis of LCSLC derived from H460 cells,in concentration-dependent manner.2.ISOV inhibits in viztr tumorigenicities and aerobic glycolysis of LCS’LC derived from H460 cells is involved in blockage of MnSOD/p-CaMK Ⅱ signaling.