Effect of germacrone on proliferation of gastric cancer cells by regulating PRAS40/p-PRAS4 through HBXIP
Objective To study the mechanism of the regulation of gastric cancer cell proliferation by germacrone through HBXIP.Methods Gastric cancer tissues and paired paracancer tissues were collected from 20 patients undergoing gastrectomy.The expression of HBXIP,prolin-rich protein kinase B substrate protein(PRAS40)with relative molecular weight of 4×104 and p-PRAS40 were detected by immunohistochemical and Western blotting.The correlation between PRAS40 and p-PRAS40 expression and HBXIP expression in 20 cancer tissues was analyzed by correlation Analyze.After overexpressing or knocking down HBXIP in MGC803 and SGC7901,the change of cell proliferation was detected by MTT.After HBXIP was overexpressed or knocked-down in MGC803 and SGC7901,the apoptosis was detected by flow cytometry and Hoechst 33285,and the expression of HBXIP,PRAS40 and p-PRAS40 were detected by Western blotting.MGC803 and SGC7901 were treated with 0,50,100,150,200,250 μmol·L-1 germacrone for 48 h.The proliferation ability of the cells was detected by MTT and the apoptosis level was detected by flow cytometry.The expression of HBXIP,PRAS40 and p-PRAS40 in MGC803 and SGC7901 were treated with 150 μmol·L-1 germacrone for 48 h were detected by Western blotting.Results The expression of HBXIP,PRAS40 and p-PRAS40 in gastric cancer tissues was significantly higher than that in adjacent gastric cancer tissues(P＜0.05),and the expression of PRAS40 and p-PRAS40 was positively correlated with the expression level of HBXIP.After knocking down HBXIP in MGC803 and SGC7901,compared to the NC group,cell proliferation was decreased and cell apoptosis was increased(P＜0.05).The expression of PRAS40 and p-PRAS40 was also significantly decreased or increased after HBXIP was knocked down or overexpressed in MGC803 and SGC7901.Germacrone inhibited the proliferation and induced apoptosis in a dose-dependent manner(P＜0.05).Germacrone inhibited the expression of HBXIP,PRAS40 and p-PRAS40.Conclusion Germacrone can promote the proliferation of gastric cancer cells by down-regulating HBXIP,and the mechanism may be related to PRAS40 and p-PRAS40 down-regulated by HBXIP.
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