蜂胶黄酮对人结肠癌细胞PDE4D及Gadd45G基因表达的影响

Effects of propolis flavonoids pinobanksin-3-acetate on expression of PDE4D and Gadd45G gene in human colon cancer cell line

目的探讨蜂胶黄酮(PB3A)对人结肠癌SW480细胞生长的抑制作用及机制。方法将培养的人结肠癌SW480细胞分为PB3A组及对照组。PB3A组予100μg/mL PB3A干预,对照组不干预。干预后两组均培养24 h,倒置显微镜观察细胞形态学变化;采用实时荧光定量PCR法和蛋白免疫印迹法检测细胞磷酸二酯酶4D(PDE4D)、生长阻滞和DNA损伤诱导基因G(Gadd45G)mRNA和蛋白表达。结果干预后倒置显微镜下可见PB3A组细胞缩小、皱缩、折光性减弱,细胞内出现颗粒状物质;对照组无明显变化。PB3A组PDE4D mRNA及蛋白表达水平均低于对照组,Gadd45G mRNA及蛋白水平表达均高于对照组(P均<0.01)。结论 PB3A能抑制SW480细胞生长,诱导其凋亡;其作用机制可能为下调PDE4D mRNA和蛋白表达,上调Gadd45G mRNA和蛋白表达。

Objective To investigate the inhibitory effect of propolis flavonoids pinobanksin-3-acetate （PB3A） on the human colon cancer cell line SW 480 and to explore the underlying mechanism .Methods The cultivated human colon cancer cell line SW480 was randomly divided into the PB3A group and the control group .Cells in the PB3A group were in-tervened by 100 μg/mL PB3A, and the control group without intervention .We observed the cellular morphology by invert microscope after incubation for 24 h of the two groups .Real-time fluorescence quantitative polymerase chain reaction and Western blotting were performed to measure the expression of phosphodiesterase 4D （PDE4D） and growth arrest and DNA damage-inducible gene G （Gadd 45G） at mRNA and protein levels.Results After treated with 100 μg/mL PB3A, the colon cancer cells became shrunk , smaller, refraction decreased with granular material appeared intra-celluler in the PB3A group.In comparison with those in the control group , the mRNA and protein expression of PDE4D was down-regulated, while the mRNA and protein expression of Gadd 45G was up-regulated （all P＆lt;0.01）.Conclusions PB3A inhibited the proliferation of SW480 cells and induced the apoptosis .Its possible mechanisms may be related to down-regulation of PDE4D expression and up-regulation of Gadd45G expression.