杨梅素对人卵巢癌SKOV3细胞的凋亡诱导作用

目的:观察杨梅素对人卵巢癌SKOV3细胞的生长抑制及凋亡诱导作用,并探讨其可能机制。方法:体外培养人卵巢癌SKOV3细胞,按杨梅素给药浓度将细胞分为对照组和低、中、高浓度杨梅素组,杨梅素给药浓度分别为0、20、40和60 mg·L^-1。MTT法检测各组SKOV3细胞存活率;激光共聚焦显微镜观察各组SKOV3细胞核形态变化并计算异常细胞核比率;免疫荧光法检测各组SKOV3细胞中凋亡相关蛋白-活化半胱氨酸蛋白酶3(active Caspase-3)的表达;Western blotting法检测各组SKOV3细胞中DNA损伤相关蛋白γ-H₂AX的表达水平。结果:与对照组比较,低、中和高浓度杨梅素组SKOV3细胞存活率明显下降(t=-12.35,t=-11.42,t=-10.21,P<0.05);激光共聚焦显微镜下各浓度杨梅素组细胞出现核皱缩等凋亡形态学改变,且细胞中异常细胞核比率高于对照组(t=6.87,t=9.35,t=7.22,P<0.05),高浓度杨梅素组异常细胞核比率高于低、中浓度杨梅素组(t=9.55,t=7.25,P<0.05);随着杨梅素给药浓度的增加,SKOV3细胞中active Caspase-3的表达水平有升高趋势,各浓度杨梅素组SKOV3细胞中γ-H₂AX的表达水平高于对照组(t=5.41,t=3.21,t=6.58,P<0.05)。结论:杨梅素可抑制人卵巢癌SKOV3细胞增殖并诱导其凋亡,DNA双链断裂可能是其促凋亡的机制之一。

Induction effect of myricetin on apoptosis of human ovarian cancer SKOV3 cells

Objective To observe the induction effect of myricetin on the apoptosis and the inhibitory effect on the growth of the human ovarian cancer SKOV3 cells,and to explore the probable mechanisms. Methods The human ovarian cancer SKOV3 cells were cultured in vitro.The cells were divided into control group,low,middle and high concentrations of myricetin groups,which were treated with different concentrations (0,20,40 and 60 mg·L^-1) of myricetin.Then the survival rates of cells were tested by MTT method;Laser scanning confocal microscope was used to observe the morphological changes,and the rates of abnormal necleuses were calculated.The expressions of active Caspase-3 in the SKOV3 cells in various groups were observed by immunofluorescence method.The expression levels of γ-H₂AX in the SKOV3 cells in various groups were detected by Western blotting method. Results Compared with control group,the survival rates of the SKOV3 cells in myricetin groups were significantly decreased(t=-12.35,t=-11.42,t=-10.21,P<0.05).The morphology of nucleuses of SKOV3 cells in various groups had some changes for the cause of apoptosis which were observed by laser scanning confocal microscope,nuclear shrinkage for example.The rates of abnormal nucleuses in low,middle and high concentration of myricetin groups were obviously higher than that in control group(t=6.87,t=9.35,t=7.22,P<0.05),and the rate of abnormal nucleus in high concentration of myricetin group was higher than those in low and middle concentrations of myricetin groups(t=9.55,t=7.25,P<0.05).The expression levels of active Caspase-3 in the SKOV3 cells in myricetin groups had increasing tendency,and the expression levels of γ-H₂AX in the SKOV3 cells in myricetin groups were higher than that in control group(t=5.41,t=3.21,t=6.58,P<0.05). Conclusion Myricetin can inhibit the proliferation of SKOV3 cells and induce the apoptosis;its possible mechanism may be related to DNA double strands break.