龙葵碱调控p-AKT、Cleaved caspase-3和p53蛋白表达抑制卵巢癌细胞增殖实验研究

目的:研究龙葵碱调控磷酸化蛋白激酶B((Phosphorylated protein kinase B,p-AKT)、Cleaved caspase-3和p53蛋白表达抑制卵巢癌细胞增殖诱导其凋亡。方法:将SKOV3卵巢癌细胞分为模型对照组、龙葵碱15μmol/L、龙葵碱10μmol/L、龙葵碱5μmol/L组,同时选择正常卵巢细胞作为正常对照组,并进行培养。使用倒置显微镜观察卵巢癌细胞的形态,使用MTT法、流式细胞仪、蛋白质印迹法(Western blot)检测不同浓度龙葵碱对卵巢癌细胞的增殖率、细胞凋亡及Cleaved caspase-3和p53蛋白表达。结果:倒置显微镜观察SKOV3卵巢癌细胞形态,随着龙葵碱的浓度越高,细胞凋亡的形态学表现越明显。正常对照组的细胞凋亡率和细胞增值率显著低于其他各组,具有统计学差异(P<0.05)。随着时间的延长和药物浓度的提高SKOV3卵巢癌细胞的生长抑制作用越来越显著,与模型对照组相比,具有统计学意义(P<0.05)。模型对照组p-AKT、Cleaved caspase-3的表达显著低于龙葵碱5μmol/L、10μmol/L、15μmol/L组,正常对照组低于模型对照组,龙葵组p53蛋白显著高于模型对照组,正常对照组p53蛋白低于卵巢癌组,有统计学意义(P<0.05)。龙葵碱5μmol/Lp-AKT、Cleaved caspase-3的表达显著低于龙葵碱10μmol/L、15μmol/L组,p53蛋白表达高于其他两组浓度,随着龙葵碱浓度的升高,p-AKT、Cleaved caspase-3表达逐渐升高,p53蛋白表达逐渐降低,有统计学差异(P<0.05)。结论:龙葵碱可能通过调控p-AKT、Cleaved caspase-3和p53蛋白的表达而抑制SKOV3卵巢癌的增殖能力,诱导其凋亡,其能力呈现浓度依赖。

Solanum nigrum inhibits proliferation and promote apoptosis of ovarian cancer cells through regulates the expression of p-AKT,Cleaved caspase-3 and p53 protein

Objective:To investigate solanum nigrum inhibits proliferation and promote apoptosis of ovarian cancer cells through regulates the expression of Phosphorylated protein kinase B(p-AKT),Cleaved caspase-3 and p53 protein.Methods:SKOV3 ovarian cancer cells were divided into model control group,5μmol/L group,10μmol/L group,and 15μmol/L group,simultaneous selection of normal ovarian cells as normal control group.The cells were cultivated in 4 groups.The morphology of SKOV3 ovarian cancer cells was observed by inverted microscope.Detection of cell proliferation by MTT colorimetry.Detection of apoptosis efficiency by flow cytometry.Detection of p-AKT,Cleaved caspase-3 and p53 protein expression by western blotting.Results:The higher the concentration of solanum alkaloids,the more obvious morphological manifestations of apoptosis under the inverted microscope.The apoptosis rate and cell proliferation rate of the normal control group were significantly lower than those of the other 4 groups(all P<0.05).With the prolongation of time and the increase of drug concentration,the growth inhibition of SKOV3 ovarian cancer cells became more and more significant,and the difference was statistically significant compared with the model control group(P<0.05).The expression of p-AKT and Cleaved caspase-3 of the model control group were significantly lower than 5μmol/L group,10μmol/L group,15μmol/L group(all P<0.05),and the normal control group were lower than the model control group(P<0.05).The p53 protein of the 5μmol/L group,10μmol/L group,15μmol/L group was significantly higher than that of the model control group(all P<0.05),and the p53 protein of the normal control group was lower than that of the other 4 group(all P<0.05).The expression of p-AKT and Cleaved caspase-3 of the 5μmol/L group were significantly lower than those in the 10μmol/L group and 15μmol/L group,and the expression of p53 protein was higher than that of other 2 groups.with the concentration of solanum alkaloids increased,the caspase-3 expression of p-AKT and Cleaved caspase-3 increased gradually,and the expression of p53 protein decreased gradually,significantly(P<0.05).Conclusion:Solanum nigrum inhibits proliferation and promote apoptosis of ovarian cancer cells through regulates the expression of p-AKT,Cleaved caspase-3 and p53 protein in a concentration-dependent manner.