蛇床子素对人前列腺癌LNCaP细胞衰老的影响及其机制研究

目的:探讨天然化合物蛇床子素对人前列腺癌LNCaP细胞衰老的影响及分子机制。方法:采用CCK8法检测蛇床子素对LNCaP细胞增殖的影响;用EdU细胞增殖检测试剂盒检测细胞增殖情况;流式细胞仪检测蛇床子素对其周期阻滞情况;β-半乳糖苷酶染色法检测蛇床子素对LNCaP细胞衰老的影响;Western blot检测衰老相关蛋白SKP2、p27的表达情况。结果:CCK8结果显示蛇床子素对LNCaP细胞的增殖有明显的抑制作用,且呈现一定剂量依赖性(P<0.05);EdU结果发现药物处理组细胞在荧光电镜下观察增殖细胞数较空白对照组逐渐减少,当蛇床子素浓度为150μmol/L时,增殖细胞数较空白对照组下降30%(P<0.01);流式细胞仪检测结果显示当蛇床子素浓度为100、150μmol/L时,阻滞于G2/M期的细胞数明显增多,并且呈现一定的浓度依赖(P<0.05);药物处理组细胞于倒置显微镜观察可见,蛇床子素浓度为150μmol/L时,β-半乳糖苷酶阳性染色细胞数较对照组明显增加(P<0.01);免疫印迹法结果显示随着蛇床子素浓度的增加,衰老相关蛋白SKP2的表达减少、p27的表达增加(P<0.05)。结论:蛇床子素可以抑制前列腺癌LNCaP细胞增殖并进一步诱导其衰老。

The molecular mechanisms of cell senescence induce by osthole in prostate cancer LNCaP cells

Objective:To investigate the effects of osthol on the senescence of human prostate cancer LNCaP cells and its molecular mechanism.Methods:The effect of osthol on the proliferation of LNCaP cells was detected by CCK8 assay.Cell proliferation was detected by EdU cell proliferation test kit.Flow cytometry detected osthol on its cycle arrest.Effect of osthol on the senescence of LNCaP cells was detected by senescence-associated beta-galactosidase（SA-β-gal）.The expression of senescence-associated proteins SKP2 and p27 was detected by Western blot.Results:CCK8 results showed that osthol inhibited the proliferation of LNCaP cells in a dose-dependent manner (P＜0.05).EdU results showed that the number of proliferating cells in the drug-treated group decreased gradually compared with the blank control group under fluorescence electron microscope.According to the statistical figure,the number of proliferating cells in the control group decreased by 30% when the concentration of osthol was 150 μmol/L (P＜0.01).The results of flow cytometry staining showed that when the concentration of osthol was 100 and 150 μmol/L,the number of cells blocked in G2/M phase increased significantly and showed a certain concentration dependence (P＜0.05).The cells in the drug treatment group were observed under an inverted microscope.With the concentration of osthol at 150 μmol/L,the number of cells in positive beta galactosylase staining increased significantly (P＜0.01).Western blot results showed that with the increase of osthol concentration,the expression of senescence related protein SKP2 decreased and the expression of p27 increased (P＜0.05).Conclusion:Osthol could inhibit the proliferation of LNCaP prostate cancer cells and further induce their senescence.