金水鲜联合放疗对A549细胞血管生成及凋亡相关蛋白表达关系的研究

摘要

目的：研究金水鲜联合放疗对肺腺癌A549细胞实验处理后的细胞存活力,以及抗血管生成蛋白表达与凋亡相关蛋白表达的关系变化。方法将生长状态良好的A549细胞按处理因素的不同分为4组,生理盐水(NC)组、金水鲜(JSX)组、放射治疗垣金水鲜(RT+JSX)组、放射治疗(RT)组,用CCK8试剂盒处理4 h后酶标仪测OD450值,收集细胞裂解后测定蛋白浓度并蛋白凝胶(WB)电泳,分析血管促进因子蛋白血管生成因子(VEGF)/胎盘源性生长因子(PIGF)和突变型P53以及促凋亡蛋白FAS蛋白的表达变化。结果淤CCK8检测各组经过处理后细胞的增殖活力,RT+JSX组细胞的增殖活力最低,与JSX组比较,差异有高度统计学意义(P<0.01)；JSX组与NC组比较,差异有统计学意义(P<0.05)。于Western-blot结果显示在第48小时,JSX组VEGF蛋白的表达下降明显；而在RT+JSX组中促凋亡蛋白FAS的表达明显增加,突变型P53蛋白的表达下降,与JSX组比较,差异有高度统计学意义(P<0.01),提示联合使用金水鲜增强了放疗自身的促凋亡作用。结论金水鲜联合放疗可能发挥药物单独具有的血管生成抑制作用,并能增强单用放疗的细胞促凋亡效果,考虑为金水鲜在影响血管生成方面能增加放疗的敏感性,激活细胞凋亡相关因子FAS和下调突变型P53,具体作用机制有待进一步证实。%Objective To study cell viability, anti angiogenic protein expression and apoptosis related protein expres-sion after the experimental treatment of Jinshuixian combined with radiotherapy on lung A549 cells. Methods A549 cells in good condition were divided into 4 groups including physiological saline (NC) group, Jinshuixian (JSX)group, radiotherapy + Jinshuixian (RT+JSX) group, radiation therapy (RT) group, according to the treatment factors according to processing factors, OD450 were measured after CCK8 kit for 4 h. Determination of protein concentration and WB electrophoresis were collected after cell lysis. VEGF/PIGF and mutant P53 protein and the expression of pro-apoptotic protein FAS were measured after the process. Results [1] The cell proliferation activity of each group after Goldwater fresh and radiotherapy, cell proliferation activity of RT+JSX group was the lowest, compared with that of JSX group, the differences were statistically significant (P< 0.01); cell proliferation activity of JSX group was compared with that of NC group, the difference was statistically significant (P< 0.05). [2] Western-blot experiments showed that in the first 48 hours, the expression of VEGF protein was decreased significantly in JSX group; the expression of pro-apoptotic protein FAS in RT+JSX group was increased, the expression of mutant P53 protein was decreased, compared with JSX group, the differences were statistically significant (P< 0.01), it prompted Jinshuixian enhanced the pro-apoptotic ef-fects of radiotherapy. Conclusion Jinshuixian combined with radiotherapy may has stronger inhibitory effect on angio-genesis, and can enhance the single cell apoptosis effect of radiotherapy, which is considered as the sensitivity of ra-diotherapy. This combined therapy can also activate the apoptosis related factor FAS and adjustable mutant P53. But the specific mechanism needs further confirmation.