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目的探讨山楂原花青素诱导食管癌Eca109细胞凋亡的分子机制。方法常规培养食管癌Eca109细胞系并分为二甲基亚砜(DMSO)对照组和不同浓度的山楂原花青素实验组;细胞活性(cell counting kit-8,CCK-8)法检测山楂原花青素对Eca109细胞的抑制率;荧光共振能量转移(FRET)法观察山楂原花青素与细胞表面受体形成的异源二聚体的数量;Western blot法分别检测ERK5磷酸化程度和Bax、caspase-3蛋白表达水平;Annexin V-FITC/PI法检测食管癌细胞凋亡率。结果不同浓度的山楂原花青素培养Eca109细胞72h时,IC50约为275μg/ml,因此选用275μg/ml作为实验剂量;山楂原花青素与细胞表面受体形成的异源二聚体的数量呈时间依赖性,24h数量明显增多(P<0.05);ERK5磷酸化程度随着时间的延长逐渐增强(P<0.05),于36h和48h活性明显增强(P<0.01);Eca109细胞的凋亡率随着时间的推移越来越高,72 h已达48.1%(P<0.05);Bax和caspase-3蛋白表达在山楂原花青素的作用下,于48h表达最强(P<0.01)。结论山楂原花青素可促进食管癌Eca109细胞的凋亡进程。
Objective To investigate the molecular mechanism of hawthorn procyanidins inducing apoptosis of esophageal cancer Eca109 cells. Methods The esophageal cancer Eca109 cell line was routinely cultured and divided into dimethylsulfoxide (DMSO) control group and different concentrations of hawthorn proanthocyanidin experimental group. Cell counting kit-8 (CCK-8) was used to detect the effect of hawthorn proanthocyanidin on Eca109 cells . The number of heterodimers formed by the interaction between hawthorn proanthocyanidin and cell surface receptors was observed by fluorescence resonance energy transfer (FRET) method. The phosphorylation of ERK5 and the protein expression of Bax and caspase-3 were detected by Western blot. Annexin V-FITC / PI assay for esophageal cancer cell apoptosis rate. Results Different concentrations of hawthorn proanthocyanidins cultured Eca109 cells at 72h, the IC50 of about 275μg / ml, so choose 275μg / ml as the experimental dose; hawthorn proanthocyanidins and cell surface receptors formed heterodimers in a time-dependent manner, 24h (P <0.05). The phosphorylation of ERK5 gradually increased with time (P <0.05), and increased at 36h and 48h (P <0.01). The apoptosis rate of Eca109 cells increased with the passage of time (P <0.05). The expression of Bax and caspase-3 protein was the strongest at 48h (P <0.01) under the action of hawthorn procyanidins. Conclusion Hawthorn proanthocyanidins can promote the apoptosis of esophageal cancer Eca109 cells.