槲皮素对人胃癌细胞系SGC-7901上皮间质转化作用的初步研究

发布时间：2018-04-21 00:27

本文选题：槲皮素 + 胃癌细胞　；参考：《安徽医科大学》2017年硕士论文

【摘要】：目的:胃癌是常见的消化系统的恶性肿瘤,有很高的发生率及病死率。上皮间质转化(EMT)能够促进肿瘤细胞的侵袭和转移能力,在肿瘤的发生和进展中具有重要的作用。槲皮素(Quercetin)是一种自然界中广泛存在的黄酮类化合物,已被证实具有抗炎、抗氧化、抗肿瘤等多种生理作用。研究证实槲皮素对上皮细胞来源的恶性肿瘤具有明显的抑制作用,提示其作为抗肿瘤药物应用的潜在价值。本研究旨在探讨槲皮素对人胃癌细胞SGC-7901上皮间质转化的作用及可能机制。方法:胃癌细胞SGC-7901分为对照组(普通培养液)、LY294002组(培养液加LY294002)、槲皮素组(培养液加槲皮素);采用MTT法检测槲皮素及LY294002对胃癌细胞SGC-7901增殖的影响,依据细胞的生长抑制率(inhibition rate,IR)计算槲皮素及LY294002对胃癌细胞SGC-7901的半数抑制浓度(the median inhibitory concentration,IC50)值,参照IC50值进行后续实验;采用划痕试验检测3组胃癌细胞SGC-7901增殖迁移能力;采用Western blot检测胃癌细胞AKT、p-AKT、Snail、Vimentin及E-cadherin蛋白表达情况;采用荧光定量PCR检测3组胃癌细胞上皮间质转化相关因子Snail、Vimentin及E-cadherin m RNA的表达情况。结果:槲皮素及LY294002对胃癌细胞SGC-7901均有明显的增增殖抑制作用,槲皮素作用胃癌细胞IC50为(112.9±2.05)μmol/L,LY294002为(46.35±3.13)μmol/L;LY294002组胃癌细胞迁移距离[(0.16±0.03)mm]、槲皮素组胃癌细胞迁移距离[(0.15±0.02)mm]均低于对照组[(0.44±0.04)mm],LY294002组与槲皮素组胃癌细胞迁移距离比较差异无统计学意义;Snail、Vimentin蛋白表达量在LY294002组与槲皮素组均明显低于对照组,E-cadherin表达在LY294002组与槲皮素组高于对照组;p-AKT蛋白表达量在LY294002组与槲皮素组均明显低于对照组;AKT蛋白表达量在LY294002组、槲皮素组及对照组中无明显差异;Snail、Vimentin m RNA表达量在LY294002组、槲皮素组均明显低于对照组,E-cadherin m RNA表达量在LY294002组、槲皮素组高于对照组,差异有统计学意义);LY294002组Snail、Vimentin、E-cadherin m RNA及蛋白表达量、p-AKT蛋白表达量与槲皮素组比较差异均无统计学意义。结论:槲皮素对人胃癌细胞系SGC-7901的上皮间质转化具有抑制作用,其作用机制可能是通过对AKT信号通路的抑制实现的。
[Abstract]:Objective: gastric cancer is a common malignant tumor of digestive system with high incidence and mortality. Epithelial mesenchymal transformation (EMTT) can promote the invasion and metastasis of tumor cells and play an important role in tumorigenesis and progression. Quercetin (Quercetin) is a kind of flavonoids widely existing in nature, which has been proved to have many physiological effects such as anti-inflammatory, anti-oxidation, anti-tumor and so on. It is confirmed that quercetin has a significant inhibitory effect on malignant tumors derived from epithelial cells, indicating the potential value of quercetin as an antineoplastic drug. The aim of this study was to investigate the effect of quercetin on epithelial stromal transformation of human gastric cancer cell line SGC-7901 and its possible mechanism. Methods: gastric cancer SGC-7901 cells were divided into two groups: normal culture medium LY294002 group (culture medium + LY294002), quercetin group (culture medium + quercetin), MTT assay to detect the effect of quercetin and LY294002 on the proliferation of gastric cancer cell line SGC-7901. According to cell growth inhibition rate (IRR), the median inhibitory concentration of IC50 was calculated by quercetin and LY294002 on gastric cancer cell line SGC-7901, and the SGC-7901 proliferation and migration ability of three groups of gastric cancer cells was detected by scratch test. Western blot was used to detect the expression of Vimentin and E-cadherin protein in gastric cancer cells, and the expression of Snail-Vimentin and E-cadherin m RNA was detected by fluorescence quantitative PCR. Results: quercetin and LY294002 could significantly inhibit the proliferation of gastric cancer cell line SGC-7901. The migration distance of gastric cancer cells in quercetin treated with IC50 112.9 卤2.05 渭 mol / L LY294002 was 46.35 卤3.13 渭 mol / L LY294002 [0.16 卤0.03)mm], while the migration distance [0.15 卤0.02)mm] of quercetin group was lower than that of control group [0.44 卤0.04)mm] LY294002 group and quercetin group. There was no significant difference in migration distance between quercetin group and quercetin group. The expression of E-cadherin in LY294002 group and quercetin group was significantly lower than that in LY294002 group and quercetin group. The expression of p-AKT protein in LY294002 group and quercetin group was significantly lower than that in LY294002 group. There was no significant difference in the expression of Vimentin m RNA between the quercetin group and the control group. The expression of E-cadherin m RNA in the quercetin group was significantly lower than that in the LY294002 group, and the expression of Vimentin m RNA in the quercetin group was higher than that in the control group. There was no significant difference in the expression of E-cadherin RNA and protein between LY294002 group and quercetin group. Conclusion: quercetin can inhibit the epithelial interstitial transformation of human gastric cancer cell line SGC-7901, and its mechanism may be through the inhibition of AKT signaling pathway.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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