青藤碱对人胃癌SGC-7901细胞EMT的作用机制研究

发布时间：2018-04-09 22:35

本文选题：青藤碱　切入点：人胃癌细胞SGC-7901　出处：《河北医科大学》2015年硕士论文

【摘要】：目的:研究青藤碱(SIN)对人胃癌细胞SGC-7901增殖、侵袭以及转移相关基因和蛋白表达的影响。方法:1细胞存活率检测:用不同浓度SIN处理人胃癌SGC-7901细胞后,用WST-1法检测不同处理时间的细胞存活率。2 MMP表达:用不同浓度SIN处理人胃癌SGC-7901细胞,ELISA检测MMP-2和MMP-9在细胞培养基中的含量变化。3 RT-PCR检测不同浓度SIN对人胃癌SGC-7901细胞相关EMT基因mRNA表达水平的影响。4 Western blotting检测不同浓度SIN对人胃癌SGC-7901细胞相EMT蛋白表达水平的影响。5 Western blotting检测不同浓度SIN对人胃癌SGC-7901细胞核内NFκB p65蛋白表达水平的影响。结果:1采用不同浓度的SIN处理人胃癌SGC-7901细胞,检测结果表明:处理24 h后,1 mmol/L、2 mmol/L、2.5 mmol/L和3mmol/L处理组的细胞存活率显著低于对照组(P0.05)。处理48 h后,0.5 mmol/L、1 mmol/L、2 mmol/L、2.5 mmol/L以及3 mmol/L处理组的细胞存活率显著低于对照组(P0.05),且对青藤碱具有时间和剂量依赖性。2 SIN处理后,可降低人胃癌SGC-7901细胞培养基MMPs的含量。各SIN处理组培养基中MMP-2含量显著低于对照组(P0.05),1 mmol/L和2 mmol/L处理组的MMP-9含量降低(P0.05)。3 RT-PCR和Western Blotting结果表明:随SIN浓度升高,各处理组人胃癌SGC-7901细胞中MMP-2 m RNA、蛋白表达以及MMP-9 mRNA表达丰度均显著降低(P0.05)。其中0.5mmol/L、1 mmol/L和2 mmol/L三个处理组MMP-9蛋白表达也显著降低(P0.05);与对照组相比,0.5mmol/L、1mmol/L和2 mmol/L处理组Snail基因和蛋白表达显著降低(P0.05);与对照组相比,0.5 mmol/L、1mmol/L和2 mmol/L处理组E-cadherin基因和蛋白表达显著升高(P0.05)。4 Western Blotting结果发现,随着SIN浓度的升高,SGC-7901细胞核中NFκB p65蛋白表达呈现降低趋势。与对照组相比,与0.5 mmol/L、1 mmol/L和2 mmol/L处理组均显著降低(P0.05)。结论:1 SIN能抑制胃癌细胞SGC-7901的增殖,且具有时间和浓度依赖性;2 SIN对胃癌细胞SGC-7901中EMT相关蛋白表达和NFκB p65的核转位具有一定的调节作用;3 NFκB p65可能通过核转位来参与SN对SGC-7901的EMT调控作用。
[Abstract]:Aim: to study the effects of sinomenine on SGC-7901 proliferation, invasion and metastasis related genes and protein expression in human gastric cancer cells.Methods the survival rate of human gastric cancer SGC-7901 cells was detected by using different concentrations of SIN.WST-1 assay was used to detect the cell survival rate of 2. 2 MMP expression in human gastric cancer cells treated with different concentrations of SIN. Elisa was used to detect the changes of MMP-2 and MMP-9 content in cell culture medium. 3. 3 RT-PCR detection of SIN in different concentrations of SIN on human gastric cancer SGC-7901 cells.Effect of 4. 4 Western blotting on the expression of EMT protein in SGC-7901 cells of gastric carcinoma; the effect of SIN at different concentrations on the expression of NF 魏 B p65 protein in SGC-7901 cell nucleus of human gastric cancer.Results Human gastric cancer SGC-7901 cells were treated with different concentrations of SIN. The results showed that the survival rate of SGC-7901 cells treated with 1 mmol / L ~ (2 mmol / L) ~ (2.5 mmol/L) and 3mmol/L was significantly lower than that of control group (P _ (0.05)) after 24 h treatment.After 48 h treatment, the cell survival rate of 0.5 mmol / L ~ (-1) mmol / L ~ (2 mmol / L) 2 mmol / L ~ (2 mmol/L) and 3 mmol/L treatment group was significantly lower than that of the control group (P 0.05), and the MMPs content of SGC-7901 cell culture medium was decreased after treatment with sinomenine in a time-and dose-dependent manner.The content of MMP-2 in the culture medium of each SIN treatment group was significantly lower than that of the control group (P0.05T-1 mmol/L and 2 mmol/L). The results showed that the content of MMP-9 decreased with the increase of SIN concentration.The expression of MMP-2 m RNA, protein and MMP-9 mRNA in human gastric cancer SGC-7901 cells were significantly decreased in each treatment group.The expression of MMP-9 protein in 0.5 mmol / L mmol/L and 2 mmol/L groups also decreased significantly compared with the control group, and the Snail gene and protein expression in 0.5 mmol / L 1 mmol / L and 2 mmol/L treatment group was significantly lower than that in the control group, and the E-cadherin gene and protein surface in the 0. 5 mmol / L 1 mmol / L and 2 mmol/L treatment group was significantly lower than that in the control group.The results of P0.05n.4 Western Blotting showed that,The expression of NF 魏 B p65 in SGC-7901 cells decreased with the increase of SIN concentration.Compared with the control group, 0. 5 mmol 路L ~ (-1) L ~ (-1) mmol/L and 2 mmol/L treatment group significantly decreased P 0. 05%.Conclusion 1 SIN can inhibit the proliferation of SGC-7901 in gastric cancer cells.The expression of EMT related protein in SGC-7901 and nuclear translocation of NF 魏 B p65 may be involved in the EMT regulation of SGC-7901 by nuclear translocation of NF 魏 B p65 in gastric cancer cells.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.2

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