酸性微环境下胃癌SGC7901细胞自噬水平的变化及其对细胞增殖影响的研究

发布时间：2018-08-24 08:22

【摘要】：目的:探究酸性环境下胃腺癌细胞株SGC7901自噬水平的改变以及这种改变对细胞增殖活性的影响。方法:选择人胃腺癌细胞株SGC7901作为研究细胞;采用透射电镜、激光共聚焦显微镜检测酸应激对胃癌细胞株SGC7901自噬体形成情况的影响;用Western-Blot检测短时间和长时间暴露于酸性条件下培养的SGC7901细胞自噬相关蛋白LC3-II、p62的表达情况,分别探究不同时间长度酸性环境作用下SGC7901细胞自噬水平的变化,并检测巴伐洛霉素A1处理后SGC7901细胞LC3-II、p62表达,探究酸性条件下SGC7901细胞自噬流的变化;通过CCK-8法检测抑制酸性环境下的自噬对细胞增殖活性的影响。结果:1.透射电镜下观察到,与中性环境下培养的SGC7901细胞相比,酸性环境中培养的胃癌SGC7901细胞内自噬体、自噬溶酶体形成增多;激光共聚焦显微镜下观察到酸性条件下培养的SGC7901细胞细胞内LC3荧光颗粒较中性环境下培养的SGC7901细胞细胞内LC3荧光颗粒数量增多,中性环境下培养的SGC7901细胞细胞内LC3荧光弥散分布。2.与中性环境下培养的SGC7901细胞相比,酸性条件下培养的的胃癌SGC7901细胞LC3-II生成增多,p62表达减少;长时间培养于pH 6.5环境下的胃癌SGC7901细胞LC3-II持续高表达,p62持续低表达。3.酸性环境下自噬流增强。4.抑制酸性环境诱导的自噬可引起细胞增殖抑制。结论:胃癌SGC7901细胞在酸性环境中自噬水平上调,且这种自噬在长时间培养时持续稳定存在,不会随时间的延长发生增多或衰减。胃癌SGC7901细胞在酸性环境下较在正常环境下增殖缓慢;在酸性环境和正常环境下抑制自噬均可以引起胃癌SGC7901细胞增殖抑制,酸性环境下的自噬对胃癌SGC7901具有保护性作用。
[Abstract]:Aim: to investigate the changes of SGC7901 autophagy and its effect on proliferation of gastric adenocarcinoma cell line in acidic environment. Methods: human gastric adenocarcinoma cell line SGC7901 was selected as the study cell, and the effect of acid stress on the formation of autophagy in gastric cancer cell line SGC7901 was detected by transmission electron microscope and confocal laser microscope. Western-Blot was used to detect the expression of autophagy associated protein (LC3-II,p62) in SGC7901 cells exposed to acidic conditions for a short time and a long time. The changes of autophagy level of SGC7901 cells were investigated under acidic environment for different periods of time. The expression of LC3-II,p62 in SGC7901 cells was detected after treatment with bavaromycin A1, and the changes of autophagy in SGC7901 cells under acidic conditions were investigated, and the effect of inhibition of autophagy on proliferation of SGC7901 cells in acidic environment was detected by CCK-8 assay. The result is 1: 1. Compared with SGC7901 cells cultured in neutral environment, autophagy and lysosome formation in SGC7901 cells of gastric cancer cultured in acidic environment increased under transmission electron microscope. Laser confocal microscopy showed that the number of LC3 fluorescent particles in SGC7901 cells cultured in acid medium was higher than that in SGC7901 cells cultured in neutral environment. Fluorescence dispersion of LC3 in SGC7901 cells cultured in neutral environment. 2. 2. Compared with SGC7901 cells cultured in neutral environment, the expression of LC3-II increased in SGC7901 cells cultured in acid medium, and the expression of p62 in SGC7901 cells cultured in pH 6.5 for a long time was higher than that in SGC7901 cells cultured in pH 6.5 for a long time. The autophagy was enhanced in acidic environment. Inhibition of acidic environment induced autophagy can induce cell proliferation inhibition. Conclusion: the level of autophagy of gastric cancer SGC7901 cells is up-regulated in acidic environment, and the autophagy exists steadily for a long time and does not increase or decrease with the prolongation of time. The proliferation of gastric cancer SGC7901 cells in acidic environment was slower than that in normal environment, and inhibition of autophagy in acidic environment and normal environment could induce inhibition of SGC7901 cell proliferation. Autophagy in acidic environment had protective effect on gastric cancer SGC7901.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

【相似文献】