發(fā)布時(shí)間：2018-08-24 08:22

【摘要】：目的:探究酸性環(huán)境下胃腺癌細(xì)胞株SGC7901自噬水平的改變以及這種改變對(duì)細(xì)胞增殖活性的影響。方法:選擇人胃腺癌細(xì)胞株SGC7901作為研究細(xì)胞;采用透射電鏡、激光共聚焦顯微鏡檢測(cè)酸應(yīng)激對(duì)胃癌細(xì)胞株SGC7901自噬體形成情況的影響;用Western-Blot檢測(cè)短時(shí)間和長(zhǎng)時(shí)間暴露于酸性條件下培養(yǎng)的SGC7901細(xì)胞自噬相關(guān)蛋白LC3-II、p62的表達(dá)情況,分別探究不同時(shí)間長(zhǎng)度酸性環(huán)境作用下SGC7901細(xì)胞自噬水平的變化,并檢測(cè)巴伐洛霉素A1處理后SGC7901細(xì)胞LC3-II、p62表達(dá),探究酸性條件下SGC7901細(xì)胞自噬流的變化;通過CCK-8法檢測(cè)抑制酸性環(huán)境下的自噬對(duì)細(xì)胞增殖活性的影響。結(jié)果:1.透射電鏡下觀察到,與中性環(huán)境下培養(yǎng)的SGC7901細(xì)胞相比,酸性環(huán)境中培養(yǎng)的胃癌SGC7901細(xì)胞內(nèi)自噬體、自噬溶酶體形成增多;激光共聚焦顯微鏡下觀察到酸性條件下培養(yǎng)的SGC7901細(xì)胞細(xì)胞內(nèi)LC3熒光顆粒較中性環(huán)境下培養(yǎng)的SGC7901細(xì)胞細(xì)胞內(nèi)LC3熒光顆粒數(shù)量增多,中性環(huán)境下培養(yǎng)的SGC7901細(xì)胞細(xì)胞內(nèi)LC3熒光彌散分布。2.與中性環(huán)境下培養(yǎng)的SGC7901細(xì)胞相比,酸性條件下培養(yǎng)的的胃癌SGC7901細(xì)胞LC3-II生成增多,p62表達(dá)減少;長(zhǎng)時(shí)間培養(yǎng)于pH 6.5環(huán)境下的胃癌SGC7901細(xì)胞LC3-II持續(xù)高表達(dá),p62持續(xù)低表達(dá)。3.酸性環(huán)境下自噬流增強(qiáng)。4.抑制酸性環(huán)境誘導(dǎo)的自噬可引起細(xì)胞增殖抑制。結(jié)論:胃癌SGC7901細(xì)胞在酸性環(huán)境中自噬水平上調(diào),且這種自噬在長(zhǎng)時(shí)間培養(yǎng)時(shí)持續(xù)穩(wěn)定存在,不會(huì)隨時(shí)間的延長(zhǎng)發(fā)生增多或衰減。胃癌SGC7901細(xì)胞在酸性環(huán)境下較在正常環(huán)境下增殖緩慢;在酸性環(huán)境和正常環(huán)境下抑制自噬均可以引起胃癌SGC7901細(xì)胞增殖抑制,酸性環(huán)境下的自噬對(duì)胃癌SGC7901具有保護(hù)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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

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