發(fā)布時(shí)間：2018-04-20 10:37

  本文選題：胃癌組織 + 胃癌細(xì)胞株�。� 參考：《蘭州大學(xué)》2015年碩士論文

【摘要】：目的：前期研究發(fā)現(xiàn)miR-133b在胃癌中低表達(dá),且能夠調(diào)控下游基因SLC6A1的表達(dá)。本課題研究SLC6A1 mRNA在人胃癌及癌旁組織中的表達(dá),SLC6A1 mRNA及蛋白在人胃癌細(xì)胞株中的表達(dá),下調(diào)SLC6A1在胃癌細(xì)胞株中的表達(dá)后,對(duì)細(xì)胞增殖及凋亡的影響。探討SLC6A1在胃癌發(fā)生發(fā)展中的作用,為胃癌未來的靶向治療提供新的參考依據(jù),進(jìn)一步為胃癌的診斷、預(yù)后提供新的思路和理論基礎(chǔ)。方法：SLC6A1 mRNA在人胃癌組織和人正常胃粘膜上皮細(xì)胞GES-1及人胃癌細(xì)胞株AGS、SGC-790、BGC-823、MGC-803中表達(dá)的相對(duì)水平運(yùn)用實(shí)時(shí)熒光定量PCR檢測(cè)；SLC6A1蛋白在上述細(xì)胞中的表達(dá)水平運(yùn)用蛋白印跡(Western Blot)技術(shù)檢測(cè)；通過RNAi瞬時(shí)轉(zhuǎn)染技術(shù)沉默正常胃粘膜上皮細(xì)胞及各胃癌細(xì)胞株中SLC6A1的表達(dá)后,以CCK-8法檢測(cè)SLC6A1對(duì)各細(xì)胞株增殖的影響,并以流式細(xì)胞術(shù)(FCM)檢測(cè)SLC6A1對(duì)細(xì)胞凋亡的影響及其對(duì)細(xì)胞周期特異性的影響。結(jié)果：SLC6A1 mRNA在胃癌組織中的表達(dá)較其在癌旁組織中的表達(dá)量高,SLC6A1 mRNA及蛋白在人胃癌細(xì)胞株中及GES-1細(xì)胞株中的表達(dá)由高到低依次為SGC-790、BGC-823、MGC-803、AGS、GES-1。si-SLC6A1處理各細(xì)胞株后,各細(xì)胞株中SLC6A1的表達(dá)量下降。下調(diào)SLC6A1的表達(dá)后,CCK-8檢測(cè)各細(xì)胞株增殖明顯受到抑制。流式細(xì)胞術(shù)檢測(cè)發(fā)現(xiàn)下調(diào)SLC6A1的表達(dá)后各細(xì)胞株的凋亡率有所增加；SLC6A1表達(dá)下調(diào)后,各細(xì)胞周期變化為：S期含量增加,G1期含量降低,細(xì)胞被阻滯在S期。結(jié)論：1、SLC6A1在胃癌組織中的表達(dá)量高于癌旁組織,在胃癌細(xì)胞株中的表達(dá)量高于在正常胃粘膜上皮細(xì)胞株中的表達(dá),SLC6A1可能是一種促癌基因。2、SLC6A1的在胃癌細(xì)胞株中的表達(dá)在中分化胃癌細(xì)胞株SGC-7901中最高,在高分化胃癌細(xì)胞株AGS中最低,SLC6A1在胃癌中發(fā)揮作用可能與細(xì)胞分化過程有關(guān)。3、SLC6A1可以促進(jìn)胃癌細(xì)胞增殖,抑制胃癌細(xì)胞凋亡并影響細(xì)胞周期的變化。
[Abstract]:Aim: to investigate the low expression of miR-133b in gastric cancer and its ability to regulate the expression of downstream gene SLC6A1. The purpose of this study was to investigate the expression of SLC6A1 mRNA in human gastric carcinoma and its adjacent tissues. The expression of SLC6A1 mRNA and protein in human gastric cancer cell line was studied. After down-regulating the expression of SLC6A1 in gastric cancer cell line, the expression of SLC6A1 mRNA and protein on the proliferation and apoptosis of gastric cancer cell line was studied. To explore the role of SLC6A1 in the development of gastric cancer, to provide a new reference for the future targeted therapy of gastric cancer, and to provide new ideas and theoretical basis for the diagnosis and prognosis of gastric cancer. Methods the relative level of mRNA expression in human gastric carcinoma tissue and normal gastric mucosa epithelial cell line GES-1 and human gastric cancer cell line AGSC-790 BGC-823MGC-803 was detected by real-time fluorescence quantitative PCR. Western blot technique was used to detect the expression level of SLC6A1 protein in these cells. The expression of SLC6A1 in normal gastric mucosal epithelial cells and gastric cancer cell lines was silenced by RNAi transient transfection technique. The effect of SLC6A1 on proliferation of gastric cancer cell lines was detected by CCK-8 assay. Flow cytometry (FCM) was used to detect the effect of SLC6A1 on apoptosis and cell cycle specificity. Results the expression of mRNA in gastric cancer tissues was higher than that in adjacent tissues. The expression of SLC6A1 mRNA and protein in human gastric cancer cell line and GES-1 cell line was in order of SGC-790 BGC-823MGC-823MGC-803AGSGC-803AGSGES-1.si-SLC6A1, the expression of SLC6A1 in each cell line decreased. After down-regulating the expression of SLC6A1, CCK-8 assay showed that the proliferation of all cell lines was significantly inhibited. Flow cytometry showed that after down-regulating the expression of SLC6A1, the apoptosis rate of all cell lines increased. After down-regulation of SLC6A1 expression, the cell cycle changed as follows: the content of cell cycle increased, the content of G _ 1 phase decreased and the cell was blocked in S phase. Conclusion the expression of SLC6A1 in gastric cancer is higher than that in paracancerous tissue. The expression of SLC6A1 in gastric cancer cell line was higher than that in normal gastric mucosal epithelial cell line. The expression of SLC6A1 in gastric cancer cell line may be the highest in the moderately differentiated gastric cancer cell line SGC-7901. The role of SLC6A1 in gastric cancer cell line AGS may be related to the process of cell differentiation. It is suggested that SLC6A1 can promote the proliferation of gastric cancer cells, inhibit the apoptosis of gastric cancer cells and influence the change of cell cycle.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.2

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1 趙秋燕;人胃癌組織及細(xì)胞株中SLC6A1的表達(dá)及生物學(xué)活性研究[D];蘭州大學(xué);2015年

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本文編號(hào)：1777414