人胃癌组织及细胞株中SLC6A1的表达及生物学活性研究

发布时间：2018-04-20 10:37

本文选题：胃癌组织 + 胃癌细胞株　；参考：《兰州大学》2015年硕士论文

【摘要】：目的：前期研究发现miR-133b在胃癌中低表达,且能够调控下游基因SLC6A1的表达。本课题研究SLC6A1 mRNA在人胃癌及癌旁组织中的表达,SLC6A1 mRNA及蛋白在人胃癌细胞株中的表达,下调SLC6A1在胃癌细胞株中的表达后,对细胞增殖及凋亡的影响。探讨SLC6A1在胃癌发生发展中的作用,为胃癌未来的靶向治疗提供新的参考依据,进一步为胃癌的诊断、预后提供新的思路和理论基础。方法：SLC6A1 mRNA在人胃癌组织和人正常胃粘膜上皮细胞GES-1及人胃癌细胞株AGS、SGC-790、BGC-823、MGC-803中表达的相对水平运用实时荧光定量PCR检测；SLC6A1蛋白在上述细胞中的表达水平运用蛋白印迹(Western Blot)技术检测；通过RNAi瞬时转染技术沉默正常胃粘膜上皮细胞及各胃癌细胞株中SLC6A1的表达后,以CCK-8法检测SLC6A1对各细胞株增殖的影响,并以流式细胞术(FCM)检测SLC6A1对细胞凋亡的影响及其对细胞周期特异性的影响。结果：SLC6A1 mRNA在胃癌组织中的表达较其在癌旁组织中的表达量高,SLC6A1 mRNA及蛋白在人胃癌细胞株中及GES-1细胞株中的表达由高到低依次为SGC-790、BGC-823、MGC-803、AGS、GES-1。si-SLC6A1处理各细胞株后,各细胞株中SLC6A1的表达量下降。下调SLC6A1的表达后,CCK-8检测各细胞株增殖明显受到抑制。流式细胞术检测发现下调SLC6A1的表达后各细胞株的凋亡率有所增加；SLC6A1表达下调后,各细胞周期变化为：S期含量增加,G1期含量降低,细胞被阻滞在S期。结论：1、SLC6A1在胃癌组织中的表达量高于癌旁组织,在胃癌细胞株中的表达量高于在正常胃粘膜上皮细胞株中的表达,SLC6A1可能是一种促癌基因。2、SLC6A1的在胃癌细胞株中的表达在中分化胃癌细胞株SGC-7901中最高,在高分化胃癌细胞株AGS中最低,SLC6A1在胃癌中发挥作用可能与细胞分化过程有关。3、SLC6A1可以促进胃癌细胞增殖,抑制胃癌细胞凋亡并影响细胞周期的变化。
[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.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.2

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