兰州大学机构库 >学院待认领
HORMAD2在胃癌中的表达及对胃癌细胞增殖
Alternative TitleThe Expression of HORMAD2 and the Effects of It on Proliferation and Apoptosis on Gastric Cancer Cell
侯向红
Thesis Advisor乔梁 ; 周永宁
2015-05-29
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
KeywordHORMAD2 胃癌 增殖 凋亡
Abstract目的:本课题探讨人胃癌组织及胃癌细胞系中HORMAD2的表达;研究下调HORMAD2对正常胃粘膜上皮细胞GES-1和胃癌细胞株SGC-7901、AGS、MGC-803和BGC-823增殖、凋亡及细胞周期的影响。 方法:qRT-PCR技术检测54对人胃癌组织及癌旁组织中HORMAD2的表达水平。通过qRT-PCR和western blot检测正常胃粘膜上皮细胞GES-1和分化程度不同的胃癌细胞中HORMAD2的相对表达水平。然后用针对HORMAD2合成的特异性的siRNA对细胞株通过脂质体进行瞬时转染,分别在mRNA和蛋白水平检测转染后细胞株HORMAD2的表达情况。使用CCK-8检测下调HORMAD2对GES-1和胃癌细胞株增殖能力的影响;流式细胞术检测细胞通过siRNA转染48h后的细胞周期及细胞凋亡率。 结果:HORMAD2在胃癌组织中的表达量高于癌旁组织(P<0.05);在未分化的胃癌细胞株MGC-803中的表达水平比正常胃粘膜上皮细胞和其他分化类型的胃癌细胞株明显增高(P<0.05),而细胞株SGC-7901和AGS与GES-1相比HORMAD2的表达水平降低(P<0.05)。CCK-8检测到HORMAD2表达下调后正常胃粘膜上皮细胞及胃癌细胞的增殖活性均受到抑制(P<0.05)。而且,下调HORMAD2的表达后GES-1及SGC-7901、AGS、MGC-803和BGC-823的细胞凋亡率明显增高,这种效应尤其在高表达HORMAD2的MGC-803细胞中尤为明显;同时下调HORMAD2使得细胞周期阻滞在了S期,G2/M期停滞。 结论:CT基因 HORMAD2作为一种潜在的癌基因可能参与了胃癌的发生发展。
Other AbstractObjective: We search the expression of HORMAD2 in human gastric cancer tissues and cell lines. Furthermore, we tested the effects of down-regulation HORMAD2 on apoptosis, proliferation and cell cycle on gastric cancer cell SGC-7901, BGC-823, MGC-803 and AGS and the normal human gastric epithelial cell line GES-1.Method:We investigated the expression of HORMAD2 in total of fifty-four pairs of human gastric cancer and adjacent non-tumor tissues and the human gastric cancer cell lines SGC-7901, BGC-823, MGC-803 and AGS and the normal human gastric epithelial cell line GES-1 by qRT-PCR. At the same time, we tested the expression of HORMAD2 protein in those cell lines through western blot. And then, the cell lines (SGC-7901, BGC-823, MGC-803, AGS and GES-1) transfected with siRNA were performed by LipofectamineTM 2000 (Invitrogen) in accordance with the manufacturer’s instructions. The transfection efficiency was identified with qRT-PCR and western blot after 48h and 72h, respectively. The influence of down-regulation HORMAD2 on cell viability was measured by the Cell Counting Kit-8 (CCK-8). Meanwhile, we analyzed the influence of down-regulation of HORMAD2 on cell cycle and cell apoptosis by flow cytometry. Results:We found that the relative expression level of HORMAD2 were significantly up-regulated in gastric cancer specimens compared with the paired non-tumor tissues through qRT-PCR test (P<0.05). The relative level expression of HORMAD2 was meaningfully higher in MGC-803 cells than that of others (P<0.05). However, the relative expression level of HORMAD2 in cell lines AGS and SGC-7901 was significantly lower compared with GES-1 (P<0.05).After two siRNAs specific against HORMAD2 were transfected into GES-1 and gastric cancer cells AGS, SGC-7901, BGC-823 and MGC-803 by LipofectamineTM 2000, the protein level and mRNA level of HORMAD2 was markedly suppressed. We used the Cell Counting Kit-8 (CCK-8) to measure proliferation of cell. The results indicated that the down-regulation of HORMAD2 could dramatically suppress cell growth compared with the negative control siRNA_NC in all cells. After transfected all cells with siRNA_128, siRNA_367 and siRNA_NC 48h, we confirmed that the apoptosis rate was obvious increasing transfected with siRNA_128 and siRNA_367 compared to the negative control siRNA_NC through flow cytometry. Furthermore, HORMAD2 down-regulation could cause an arrest in S phase and blocked cells into G2/M phase of cell cycle.Conclusion:In c...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/224095
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
侯向红. HORMAD2在胃癌中的表达及对胃癌细胞增殖[D]. 兰州. 兰州大学,2015.
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