|Alternative Title||The Expression of HORMAD2 and the Effects of It on Proliferation and Apoptosis on Gastric Cancer Cell
|Place of Conferral||兰州
|Other Abstract||Objective: 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...|
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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