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目的探讨缺氧状态下及缺氧诱导因子-1α(Hypoxia-inducible factor-1α,HIF-1α)RNA干扰后,HIF-1α的表达及其与胃癌细胞黏附、游走、侵袭能力的相关性。方法将HIF-1α shRNA真核表达质粒pGPU6/GFP/Neo-HIF-1α-2484转染胃癌细胞SGC-7901,构建靶向干扰HIF-1α表达的SGC-7901稳定转染细胞株;采用Western blot法检测缺氧及HIF-1αRNA干扰对SGC-7901细胞HIF-1α表达的影响;MTT法检测缺氧及HIF-1αRNA干扰对SGC-7901细胞黏附能力的影响;采用Boyden Chamber膜侵袭系统,观察缺氧及HIF-1αRNA干扰对SGC-7901细胞游走及侵袭能力的影响。结果 SGC-7901细胞在缺氧培养4、8和16h时,HIF-1α均呈阳性表达,并随时间的延长而显著增加;细胞粘贴率、游走和侵袭穿膜细胞数分别随时间延长而显著增加,与HIF-1α表达呈显著正相关。HIF-1αRNA干扰后,缺氧SGC-7901细胞HIF-1α表达显著下降,细胞粘贴率、游走和侵袭穿膜细胞数均显著下降。结论 HIF-1α的过度表达导致胃癌细胞黏附、游走及侵袭能力增强,可能是胃癌局部侵袭和远处转移的重要原因之一。
Objective To investigate the expression of HIF-1α in hypoxia and Hypoxia-inducible factor-1α (HIF-1α) RNA interference and its relationship with adhesion, migration and invasion of gastric cancer cells. Methods SGC-7901 cells transfected with SGC-7901 cells were transfected with SGC-7901 gastric cancer cells by transfecting HIF-1α shRNA eukaryotic expression vector pGPU6 / GFP / Neo-HIF-1α-2484. Western Blot The effect of hypoxia and HIF-1αRNA interference on the expression of HIF-1α in SGC-7901 cells was detected by MTT assay. The effect of hypoxia and HIF-1α RNA interference on the adhesion of SGC-7901 cells was detected by Boyden Chamber invasion system. Effects of oxygen and HIF-1α RNA interference on migration and invasion of SGC-7901 cells. Results The expression of HIF-1α in SGC-7901 cells was significantly increased at 4, 8 and 16 h hypoxia, and significantly increased with time. The cell attachment rate, cell migration and invasion cell number increased with time Significantly increased, with a significant positive correlation with HIF-1αexpression. After HIF-1α RNA interference, the expression of HIF-1α in hypoxia SGC-7901 cells was significantly decreased, and the cell attachment rate, migration and invasion cell numbers were significantly decreased. Conclusion The overexpression of HIF-1α leads to the enhancement of adhesion, migration and invasion of gastric cancer cells, which may be one of the important reasons for the local invasion and distant metastasis of gastric cancer.