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目的观察不同氧浓度下结肠癌细胞株SW480中缺氧诱导因子(hypoxia inducible factor HIF)-1α、糖酵解限速酶表达的差异及细胞培养上清液中乳酸浓度的变化。方法将传代培养的结肠癌细胞置于不同氧浓度的环境中培养12小时,采用反转录-聚合酶链式反应(RT-PCR)和western-blot法检测HIF-1αmRNA和蛋白表达水平;RT-PCR和western-blot法检测常氧和缺氧状态下结肠癌细胞糖酵解限速酶葡萄糖转运蛋白(glucose transporter 1,GLUT-1)、乳酸脱氢酶A(lactate dehydrogenase A,LDH-A)基因的表达。比色法测定结肠癌细胞上清液中的乳酸浓度。结果随着培养环境氧浓度下降,结肠癌细胞中HIF-1α表达逐步增加(P<0.05),糖酵解限速酶GLUT-1和LDH-A表达逐步增加(P<0.05),结肠癌细胞培养上清液中乳酸浓度逐步增高(P<0.05)。结论低氧环境可以上调结肠癌细胞中HIF-1α和GLUT-1、LDH-A的表达,以及细胞培养上清液中乳酸浓度。随着氧浓度下降,差异有统计学意义(P<0.05)。低氧环境下HIF-1α上调可促进结肠癌细胞的增殖。
Objective To observe the difference of hypoxia inducible factor - 1α (HIF - 1α), rate - limiting enzyme (GLP) and the lactate concentration in cell culture supernatant of colon cancer cell line SW480 under different oxygen concentrations. Methods Cultured colon cancer cells were cultured in different oxygen concentrations for 12 hours. The mRNA and protein levels of HIF-1α were detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. RT The expressions of glucose transporter 1 (GLUT-1), lactate dehydrogenase A (LDH-A) in colon cancer cells under normoxia and hypoxia were detected by PCR and western-blot. ) Gene expression. Colorimetric assay of lactate concentration in colon cancer cell supernatant. Results The expression of HIF-1α in colon cancer cells gradually increased (P <0.05) and the rate of GLUT-1 and LDH-A in the culture medium increased gradually The concentration of lactate in the culture supernatant gradually increased (P <0.05). Conclusion Hypoxia can up-regulate the expression of HIF-1α, GLUT-1 and LDH-A in colon cancer cells and the concentration of lactate in cell culture supernatant. With the oxygen concentration decreased, the difference was statistically significant (P <0.05). Hypoxia HIF-1α upregulation can promote the proliferation of colon cancer cells.