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目的:观察曲古抑菌素A(TSA)体外对结肠癌HT-29细胞凋亡及低氧状态下其缺氧诱导因子(HIF)-1α表达的影响。方法:体外培养结肠癌HT-29细胞,给予TSA 100、200、400、600、800 nmol/L分别处理12、24、48、72 h,对照组加入同等体积的DMSO。MTT法检测各组HT-29细胞的存活率;Annexin V、TUNEL法检测HT-29细胞凋亡率;RT-PCR、免疫印迹法分别检测低氧状态下(37℃、1%O_2)HT-29细胞HIF-1αmRNA及蛋白的表达。结果:MTT法检测结果发现.与对照组相比,TSA处理组HT-29细胞活力明显降低(P<0.05),随着浓度的增大、时间的延长.其抑制作用逐渐增强。Annexin V、TUNEL法检测结果发现.TSA能诱导HT-29细胞的早期凋亡.TSA(200、400、600、800 nmol/L)处理组凋亡率明显高于对照组(P<0.05)。低氧条件下,TSA抑制HT-29细胞HIF-1α蛋白的表达,随着浓度的增大,其抑制作用逐渐增强;而对HIF-1αmRNA的表达无影响。结论:TSA体外可能通过抑制低氧条件下HT-29细胞HIF-1α蛋白的表达.诱导细胞凋亡.抑制细胞增殖。
Objective: To observe the effect of trichostatin A (TSA) on the apoptosis of HT-29 colon carcinoma cells and the hypoxia inducible factor (HIF) -1α expression in HT-29 cells in vitro. Methods: HT-29 cells were cultured in vitro and treated with TSA at 100, 200, 400, 600 and 800 nmol / L for 12, 24, 48 and 72 h, respectively. The control group was given the same volume of DMSO. The survival rate of HT-29 cells was determined by MTT assay. The apoptosis rate of HT-29 cells was detected by Annexin V and TUNEL method. The expression of HT- 29 cells HIF-1αmRNA and protein expression. Results: The results of MTT assay showed that compared with the control group, the viability of HT-29 cells in TSA-treated group was significantly decreased (P <0.05), and the inhibitory effect was gradually increased with the increase of concentration and time. The result of Annexin V and TUNEL showed that TSA could induce the early apoptosis of HT-29 cells, and the apoptotic rate of TSA (200,400,600,800 nmol / L) group was significantly higher than that of the control group (P <0.05). Under hypoxic conditions, TSA inhibited the expression of HIF-1αprotein in HT-29 cells. With the increase of concentration, TSA inhibited the expression of HIF-1αprotein gradually, but had no effect on the expression of HIF-1αmRNA. Conclusion: TSA can inhibit the hypoxia-induced HIF-1α expression in HT-29 cells, induce apoptosis and inhibit cell proliferation.