奥巴克拉联合5-氟尿嘧啶对低氧性胰腺癌细胞凋亡的作用研究

目的: 研究在低氧条件下，奥巴克拉(obatoclax，OBX)联合5-fluorouracil(5-FU)对胰腺癌HPAC细胞凋亡的影响及其可能的作用机制。方法: 建立低氧条件；采用Western blot检测正常氧及低氧条件下HPAC细胞中HIF-1α抗体的表达；Hoechst 33258染色检测OBX在正常氧及低氧条件下对胰腺癌细胞凋亡的影响；细胞分组为低氧组、5-FU组、OBX+5-FU组，MTT检测各组中细胞增殖活性， Western blot检测各组中HIF-1α、Bcl-2、Bax蛋白的表达。结果: 低氧条件下HIF-1α的表达显著高于其在正常氧条件下的表达，说明低氧条件成功；在低氧条件下，OBX诱导胰腺癌HPAC细胞凋亡的能力增强，凋亡率增加，且呈一定的剂量依赖关系；低氧条件下5-FU可抑制胰腺癌细胞活力，降低细胞中HIF-1α、Bcl-2的表达，促进Bax的表达；OBX联合5-FU组，细胞活力显著降低，HIF-1α、Bcl-2的表达进一步下降，Bax的表达明显升高。结论: 在低氧条件下，OBX联合5-FU可通过抑制HIF-1α和Bcl-2的表达，促进Bax的表达，诱导细胞凋亡而抑制胰腺癌HPAC细胞的增殖。

Effect of obatoclax in combination with 5-fluorouracil on the apoptosis of pancreatic cancer cells under hypoxic

AIM: To explore the effects of OBX combined with 5-FU on pancreatic cancer cell apoptosis (HPAC), and its possible mechanism. METHODS: Hypoxic condition was established. Western blot method was used to detect the expression of HIF-1α in HPAC cells under normal oxygen and hypoxia condition. Hoechest33258 staining was applied to analyze the effects of OBX on apoptosis of HPAC cells under normal oxygen and hypoxia condition. MTT assay was used to detect cell viability in hypoxic group, 5-FU group and OBX combined with 5-FU group. Western blot was applied to detect HIF-1α, Bcl-2 and Bax proteins expression in HPAC cells of hypoxic group, 5-FU group and OBX combined with 5-FU group. RESULTS:Under hypoxia condition, the expression of HIF-1α in HPAC cells was much higher than its expression under normal oxygen. The effect of OBX on HPAC cells apoptosis enhanced with dose-dependent in a certain range of concentrations. Under hypoxia condition, 5-FU decreased pancreatic cancer cells viability and reduced the expressions of HIF-1α, Bcl-2 proteins, but promoted the expression of Bax protein in HPAC cells. OBX combined with 5-FU enhanced the inhibitive effects on cells viability. The expressions of HIF-1α, Bcl-2 proteins reduced further, but the expression of Bax protein increased significantly. CONCLUSION: Under hypoxia condition, OBX combined with 5-FU can enhance the inhibition on HPAC cells viability, and the induction on HPAC cells apoptosis, the mechanism maybe related to reduce the expressions of HIF-1α, Bcl-2 proteins and increasing Bax expression.