过表达CDX1蛋白对直肠癌细胞增殖及能量代谢的影响及机制研究

Effects and Its Mechanism of Overexpression of CDX1 on Proliferation and Energy Metabolism of Rectal Cancer Cells

目的:探讨过表达尾侧同源盒转录因子1(CDX1)蛋白对直肠癌细胞增殖及能量代谢的影响。方法:以直肠癌细胞SW117为研究对象,细胞转染pIRES(空载体组)、pIRES-CDX1(过表达组),同时设置对照组(只加入转染试剂)。培养48 h后,Western blot检测细胞中CDX1、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、蛋白激酶B(Akt)、磷酸化蛋白激酶B(p-Akt)表达水平,噻唑蓝(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,试剂盒检测细胞中三磷酸腺苷(ATP)含量、丙酮酸激酶活性、己糖激酶活性及培养液上清中乳酸含量。结果:空载体组细胞中CDX1、Cleaved Caspase-3、Akt、p-Akt表达水平及细胞存活率、凋亡率、ATP含量、丙酮酸激酶活性、己糖激酶活性及培养液上清中乳酸含量与对照组相比均没有明显差异。过表达组细胞存活率、p-Akt水平、ATP含量、丙酮酸激酶活性、己糖激酶活性及培养液上清中乳酸含量与对组相比均明显下降,凋亡率及细胞中Cleaved Caspase-3表达水平均明显高于对照组。结论:过表达CDX1蛋白能够促进直肠癌细胞凋亡,抑制直肠癌细胞增殖,影响直肠癌细胞能量代谢,作用机制可能与p-Akt水平有关。

Objective： Effects of over expression of CDX1 protein on proliferation and energy metabolism of rectal cancer cells.Methods： Rectal cancer cell line SW117 was used as the research object, the cells were transfected with pIRES（empty vector group） and p IRES-CDX1（over expression group）, meanwhile, the control group（only adding transfection reagent） was set up. After culturing 48 h,Western blot was used to detect the expression levels of CDX1, Cleaved Caspase-3, Akt and p-Akt in the cells, cell proliferation was detected by MTT, cell apoptosis was detected by flow cytometry, kit for detecting ATP content of cells, pyruvate kinase activity, hexokinase activity and lactic acid content in the supernatant. Results： The empty vector group in cells of CDX1, Cleaved Caspase-3, Akt, p-Akt expression level and cell survival rate, apoptosis rate, ATP content, pyruvate kinase activity, hexokinase activity, and lactic acid content in the supernatant compared with the control group had no significant difference. Over expression group cell survival rate, p-Akt level,ATP content, pyruvate kinase activity, hexokinase activity and lactate levels in the supernatant fluids in the group decreased significantly compared with the control group. The apoptosis rate and the expression of Cleaved Caspase-3 in the cells were significantly higher than those in the control group. Conclusion： Overexpression of CDX1 protein can promote apoptosis of rectal cancer cells, inhibition of rectal cancer cell proliferation, disturbance of energy metabolism in rectal cancer cells, the mechanism may be related to p-Akt level.