左金方和5-FU对胃癌细胞生长及BAX/Bcl-2表达的调控研究

Regulation of Growth and BAX/Bcl-2 Expression of Human Gastric Cancer Cells by Zuojin Decoction and 5-FU

目的:研究左金方和5-FU单独与联合对人正常胃黏膜上皮细胞GES-1和人胃癌MGC803细胞生长及凋亡的影响及对BAX/Bcl-2蛋白表达水平的改变。方法:将不同浓度的左金方、5-FU及两药联合作用于体外培养的人胃黏膜上皮细胞GES-1和人胃癌细胞MGC803,Cell Counting Kit-8(CCK-8)检测细胞增殖率的变化,流式细胞仪检测细胞凋亡,Western blot法检测bcl-2、bax蛋白表达。结果:1.00 ug·mL^(-1)左金方、50.00、100.00 ug·mL^(-1)5-FU作用GES-1细胞24 h后,细胞存活率分别为(107.54±2.25)%、(53.67±1.91)%和(47.43±1.35)%,作用MGC803细胞24 h后,细胞凋亡率分别为(45.46±1.66)%、(37.02±1.68)和(45.60±1.14)%;左金方联合5-FU(1.00+50.00)、左金方联合5-FU(1.00+100.00)作用GES-1细胞24 h后,细胞存活率分别为(81.66±2.22)%和(78.68±2.10)%,作用MGC803细胞24 h后,相应的凋亡率为(61.30±1.71)%和(71.30±1.79)%,与单独用药比较差异有统计学意义。Western Blot凝胶电泳图可以看出,左金方联合5-FU(1.00+50.00)、左金方联合5-FU(1.00+100.00)作用MGC803细胞24 h后,进一步促进Bax的表达,降低Bcl-2的表达,与各单独用药组比较差异有统计学意义。结论:左金方与5-FU联合用药有大幅降低5-FU细胞毒性作用,显著提高人正常胃黏膜上皮细胞GES-1的存活率;通过促进MGC803细胞中Bax蛋白的高表达和Bcl-2的低表达,促进人胃癌细胞的凋亡。

Objective： to study the cell growth of Human Gastric Mucosal Epithelial Cell GES-1,the apoptosis of Human Gastric Carcinoma MGC803 and the changes of Bax / Bcl-2 in the level of protein expression caused by ZuoJinFang and 5-Fu separately and together. Methods： Human Gastric Mucosal Epithelial Cell GES-1 and Human Gastric Carcinoma MGC803 cells were cultured in vitro and treated by ZuoJinFang and 5-Fu with different concentration separately and together,the growth inhibitions of GES-1 cell were analyzed by Cell Counting Kit-8（ CCK-8）,the apoptosis rates of MGC803 cell were studied by flow cytometry and the expressions of Bcl-2 and Bax protein were detected by Western blot.Results： After the cells treated by 1. 00 ug·mL^（-1）ZuoJinFang,50. 00、100. 00 ug·mL^（-1）5-FU for 24 h separately,the percentage of GES-1 cell viabilities were（ 107. 54 ± 2. 25） %,（ 53. 67 ± 1. 91） and（ 47. 43 ± 1. 35） %,the percentages of apoptosis of MGC803 cell were（ 45. 46 ± 1. 66） % 、（ 37. 02 ± 1. 68） and（ 45. 60 ± 1. 14） %; after the cells treated by ZuoJinFang combined with 5-FU（ 1. 00 ＋ 50. 00 or 1. 00 ＋ 100. 00 ug·mL^（-1）） for 24 h,the percentage of GES-1 cell viabilities were（ 81. 66 ± 2. 22） % and（ 78. 68 ± 2. 10） %,the percentages of apoptosis of MGC803 cell were（ 61. 30 ±1. 71） and（ 71. 30 ± 1. 79） %,the differences in comparison with ZuoJinFang and 5-Fu group separately were statistically significant. The Western Blot gel electrophoresis figures show that the expressions of Bax were further increased and the expressions of Bcl-2 were decreased accordingly after the MGC803 cells treated by ZuoJinFang combined with 5-FU（ 1. 00＋ 50. 00 or 1. 00 ＋ 100. 00 ug·mL^（-1）） for 24 h,the differences in comparison with ZuoJinFang and 5-Fu group separately were statistically significant. Conclusion： ZuoJinFang combined with 5-FU,greatly reduced the side effects of 5-FU,significantly increased the survival rates of the GES-1 cells,promoted the apoptosis of gastric carcinoma MGC803 cells by promoting high expression of Bax protein and low expression of Bcl-2 protein.