豆根管食通口服液对人食管癌细胞增殖、凋亡及凋亡相关基因表达的影响

The effect of Dougenguanshitong Oral Liquid on proliferation, apoptosis and relevant gene expressions in esophageal carcinoma cell line

目的:观察中药复方制剂——豆根管食通口服液对体外培养的食管癌Eca-109细胞的增殖抑制、诱导凋亡作用及对凋亡相关基因蛋白表达的影响。方法:随机取Eca-109细胞分为三组:实验组、阳性对照组和空白对照组,采用MTT分析法测定不同浓度的豆根管食通口服液对Eca-109细胞增殖的抑制作用;通过倒置显微镜、HE染色观察凋亡细胞;采用免疫组织化学法测定Eca-109细胞内Bcl-2和Bax蛋白的表达。结果:MTT结果显示,豆根管食通口服液对Eca-109细胞的增殖有明显的抑制作用。倒置显微镜、光学显微镜下可发现药物作用组出现细胞核固缩、核碎裂等。免疫组织化学结果显示药物作用组bcl-2蛋白表达降低,bax蛋白表达增高(P<0.01)。结论:豆根管食通口服液对人食管癌Eca-109细胞系有抗增殖及诱导凋亡作用,其分子机制可能是下调bcl-2基因表达,上调bax基因表达。

Objective： To observe the effect of a combination of traditional Chinese herbs Dougenguanshitong Oral Liquid （DOL） on inhibiting proliferation, inducing apoptosis and regulating relevant gene expressions in human Eca-109 cells. Methods： Eca-109 cells were randomly divided into three groups： experimental group, positive control group and normal control group, MTT assay was used to examine the inhibition by different concentrations of DOL on Eca-109 cells; the cellular morphology of apoptosis was observed by invert microscope and HE staining; Immunohisto-chemistry （IHC） technique was used to measure the expression of Bcl-2 and Bax protein in cells. Results： DOL could inhibit the prolifera- tion of Eca-109 cells according to the result of MTT assay. Chromatin condensation and nuclear fragmentation were ob- served in cells dealt with DOL by microscope. Measured with IHC, the expression of Bcl-2 protein in the cells dealt with DOL was decreased, whereas the expression of Bax protein was increased （P〈0.01）. Conclusion： DOL has the effect of anti-proliferation and can induce apoptosis of Eca-109 cells. The molecular mechanism of DOL for inhibiting Eca-109 cells may be correlated with down-regulating the expression of bcl-2 gene and up-regulating the expression of bax gene.