清癌汤体外抗肿瘤作用及其机制研究

Study on Antitumor Activities and Its Mechanism of Qing-Ai Decoction In Vitro

目的:探讨清癌汤体外对肿瘤细胞的增殖、凋亡及p53表达的影响。方法:倒置显微镜下观察加药后细胞形态的变化,MTI法检测细胞增殖,流式细胞仪Annexin V-FITC/PI双染法检测细胞凋亡情况,JC-1检测细胞膜电位变化情况,免疫荧光法检测细胞内p53蛋白表达情况。结果:清癌汤在高浓度作用下,肿瘤细胞形态发生了明显变化,对人乳腺癌细胞(MCF-7)、人肝癌细胞(HepG2)、人胃癌细胞(MGC-803)具有明显的抑制作用,并呈浓度依赖性,而对大鼠肾上腺嗜铬瘤细胞(PC12)无明显抑制作用。清癌汤可诱导MGC-803发生凋亡并呈明显的浓度依赖性,降低线粒体膜电位,从而引起细胞凋亡,并能升高p53蛋白的表达量。结论:清癌汤具有良好的体外抗肿瘤作用,且无明显神经毒性,该作用与其诱导细胞凋亡包括降低线粒体膜电位和上调p53蛋白表达有关。

This article was aimed to study the effects of Qing-Ai（QA） decoction on cell proliferation,apoptosis and expression of p53.The morphological changes of cancer cells were observed after drug administration by inverted microscope.The cell proliferation was detected by MTT assay.Annexin V-FITC/PI double staining was used to test apoptosis of cells by the flow cytometry.JC-1 was used to detect the electric potential changes of cell membranes.Immunofluorescence method was used to detect the expression quantity of p53 protein in cells.The results showed that under high concentration of QA decoction,there were obvious changes on cell morphologyIt exerted obvious inhibition on the proliferation of MCF-7,HepG2 and MGC-803 cells.Besides,the inhibition rates grew as the concentration grew.However,it had no influence on PC12.Furthermore,QA decoction induced the apoptosis of MGC-803 with obvious concentration-dependent,reduced the membrane potential of mitochondrion.It also increased the expression of p53 protein.It was concluded that QA decoction had good antitumor effects in vitro.There was no obvious neurotoxicity.Its mechanism was related to its inducing of cell apoptosis,which included the reducing of membrane potential of mitochondrion and upregulating of expression of p53 protein.