西黄丸含药血清对人乳腺癌细胞株MDA-MB-435和MCF-7细胞增殖的影响

Effect of serum containing Xihuang pill on proliferation of human breast cancer cell line MDA-MB-435 and MCF-7 Cells

研究西黄丸含药血清对人乳腺癌细胞MDA-MB-435和MCF-7细胞增殖以及Bcl-2,Bax,TP53 mRNA及蛋白表达的影响,以期探讨西黄丸抗乳腺癌的作用特点和机制。大鼠灌胃给药制备西黄丸含药血清,实验设置大鼠空白血清组（Control）和西黄丸含药血清组（XH）,以含药血清干预人乳腺癌细胞株MDA-MB-435和MCF-7,MTT法检测细胞增殖,RT-PCR法检测Bcl-2,Bax,TP53 mRNA的表达,免疫荧光法检测Bax蛋白的表达。结果显示,西黄丸大鼠含药血清可抑制MDA-MB-435细胞增殖（P〈0.05）,西黄丸大鼠含药血清可以提高MDA-MB-435细胞中TP53和Bax mRNA以及Bax蛋白的表达（P〈0.05）,降低Bcl-2/Bax的比值（P〈0.05）。同时西黄丸含药血清可以上调MCF-7细胞中Bax mRNA,并降低Bcl-2/Bax的比值（P〈0.05）,而西黄丸含药血清干预MCF-7细胞后TP53 mRNA和Bax蛋白表达无显著性差异。结果表明,西黄丸含药血清可诱导雌激素受体阴性乳腺癌细胞凋亡,机制有可能是通过上调TP53 mRNA表达,从而诱导凋亡蛋白Bax表达,促进Bax向线粒体转移,最终发挥诱导细胞凋亡作用。

To study the effect of serum containing Xihuang pill on the proliferation of human breast cancer cell lines MDA-MB-435 and MCF-7 and the gene and protein expressions of Bcl-2,Bax,TP53,in order to explore the effect and mechanism of Xihuang pill in resisting breast cancer. The serum of the rats was prepared by the method of MTT assay. The expressions of Bcl-2 and Bax were detected by RT-PCR. The serum levels of Bcl-2 and Bax and the mRNA expression of TP53 were detected by immunofluorescence. The rats with serum containing Xihuang pill could inhibit the proliferation of MDA-MB-435 cells and MCF-7 cells（P 0. 05）. The serum containing Xihuang pill increased TP53 and Bax in MDA-MB-435 cells（P 0. 05）,and the ratio of Bcl-2/Bax was decreased（P 0. 05）. Meanwhile,the serum containing Xihuang pill could up-regulate the mRNA expression of Bax in MCF-7 cells and decrease the expression of Bcl（P 0. 05）,but there was no significant difference between the expression of TP53 mRNA and Bax protein expressions after the treatment of MCF-7 cells with Xihuang pill serum. Serum containing Xihuang pill can induce the apoptosis of human breast cancer cells,and the mechanism of estrogen receptor-negative breast cancer cell apoptosis may be induced by up-regulating the mRNA expression of TP53,which can induce the expression of Bax and promote the metastasis of Bax to mitochondria,and ultimately play the role of inducing apoptosis.