通络散结丸药物血清对乳腺癌MCF-7细胞生长的抑制作用

Inhibitory Effects of "Tongluo Sanjie Pill" Drug Serm on MCF-7 Cell Growth of Breast Carcinoma

目的:通过观察通络散结丸药物血清对乳腺癌MCF-7细胞的芳香化酶和雌激素受体基因的表达、细胞增殖周期及细胞凋亡的影响,探讨通络散结丸治疗乳腺肿块性疾病的可能机制。方法:制备通络散结丸含药动物血清,以体外培养的MCF-7细胞为研究对象,以qRT-PCR法检测乳腺癌细胞芳香化酶CYP19及雌激素受体ER mRNA表达水平;流式细胞术(FCM)测定细胞增殖周期和凋亡水平;MTS实验测定细胞生长曲线。结果:与空白对照组比较,含药血清组作用细胞24 h后能明显抑制乳腺癌细胞株MCF-7的CYP19 mRNA、ER mRNA表达水平,表达水平分别降低0.346和0.128倍;药物血清作用48 h后S期和G2/M期细胞所占比例降低,尤以S期最为显著,相应的G0/G1期细胞增加,药物血清抑制了细胞增殖的正常进行;凋亡实验示细胞早期凋亡率和晚期凋亡率均增加,药物血清组总体凋亡率高于对照组;药物血清作用细胞48 h以后细胞生长曲线幅度开始明显低于对照组,在96 h时抑制效果最明显。结论:通络散结丸药物血清影响乳腺癌MCF-7细胞的雌激素合成及其作用发挥,同时抑制细胞生长。

Objective： To investigate the effects of ＂Tongluo Sanjie Pill＂ （TLSJp） drug serum on aromatase and estrogen receptor （ER） gene expression, cell proliferation cycle and apoptosis of MCF-7 cells in breast carcinoma cell line, and explore the mechanisms of TLSJp in treating breast tumor. Methods： TLSJp drug serum was prepared and MCF-7 cells were treated with it for 24, 48 and 96 h respectively. Then the aromatase CYP19 and ER mRNA expressions were detected by qRT-PCR method, cell proliferation cycle and apoptosis were assayed by flow cytometzy （ FCM）, and the cell growth crave was determined by MTS. Results： After incubated in TlS]p drug set＂urn fro＂ 24 b, the expressions of CYP19 mRNA and ER mRNA of MCF-7 cells decreased signifi- cantly compared with blank control group （0. 346 and 0. 128 fold, respectively）. For the cell proliferation cycle analysis, the cell cycle process of MCF-7 cells was changed after 48 h treatment with TLSJp drug serum; the cells in S and G2/M phases decreased, especially in S phase, while the cells in G0/G1 phase increased. The apoptosis experiment showed that both early and late apoptosis cells in drug serum group increased compared with blank contrul group. MTS assay showed that after incubated in drug serum for 48 h the cell growth curve significantly declined compared with blank contrul group, and the inhibitory effects reached the peak when incubated in drug serum for 96 h. Conclusion ： TLSJp drug serum can influence the estrogen biosynthesis and its effects of MCF-7 eells in breast carcinoma, and inhibit the cell growth.