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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 effect of Tongluosanjie pill on the expression of aromatase and estrogen receptor gene, cell cycle and apoptosis in breast cancer MCF-7 cells, Possible mechanism of sexually transmitted diseases. Methods: MCF-7 cells cultured in vitro were prepared and the expression levels of CYP19 and estrogen receptor ER mRNA were detected by qRT-PCR. Flow cytometry Cytometry (FCM) was used to determine the cell cycle and the level of apoptosis. MTS assay was used to determine the cell growth curve. RESULTS: Compared with the blank control group, the effect of the drug-containing serum for 24 h significantly inhibited the expression of CYP19 mRNA and ER mRNA in breast cancer cell line MCF-7 by 0.346 and 0.128 times, respectively. The proportion of cells in the late S phase and the G2 / M phase decreased, especially in the S phase. The corresponding G0 / G1 phase cells increased, and the drug serum inhibited the normal cell proliferation. The apoptosis rate showed the early apoptosis rate and The late apoptosis rate increased, and the overall apoptosis rate of the drug serum group was higher than that of the control group. After 48 h, the cell growth curve range of drug serum started to be significantly lower than that of the control group, and the inhibition effect was the most obvious at 96 h. Conclusion: Tongluosanjie pill serum can affect estrogen synthesis and its function in breast cancer MCF-7 cells and inhibit cell growth.