软坚消瘤片药物血清对乳腺癌MCF-7细胞生长的机制研究

Mechanism Research of Inhibition of MCF-7 Cell Growth with Ruanjian Xiaoliu Pill Drug Serum

目的:通过观察软坚消瘤片药物血清对乳腺癌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,探讨软坚消瘤片治疗乳腺肿块性疾病的可能机制。方法:制备软坚消瘤片含药动物血清,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24h后能明显抑制乳腺癌细胞株MCF-7的CYP19mRNA、ERmRNA表达水平,表达水平分别降低0.32和0.09倍;药物血清作用48h后S期和G2/M期细胞所占比例降低,尤以G2/M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48h以后细胞生长曲线幅度开始明显低于对照组,在96h时抑制效果最明显。结论:软坚消瘤片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,同时抑制一定时段的细胞生长。

Objective：To investigate the effects of Ruanjian Xiaoliu Pill （RJXLp） drug serum on aromatase, estrogen receptor（ER） expression level, cell generation cycle and apoptosis in breast carcinoma cell line MCF-7 cells. Methods： RJXLp drug serum was prepared and MCF-7 cells were treated with it for 24, 48 and 96h respectively. Then the expression of aromatase and ER mRNA were analyzed by qRT-PCR, cell generation cycle and apoptosis were assayed by flow cytometry（FCM） and cell growth curve was determined by MTS. Results： After incubated in RJXLp drug serum for 24h, expression of CYP19 mRNA、ER mRNA of MCF-7 cells decreased significantly compared to control group （0.32 and 0.09 fold, respectively）. For the cell generation cycle analysis, cell cycle process of MCF-7 cells was changed after 48h treatment with RJXLp drug serum. Cells in S and G2/M phase decreased, especially in G2/M phase, while cells in G0/G1 phase increased. Both early and late apoptosis cells increased compared with control group. MTS assay showed cell growth curve declined after incubated in drug serum for 48h for the drug serum group and inhibition effects reached the peak when incubated for 96h. Conclusion：RJXLp drug serum can inhibit estrogen biosynthesis, attenuate its effects, and suppress cell growth as well.