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目的:观察舒肝凉血方对雌激素受体阳性乳腺癌MCF-7和T47D细胞增殖的影响,并检测舒肝凉血方对两个细胞系中硫酸酯酶的活性及其mRNA表达的影响。方法:培养雌激素受体阳性的乳腺癌细胞MCF-7和T47D,给予不同浓度的舒肝凉血方处理24 h后,MTT法检测复方对MCF-7和T47D细胞增殖的影响;RT-PCR检测舒肝凉血方对硫酸酯酶mRNA表达的影响。培养MCF-7和T47D细胞,转染萤火虫荧光素酶和海肾荧光素酶质粒,给予不同浓度的舒肝凉血方处理24 h后,双荧光素酶报告基因检测试剂盒检测复方对两个细胞系中硫酸酯酶活性的影响。结果:舒肝凉血方显著抑制了MCF-7和T47D细胞的增殖,并呈现出剂量依赖关系,半数抑制浓度(IC50)分别是46.08 g.L-1和22.98 g.L-1;低浓度的舒肝凉血方对硫酸酯酶mRNA的表达无明显影响,高浓度时则显著下调了硫酸酯酶mRNA的表达;舒肝凉血方显著降低了MCF-7和T47D细胞中硫酸酯酶的活性,且呈现出剂量依赖性。结论:舒肝凉血方能显著抑制MCF-7和T47D细胞的增殖,此作用可能与该方降低硫酸酯酶活性以及下调硫酸酯酶mRNA的表达有关。
Objective: To observe the effect of Shugan Liangxue Prescription on the proliferation of estrogen receptor positive breast cancer MCF-7 and T47D cells and to examine the effect of Shugan Liangxue Prescription on the activity and expression of sulphatase in two cell lines . Methods: The estrogen receptor positive breast cancer cells MCF-7 and T47D were cultured and treated with different concentrations of Shu Gan Liang Xue Decoction for 24 h. MTT assay was used to detect the effect of compound prescription on the proliferation of MCF-7 and T47D cells. RT-PCR Effect of Shugan Liangxue Recipe on the Expression of Sulfatase mRNA. The MCF-7 and T47D cells were cultured and transfected with firefly luciferase and Renilla luciferase plasmids, treated with different concentrations of Shugan Liangji Decoction for 24 h. The dual luciferase reporter gene assay kit was used to test the effect of compound Effect of sulfatase activity in cell lines. Results: Shugan Liangxue Fang significantly inhibited the proliferation of MCF-7 and T47D cells in a dose-dependent manner with IC50 values of 46.08 gL-1 and 22.98 gL-1, respectively. Low concentrations of Shugan Liang Suexuefang had no significant effect on the expression of sulfatase mRNA, but significantly decreased the expression of sulfatase mRNA at high concentration. Shugan Cooling Decoction significantly reduced the activity of sulfatase in MCF-7 and T47D cells Dose-dependent. Conclusion: Shugan Liangxue can significantly inhibit the proliferation of MCF-7 and T47D cells, which may be related to the decrease of sulfatase activity and the expression of sulfatase mRNA.