柴桂龙牡汤含药血清对人乳腺癌细胞MCF-7的抑制作用及对雌激素受体α表达的影响

Effect of Serum Containing Chaiguilongmu Decoction on the Growth Inhibition of Human Breast Cancer MCF-7 Cell and Expression of Estrogen Receptor α

目的观察柴桂龙牡汤(CGLM)含药血清对人乳腺癌MCF-7细胞株体外生长的抑制作用及对雌激素受体α(ERα)表达的影响。方法将人乳腺癌MCF-7细胞株细胞分别加入相应(含药)培养基,对照组含有10%空白大鼠血清,给药组分别含有柴桂龙牡汤低、中、高剂量的10%含药血清。加药后,置37℃及5%CO2培养箱中继续培养48 h。噻唑蓝(MTT)法检测柴桂龙牡汤各组含药血清对人乳腺癌MCF-7细胞增殖的抑制作用。将10%的含药血清培养基加入MCF-7细胞中,孵育48 h,提取细胞总mRNA及蛋白,反转录合成c DNA,采用实时荧光定量PCR方法检测各组细胞中ERαmRNA的表达水平,采用Western blot方法检测各组ERα蛋白表达水平。结果 MTT法检测结果显示,与对照组相比,低、中、高剂量柴桂龙牡汤均可抑制MCF-7细胞体外增殖,中、高剂量组的差异有统计学意义(P<0.01);PCR法检测结果显示,与对照组相比,柴桂龙牡汤低、中、高剂量组中ERαmRNA均有升高,其中高剂量组有显著差异(P<0.01);Western blot法检测结果显示,与对照组相比,低、中、高剂量柴桂龙牡汤均使ERα蛋白表达水平升高,中、高剂量组有显著差异(P<0.01)。结论柴桂龙牡汤对MCF-7细胞的体外生长有抑制作用,并与剂量浓度呈正相关,其机制可能与提高ERαmRNA含量、上调ERα蛋白的表达水平有关。

Objective To observe the inhibitory effect of the serum containing Chaiguilongmu（CGLM） Decoction on the growth inhibition of human breast cancer MCF-7 cell and expression of Estrogen receptor α（ERα）. Methods The human breast cancer MCF-7 cell lines were respectively added into the corresponding（drug-containing） culture medium,in which the control group contained 10%empty rat serum,the administration group contained 10 % of the CGLM serum of low,medium and high dosage group,respectively. After administration,they would be at 37 ℃ and 5% CO2 incubator for continuing to develop in 48 h. MTT assay was used to detect the inhibitory effect of CGLM serum of low,medium and high dosage group on human breast cancer MCF-7 cell proliferation. The serum medium containing 10% CGLM was added into MCF-7 cells,incubation after 48 h,the total mRNA and protein from the cells were extracted,and c DNA was synthesized by reverse transcription. The expression level of ERα mRNA in each group was detected by real-time fluorescence quantitative（PCR） method. The expression level of ERα protein in each group was detected by Western blot method. Results MTT results showed that compared with the control group,the CGLM low,medium and high dose group could inhibit the proliferation in vitro of MCF-7 cells,the difference between the medium and high dose groups was statistically significant（P〈0. 01）. PCR results showed that compared with the control group,the levels of ERα mRNA in the CGLM low,medium and high dose group were increased,among which the high-dose group was significantly different（P〈0. 01）. Western blot results showed that compared with the control group,the expression levels of ERα protein in the CGLM low,medium and high dose groups were increased,the difference between the medium and high dose groups was statistically significant（P〈0. 01）. Conclusion CGLM Decoction has a inhibitory effect on the growth of MCF-7 cells and is positively correlated with the dose concentration,and its mechanism may be related to the increase of the content of ERα mRNA and the up-regulated expression of ERα protein.