雌激素对人乳腺癌细胞株MCF-7 KLK 6 mRNA和蛋白表达水平的影响

 目的： 探讨雌激素与其拮抗剂三苯氧胺对雌激素受体阳性人乳腺癌细胞株MCF-7组织激肽释放酶6（KLK 6）mRNA和蛋白（hK6）表达水平的影响。方法: 取生长良好的MCF-7细胞，在不同浓度的雌激素（17-βE2）和三苯氧胺存在下培养72 h后，收集细胞，分别采用荧光定量逆转录-聚合酶链反应和流式细胞术，检测KLK6 mRNA和蛋白表达水平的变化。结果: 实时荧光定量RT-PCR表明，当17-βE2浓度在10-12 mol/L时，与乙醇对照组无明显差异(P>0.05), 17-βE2浓度在10^-10 mol/L和10-8 mol/L时，KLK6 mRNA的相对含量显著少于乙醇对照组；流式细胞术分析表明, 代表hK6含量的相对荧光强度（AFI）显著低于乙醇对照组（P<0.01）。三苯氧胺（10^-6 mol/L）作用组KLK6 mRNA 的相对含量显著高于对照组（P<0.01）；hK6含量与对照组相比差异显著（P<0.01）。结论: 雌激素对MCF-7 KLK6 mRNA和蛋白表达水平有一定的下调作用，而三苯氧胺作用相反。

Regulation of estrogen on the expression of KLK6 mRNA and protein in human breast cancer cell line MCF-7

AIM:To investigate the effects of estrogen and tamoxifen on the expression of KLK6 mRNA and protein (hK6) in human breast cancer cell line MCF-7. METHODS:MCF-7 cells were incubated with 17-βE2 and tamoxifen at different concentrations for 72 hours, respectively. The expression levels of kallikrein 6 (KLK6) mRNA and protein were evaluated by fluorescence quantitative RT-PCR and flow cytometry, respectively. RESULTS:Compared with ethanol control, KLK6 mRNA expression levels were significantly decreased when 17-βE2 was added at concentrations of 10^-10 and 10^-8 mol/L (P<0.01). No statistical change was observed when 17-βE2 was at 10^-12 mol/L (P>0.05). Flow cytometry showed the same results. The average fluorescence intensity (AFI) that represents the level of hK6 was decreased after incubated with 17-βE2 (P<0.01). After incubation with tamoxifen, the levels of KLK6 mRNA and hK6 were increased (P<0.01). CONCLUSION:Estrogen down-regulates the expression levels of KLK6 mRNA and protein (hK6), while tamoxifen has an opposite effect.