消痰解郁方对乳腺癌前病变MCF-10AT细胞增殖与凋亡及对PTEN蛋白和mRNA表达的影响

Effect of Xiaotanjieyu recipe on the proliferation and apoptosis of MCF-10AT cells and the expression of PTEN protein and mRNA before cancerous lesion in breast cancer

目的 通过研究消痰解郁方对乳腺癌前病变MCF-10AT细胞的生长抑制及诱导凋亡作用和对PTEN蛋白及mRNA表达的影响，探讨消痰解郁方治疗乳腺癌前病变的机制。方法 采用CCK-8法观察24、48 h中药组、西药组、对照组药物分别对乳腺癌前病变MCF-10AT细胞的生长抑制率和凋亡率的影响。Western blot方法检测各组干预后PTEN蛋白表达变化，Q-PCR测定各组PTEN蛋白mRNA的表达情况。结果 （1）中药组、西药组作用于MCF-10AT 24 h、48 h的OD值均低于对照组，差异有统计学意义（P〈0.05），抑制作用呈时间依赖性，且中药组抑制率高于西药组，差异有统计学意义（P〈0.05）。（2）药物干预48 h，细胞凋亡率中药组为（25.40±4.69）%，西药组为（16.93±1.29）%，与对照组（2.63±0.29）%相比明显增高，差异有统计学意义（P〈0.05），中药组比西药组诱导凋亡率高，差异有统计学意义（P〈0.05）；3.中药组与西药组作用于细胞MCF-10AT细胞48 h后，2组PTEN蛋白的表达量明显增加，且中药组的PTEN蛋白表达量增加较西药组明显。PTENmRNA中药组为1.31±0.04，西药组为0.61±0.11均较对照组0.13±0.13明显增加，其中中药组较西药组明显增加，差异有统计学意义（P〈0.05）。结论 消痰解郁方对MCF-10AT细胞有抑制增殖、促进凋亡的作用，其机制可能与上调PTEN蛋白与基因表达水平有关。

Objective To investigate the mechanism of Xiaotanjieyu recipe（XTJYR）in the inhibition of precancerous lesion of breast cancer through the study of its effects on the growth and apoptosis of MCF-10AT cells, as well as its effects on expressions of PTEN protein and mRNA.Methods Changes in the inhibition rate of MCF-10 AT cells growth in precancerous lesion of breast cancer after medication were closely observed at hour 24 and 48, by using CCK-8 method in the 3 groups, i. e. the herbal medicine group （or the XTJYR group）, the western medicine group （or the tamoxifen group） and the control group. Western blot was used to detect changes in the expression of PTEN, and Q-PCR method was employed to detect the expression of PTEN and mRNA in the 3 groups following intervention.Results （1） OD values of the XTJYR group and the tamoxifen group were all lower than those of the control group, with statistical significance （P〈0.05）. The inhibition was time-dependent, furthermore, the inhibition rate of the XTJYR group was higher than that of the tamoxifen group, and statistical significance could be noted, when comparisons were made between them （P〈0.05）. （2） At hour 48 after drug intervention, the rates of apoptosis for the XTJYR group and the tamoxifen group were respectively （25.40±4.69）% and （16.93±1.29）%, which were all significantly higher, as compared with those of the control group （2.63±0.29%）, also with statistical significance （P〈0.05）. The apoptosis rate of the XTJYR group was higher than that of the tamoxifen group and statistical significance could be seen, when comparisons were made between them （P〈0.05）. （3） The expression levels of PTEN were significantly increased in both the XTJYR group and the tamoxifen group 48 hours after intervention on MCF-10AT cells, and the PTEN expression level of the XTJYR group was greater than that of the tamoxifen group. PTEN mRNA expression level of the XTJYR group was （1.31±0.04）, while that of the tamoxifen group was 0.61±0.11, which were all higher than that of the control group （0.13±0.13） , and the expression level of the XTJYR group was higher than that of the tamoxifen group. Statistical significance could be found, when comparisons were made between them （P〈0.05）.Conclusions Xiaotanjieyu recipe proved to have the effects of inhibiting proliferation and promoting apoptosis, the mechanism of which might be related with the up-regulation of PTEN protein and the elevated level of gene expression.