组蛋白去乙酰化酶抑制剂MS-275对乳腺癌细胞中ERα表达的影响

The effect of HDAC inhibitor MS-275 on ERα expression of breast cancer cells

目的观察组蛋白去乙酰化酶抑制剂(MS-275)对乳腺癌细胞株MDA-MB-231和MCF-7中ERαmRNA表达水平的影响以及对乳腺癌细胞株增殖、凋亡、侵袭能力的影响。方法以MDA-MB-231和MCF-7乳腺癌细胞株为研究对象,设定非药物处理组和MS-275不同的浓度梯度作用于细胞,进行MTT试验,并得出对癌细胞的生长抑制曲线,然后以1.0μmol/L的MS-275作用于细胞48 h,采用半定量逆转录-聚合酶链反应(RT-PCR)检测用药前后乳腺癌细胞中ERα表达水平的变化,并采用流式细胞仪研究细胞生物学行为的变化。结果 MTT检测结果显示,MS-275对细胞的抑制率随浓度和作用时间的增加而呈明显上升趋势,与阴性对照组相比有显著性差异(P>0.05)。选择用1.0μmol/L浓度MS-275作用48 h后,MDA-MB-231恢复表达ERα,未用药物干预之前不表达ER-α,药物处理后ER-α出现表达,其表达的平均水平为0.32±0.11,P<0.05。MCF-7未经药物干预时表达ERα平均表达水平0.52±0.44,药物处理后平均表达水平0.76±0.53,较未经药物处理的细胞明显升高(P<0.05)。流式细胞检测显示MS-275影响细胞的增殖:可以使乳腺癌细胞的增殖停滞在G1/S期和G2/M期,和阴性对照组及DMSO(MS-275的溶媒)组,细胞凋亡明显,差异有显著性(P<0.05)。结论组蛋白去乙酰化酶抑制剂MS-275通过诱导细胞凋亡而非直接的细胞毒作用产生抗肿瘤活性,同时可以诱导雌激素受体阴性的乳腺癌细胞株恢复部分表达ERα。

Objective To investigate the effects of MS-275,a sort of histone deacetylation inhibitor,acting on breast cancer cells MDA-MB-231 and MCF-7 by making ERα expression level increase or re-express and the effects of MS-275 on proliferation and apoptosis of breast cancer cells.Methods Treat breast cancer cells with MS-275 at different concentration（0.25 μmol/L,0.5 μmol/L,1.0 μmol/L,2.5 μmol/L）,at the same time set the control group to compare.Draw the cells growing inhibiting curves by MTT test after the medicine intervention.Measure the ERα mRNA level by RT-PCR in breast cancer cells before or after the treatment（1.0μmol/L for 48 h）.Then observe the biological behavior of breast cancer cells using Flow Cytometry method.Results MS-275 as a histone de-acetylation inhibitor could inhibit the growth of breast cancer cells significantly and its inhibiting effect contributed to the concentration and function time proved by MTT test.MDA-MB-231,ER-negative breast cancer cell,re-express ERα mRNA after treated with 1.0 μmol/L MS-275 for 48 h.MCF-7,ER-positive breast cancer cells,got an increased expression after the same treatment.And they all had significant differences.Tested by Flow Cytometry method,MS-275 can suspended the breast cancer cells generation at G1/S and G2/M period.This HDAC inhibitor also induce breast cancer cells apoptosis viewed by electron microscope.Conclusion MS-275 is a histone de-acetylation inhibitor.Its anti-tumor effect mostly depended on apoptosis.Meantime,it made a new therapy direction for it can cause ER-negative breast cancer cells re-express ERα.