组蛋白去乙酰化酶抑制剂SNDX-275对乳腺癌BT474细胞增殖抑制作用  

The effect of HDAC inhibitor SNDX-275 on inhibiting breast cancer BT474 cell proliferation

作　　者：尹江[1] 刘浩[1] 邓敏[1] 贺智敏[1] 

机构地区：[1]广州医科大学附属肿瘤医院肿瘤研究所,510095

出　　处：《国际肿瘤学杂志》2015年第8期561-565,共5页Journal of International Oncology

基　　金：国家自然科学基金（81472763）;广州市科技计划项目（2014J4100061）

摘　　要：目的明确组蛋白去乙酰化酶抑制剂SNDX-275对ErbB2阳性乳腺癌BT474细胞的增殖抑制作用及其机制。方法以磷酸盐缓冲液（PBS）处理的BT474细胞作为对照组、SNDX-275处理的BT474细胞作为实验组，使用终浓度为0、0.5、1.0、2.0、3.0、4.0 μmol/L的SNDX-275处理细胞，利用MTS实验、克隆形成实验检测细胞增殖能力。采用Western blotting实验检测ErbB2、ErbB3、p-Akt的蛋白表达，实时荧光定量PCR检测miR-125a、miR-125b的表达。MTS实验检测SNDX-275对转染miR-125抑制剂的乳腺癌BT474细胞的增殖抑制作用。结果MTS实验检测结果发现SNDX-275能明显抑制细胞的增殖且具有浓度依赖性，4.0 μmol/L的SNDX-275对细胞的抑制率约（68.00±4.45）%。平板克隆实验结果表明，SNDX-275能明显抑制乳腺癌BT474细胞克隆的形成。Western blotting结果显示SNDX-275明显抑制ErbB2、ErbB3、p-AkT的表达。实时荧光定量PCR分析发现，与PBS处理的BT474细胞对比，2 μmol/LSNDX-275 BT474细胞分别上调miR-125a、miR-125b约3.22±1.17倍、5.42±0.38倍，差异均具有统计学意义（t=4.338,P=0.049；t=21.805,P=0.002）。MTS实验结果显示，与PBS对照组相比，SNDX-275组和转染miR-125抑制剂组对乳腺癌细胞BT474的抑制率分别为（56.97±3.56）%、（10.67±2.21）%，两组之间差异具有统计学意义（t=-10.993，P=0.008）。结论SNDX-275通过上调miR-125a、miR-125b抑制ErbB2-ErbB3-Akt信号通路，进而抑制ErbB2阳性乳腺癌BT474细胞的增殖。Objective To explore the effect and molecular mechanism of HDAC inhibitor SNDX-275 inhibiting cell proliferation in ErbB2-overexpressing breast cancer BT474 cells. Methods Breast cancer BT474 cells were treated with HDAC inhibitor SNDX-275, setting as test group, and the ceil line treated with phosphate buffered saline （PBS） as control. The concentration of SNDX-275 were 0, 0. 5, 1.0, 2.0, 3.0, 4.0 μmol/L respectively. Cell proliferation was analyzed by MTS assay and colony formation assay, the expressions of ErbB2, ErbB3, p-Akt were analyzed by Western blotting, and the expressions of miR-125a, miR-125b were analyzed by RT-PCR. After transfecting miRNA125 inhibitor into BT474 cells, the inhibition rate of SNDX-275 was tested by MTS assay. Results MTS result showed that SNDX-275 inhibited cell proliferation in BT474 cells in a dose-dependent manner. The inhibition rate of 4.0 Ixmol/L SNDX-275 was about （68.00 ± 4.45 ） %. Clone assay indicated SNDX-275 could inhibit the proliferation of BT474 cells. Western blotting result indicated that SNDX- 275 significantly inhibited the protein expressions of ErbB2, ErbB3 and p-Akt, RT-PCR result illustrated 2 μmol/L SNDX-275 could increase the expressions of miR-125a and miR-125b about 3.22 ± 1.17, 5.42 ±0.38 times compared with the PBS control respectively, the difference has a statistical significance （t = 4. 338, P = 0. 049 ;t = 21. 805 ,P = 0. 002）. MTS result indicated that compared with the PBS control, the inhibition rate of SNDX-275 group was （56.97 ± 3.56）%, while the inhibition rate of SNDX-275 and miRNA125 inhibitor groupwas （ 10.67 ± 2.21 ） %, with a statistical significance（ t = - 10.993 ,P = 0. 008）. Conclusion SNDX-275 could inhibit cell proliferation of ErbB2-overexpressing breast cancer BT474 cells, by inhibiting ErbB2-ErbB3-Akt signal pathway through up-regulating miR-125a and miR-125b.

关 键 词：乳腺肿瘤 细胞增殖 微RNAS 

分 类 号：R737.9[医药卫生—肿瘤]