摘要
目的探讨甲状腺激素受体βΔ(thyroid hormone receptorβΔ,TRβΔ)对大鼠乳腺癌SHZ-88细胞增殖、凋亡的影响及其作用机制。方法将真核表达载体pcDNA3.1-TrβΔ及空载体pcDNA3.1分别转染体外培养的SHZ-88细胞,并给予10 nmol/L T3干预。CCK-8(Cell Counting Kit-8)法分析细胞的增殖活力;Annexin V/PI和JC-1荧光染色,流式细胞仪检测细胞凋亡及细胞线粒体膜电位变化;ELISA法检测细胞组蛋白/DNA碎片;定量PCR检测细胞半胱氨酸天冬氨酸特异性蛋白酶-9(Cysteinyl aspartate specific proteinase-9,Caspase-9)、Caspase-3基因表达;比色法检测Caspase-9和Caspase-3活性。结果与空载体pcDNA3.1转染组相比,pcDNA3.1-TrβΔ转染组明显抑制SHZ-88细胞的增殖、增加细胞组蛋白/DNA碎片和凋亡细胞百分比、降低线粒体膜电位、上调Caspase-9、Caspase-3基因的表达并促进Caspase-9及Caspase-3的活化(P<0.05);给予T3后可明显增强上述作用(P<0.05)。结论TRβΔ可抑制SHZ-88细胞的增殖并促进其发生凋亡,该作用可能通过降低线粒体膜电位,激活Caspase-9及Caspase-3蛋白,启动内源性细胞凋亡途径实现,且该作用受T3调节。
Objective To investigate the effect of thyroid hormone receptor β△（TRβ△） on the proliferation and apoptosis of rat breast cancer cell line SHZ-88 in vitro and related mechanism. Methods The expression plasmid pcDNA3.1-Trβ△ and empty vector pcDNA3.1 were compared in the presence or absence of 10 nmol/L T3. Cell Counting Kit-8 was used to measure SHZ-88 cells activity. The apoptotic rate and mitochondrial transmembrane potential were analyzed by flow cytometry. Histone/DNA fragment was assayed by ELISA. The levels of Cysteinyl aspartate specific proteinase-9（Caspase-9）, Caspase-3 mRNA were detected by quantitative RT-qPCR. The activity of Caspase-9 and caspase-3 were measured using colorimetry. Results Compared with empty vector pcDNA3.1 transfection group, pcDNA3.1-Trβ△ transfection group significantly inhibited the cell proliferation, increased the level of histone/DNA fragment and the percentage of apoptotic cells, up-regulated Caspase-9, Caspase-3 expression and activity, and reduced the mitochondrial transmembrane potential in SHZ-88 cells（P〈0.05）. These effects could be significantly strengthened by T3（P〈0.05）. Conclusion TRβ△ could inhibit the proliferation of SHZ-88 cells and enhance cell apoptosis, which might be via reducing the mitochondrial transmembrane potential, activating Caspase-9 and Caspase-3 protein, and starting endogenous apoptotic pathway, moreover, these effect could be regulated by T3.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2016年第12期1018-1022,共5页
Cancer Research on Prevention and Treatment
基金
国家自然科学基金(81301737)
山东省自然科学基金(ZR2015HL025)
关键词
甲状腺激素受体
TRβΔ亚型
乳腺癌
增殖
凋亡
Thyroid hormone receptor
TRβ△ isoform
Breast cancer
Proliferation
Apoptosis
