甲状腺激素受体β△对大鼠肝癌细胞RH-35凋亡和增殖的影响

Effects of thyroid hormone receptor β△ on apoptosis and proliferation of hepatoma RH-35 cells

目的探讨甲状腺激素受体β△（thyroid hormone receptorβ△，TRl3A）对大鼠肝癌细胞系RH-35凋亡和增殖的影响及其作用机制。方法将空载体pcDNA3．1及pcDNA3．1-TRβ△分别转染体外培养的大鼠肝癌细胞系RH-35，并给予10nmol／LT，干预。AnnexinV／PI荧光染色，流式细胞仪检测细胞凋亡率；MTr法测定细胞增殖；实时PCR检测β-连环素编码基因（cateninβ-1，CTNNBl）、衰老标记蛋白．30基因（senescence marker protein-30，SMP-30）、BAK基因（BCL2-antagonist／killer，BAK）的表达。结果T1存在时。TRl3A过表达抑制RH-35细胞的增殖；促进细胞的凋亡；下调CTNNBl和SMP-30基因表达；上调BAK基因表达（P〈0．05）。结论TRl3A可抑制RH-35细胞的增殖并促进其发生凋亡，其机制可能与上调BAK基因，下调CTNNBl和SMP-30基因表达有关，且该作用受T3调节。

Objective To study the effects of TRI3A on apoptosis and proliferation of liver cancer cell line RH-35 from rat in vitro. Methods RH-35 cells were transfected by empty vector peDNA3.1 and expression plasmid pcDNA3.1-TRβ△, then exposure to 10 nmol/L T3. RH-35 cells apoptosis and proliferation were observed by flow eytometry and MTT colorimetric assay; Levels of catenin β-1 （ CTNNB1 ） , senescence marker protein-30 （ SMP-30 ） and BCL2-antagonist/killer （BAK） mRNA evaluation were detected by quantitative real-time RT-PCR （RT-qPCR）. Results In the presence of T3, overexpression of TRβ△ significantly inhibited the proliferation, increased the percentage of apoptotic, down-regulated CTNNB1 and SMP-30 expression, up-regulated BAK expression in RH-35 cells （ P〈 0.05 ）. Conclusion TRβ△ could inhibit the proliferation of RH-35 cells and promote their apoptosis, which may be related to upregulation of BAK genes expression and downregulation of CTNNB1 and SMP-30 gene expression, and these effects could be regulated by Ts.