不同雌激素受体表达状态的子宫内膜癌细胞中转录因子的活性分析

Analysis different transcriptional factors in different phenotype endometrial cancer cells

目的分析并比较不同雌激素受体（ER）表达状态的子宫内膜癌细胞中转录因子的活性。方法采用实时荧光定量lit-PCR技术检测子宫内膜癌细胞系RL-952[ERα、ERβ表达均阳性（±）]、HEC-1A[ERa表达弱阳性（±）、ERβ表达阴性（-）]、HEC-1B[ERα、ERβ表达均（-）]细胞中ERαmRNA的表达。采用345通量转录因子芯片检测RL-952、HEC-1A、HEC-1B细胞中转录因子的活性，采用酶联免疫吸附试验（ELISA）方法检测不同转录活性的转录因子NFKBp65、p38MAPK以验证芯片检测结果。结果ERαmRNA在RL-952、HEC-1A、HEC-1B细胞中表达水平依次递减，分别为（6780±282）、（684±84）、（168±38）copy／ng。转录因子NFκBp65、p38MAPK的转录活性采用ELISA方法检测（分别为2．0±0．4、0．9±0．5，P=0．020）与芯片检测（分别为3003±530、882±538，P=0．017）结果一致。在345个转录因子中，筛选出与ER功能相关的差异表达的转录因子共28个，与ER（±）的RL-952细胞相比，ER（-）的HEC-1A、HEC-1B细胞中转录因子活性同时上调的有13种，同时下调的有15种。转录因子TTF（1）-1、NRF-1、TCE的活性与ERamRNA表达水平呈明显线性正相关关系（r=0．523，P=0．037），而转录因子RFX123和Ikaros的活性与ERamRNA表达水平呈明显非线性负相关关系（r=-0．312，P=0．041）。结论转录因子芯片检测是筛选子宫内膜癌致病机制中主要转录因子的先进技术。转录因子TTF（1）-1、NRF-1、TCE可能与ER（±）子宫内膜癌中的信号传导通路相关；转录因子RFX123和Ikaros可能与ER（-）子宫内膜癌中的信号传导通路相关。

Objective To analysis the activity of transcriptional factors in endometrial cancer cell lines with different estrogen receptor subtypes. Methods The mRNA levels of estrogen receptor （ER）was detected by quantitative RT-PCR , and the activity of transcriptional factors was also analysed by 345-channel protein/DNA array in RL-952 （the expression status of ERα and ERβ both positive）, HEC-1A [ ERα（ ± ）, while ERβ negative ] and HEC-1 B （ ERα and ERβ both negative）. The transcription factors of NFκBp65 and p38MAPK with different activity were tested by enzyme-linked immunosorbent assay（ELISA） to confirm the results of protein/DNA array. Results The mRNA levels of ERa in RL-952, HEC-1A and HEC-1B were （6780 ± 282 ）, （ 684 ± 84 ） and （ 168 ± 38 ） copy/ng, respectively. Among 345 candidate transcriptional factors, there were 28 factors associated with ER status. Compared with RL-952 cells, 13 transcriptional activity factors were concomitantly up-regulation, while 15 concomitantly down-regulation in HEC-1A and HEC-1B cells. Transcriptional activities of TTF （ 1 ） -1, NRF-1, TCE were significantly correlated with the high-expression status of ERa mRNA（ r = 0. 523, P = 0. 037 ）,while RFX123 and Ikaros were significantly correlated with the low-expression status of ERα mRNA （ r = - 0. 312, P = 0. 041 ）. Conclusion Transcriptional factors of TFF（ 1 ）-1, NRF-1, TCE may be associated with ER-mediated signal pathway, while RFX123 and Ikaros may be associated with non ER-mediated signal pathway in endometrial cancer.