| 沈方倩,隋晓馨,何乐伟,曹 莹,席晓薇.上皮性卵巢癌中TFAP2A对hTERT表达的调节及其机制研究[J].,2020,(22):4201-4206 |
| 上皮性卵巢癌中TFAP2A对hTERT表达的调节及其机制研究 |
| TFAP2A Might Active hTERT in EOC Cells without through PI3K/AKT Signaling Pathway in Epithelial Ovarian Cancer |
| 投稿时间：2020-05-23 修订日期：2020-06-17 |
| DOI：10.13241/j.cnki.pmb.2020.22.001 |
| 中文关键词: TFAP2A hTERT PI3K/AKT 上皮性卵巢癌 |
| 英文关键词: TFAP2A hTERT PI3K/AKT Epithelial ovarian cancer |
| 基金项目:上海市科委重点研究项目（No.2013ZYJB9201） |
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| 中文摘要: |
| 摘要 目的：探讨在上皮性卵巢癌中TFA2A对hTERT表达的调节和作用机制。方法：采用免疫组织化学方法检测TFAP2A和hTERT蛋白在卵巢正常、交界及上皮性卵巢癌组织中的表达，采用Western Blot和qRT-PCR技术检测hTERT在敲减TFAP2A基因的SKOV3、CAOV3细胞中的表达水平、检测hTERT在过表达TFAP2A基因的HO8910细胞中的表达水平。在干扰TFAP2A的CAOV3细胞中或过表达TFAP2A的HO8910细胞中分别加入PI3K/AKT信号通路激动剂740-YP或抑制剂LY294002，检测相关蛋白表达变化，探讨TFAP2A、hTERT与PI3K/AKT信号通路的关系。结果：TFAP2A在71.88%的上皮性卵巢癌组织中呈高表达，hTERT在78.12%的上皮性卵巢癌组织中呈高表达； 将hTERT 和TFAP2A的免疫组化评分行Pearson相关性分析，两者间相关系数r=0.78，P＜0.001。Western Blot和qRT-PCR的结果均显示，在SKOV3和CAOV3卵巢癌细胞中，敲减TFAP2A后，hTERT的表达均明显下降，而在HO8910卵巢癌细胞中，增强TFAP2A基因表达后，hTERT的表达均明显上升。在CAOV3和HO8910处理细胞中，分别使用PI3K/AKT信号通路激动剂740-YP 或阻滞剂LY294002处理后，Western Blot 检验hTERT和PI3K/AKT通路蛋白的表达，发现激动剂740-YP 或阻滞剂LY294002可以逆转敲减或过表达TFAP2A引发的PI3K/AKT通路蛋白表达下调或上调，但不能逆转hTERT蛋白表达下调或上调。结论：在卵巢肿瘤组织中，TFAP2A和hTERT在上皮性卵巢癌组织中均呈高表达，且hTERT的表达和TFAP2A成正相关，在上皮性卵巢癌细胞中TFAP2A可调节hTERT的表达，且TFAP2A对hTERT的表达的调节不经由PI3K/AKT通路。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the regulation and mechanism of TFA2A on hTERT expression in epithelial ovarian cancer. Methods: We detected the expression level of TFAP2A and hTERT in normal ovarian epithelial tissues, borderline ovarian tumor tissues as well as epithelial ovarian cancer tissue by an IHC assay. The expression of TFAP2A gene in CAOV3 and SKOV3 cell lines were silenced by using small interfering RNA technique.The expression of TFAP2A in HO8910 cell line were overexpressed by overexpression plasmid. Western Blot and qRT-PCR were used to test the expression of hTERT in TFAP2A-silenced SKOV3 and CAOV3 cells or TFAP2A-overexpressed HO8910 cells. The relationship between TFAP2A, hTERT and PI3K/AKT signaling pathways was explored by using the agonist 740-YP and inhibitor LY294002 of the PI3K/AKT signalling pathway in TFAP2A-silenced CAOV3 cells or in TFAP2A-overexpressed HO8910. Results: By immunohistochemistry assays, we found that TFAP2A was highly expressed in 71.88% epithelial ovarian cancer tissues, as well as hTERT was highly expressed in 78.12% of epithelial ovarian cancer tissues. By Pearson correlation analysis, We also found that the expression levels of hTERT were consistent with TFAP2A in different ovarian tissues, and the correlation coefficient was 0.78. Western Blot and qRT-PCR showed that the expression levels of hTERT was significantly decreased in TFAP2A-silenced CAOV3 and SKOV3 cells, while it was increased in TFAP2A-overexpressed HO8910. Western Blot showed that the hTERT would not significantly increased or decreased as the PI3K/AkT proteins after using PI3K/AkT signaling pathway agonist 740-YP or blocker LY294002. Conclusion: TFAP2A up-regulates hTERT expression in epithelial ovarian cancer cells do not by way of PI3K/AKT signaling. |
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