实用医学杂志 ›› 2022, Vol. 38 ›› Issue (17): 2138-2144.doi: 10.3969/j.issn.1006⁃5725.2022.17.005

• 基础研究 • 上一篇    下一篇

E2F转录因子7在甲状腺乳生物学作用头状癌中的表达及

范会利1 赵如华1 张赫1 林旭1 薛刚2 吴靖芳1    

  1. 河北北方学院1 形态学实验室,2 耳鼻咽喉头颈外科教研室(河北张家口075000)

  • 出版日期:2022-09-10 发布日期:2022-09-10
  • 通讯作者: 吴靖芳 E⁃mail:wjfxg@163.com
  • 基金资助:
    河北省自然科学基金(编号:H2018405054);河北北方学院校级项目(编号:YB020019)

Expression and biological role of E2F7 in papillary thyroid carcinoma

FAN Huili*,ZHAO Ruhua,ZHANG He, LIN Xu,XUE Gang,WU Jingfang.   

  1. Laboratory of Morphology,Hebei North University,Zhangjiakou 075000,China

  • Online:2022-09-10 Published:2022-09-10
  • Contact: WU Jingfang E⁃mail:wjfxg@163.com

摘要:

目的 探讨 E2F 转录因子 7(E2F7)在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)中的 作用及分子机制。方法 生物信息学分析 E2F7 PTC 的关系;免疫组织化学方法检测 PTC 及癌旁组织 E2F7 的表达;Western blot(WB)检测人甲状腺癌细胞系 K1、TPC⁃1 BCPAP 细胞以及正常人甲状腺 Nthy⁃ ori 3⁃1 细胞系 E2F7 表达水平;以生长曲线、克隆形成实验和划痕实验检测沉默 E2F7 K1 细胞增殖和迁 移能力的影响;流式细胞术检测沉默 E2F7 K1 细胞周期和凋亡的影响;qRT⁃PCR 检测其对周期蛋白 cyclin D1、cyclin E1、EIF4A3 及周期蛋白依赖性激酶 4(CDK4)基因转录的影响,以 WB 检测 E2F7 对周期蛋 白和通路蛋白 Akt、pAkt 的影响。结果 E2F7 PTC 组织中表达水平较癌旁组织高表达,其中临床分期及 淋巴结转移与 E2F7 表达水平有统计学意义(P < 0.05);K1 细胞 E2F7 的表达明显高于正常甲状腺细胞; E2F7 沉默组 K1细胞蛋白表达水平相比对照组明显降低(P < 0.01);增殖、迁移能力较对照组受到抑制(P < 0.05);E2F7 沉默组周期蛋白转录和翻译水平较对照组降低;通路关键蛋白 Akt pAkt 表达水平较对照组 下调;E2F7 沉默组细胞周期较对照组细胞周期进展出现阻滞,凋亡率升高。结论 E2F7 高表达与 PTC 生、发展有关;E2F7基因可能通过Akt 信号通路促进K1细胞增殖、抑制凋亡。

关键词:

甲状腺乳头状癌, E2F转录因子7, 细胞周期, 细胞凋亡, Akt 信号通路

Abstract:

Objective To investigate the role and molecular mechanism of E2F7 in papillary thyroid carcinoma(PTC). Methods The correlation between E2F7 and TC was predicted by bioinformatics analysis. The expression of E2F7 in PTC and adjacent tissues was detected by immunohistochemistry. Western blotting(WB was used to detect the expression level of E2F7 in human PTC K1,TPC⁃1 and BCPAP cell lines and in normal human thyroid Nthy⁃ori3⁃1 cell line. The effect of silencing E2F7 on the proliferation and migration of K1 cells was detected by growth curve,clone formation assay and scratch assay. Flow cytometry was used to detect the effect of silencing E2F7 on K1 cell cycle and apoptosis. The transcription of cyclin cyclinD1,cyclinE1,EIF4A3 and cyclin⁃dependent kinase 4(CDK4)genes was detected by qRT⁃PCR,and the effect of E2F7 on cyclin and pathway proteins Akt,pAkt by WB. Results The expression level of E2F7 in PTC tissues is higher than that in adjacent tissues,clinical stage and lymph node metastasis were statistically significant with E2F7 expression level;The expression of E2F7 in K1 cells is significantly higher than that in normal thyroid cells;the protein expression level of K1 cells in the E2F7 silence group is compared with that in the control group compared with the control group The proliferation and migration abilities were inhibited in the E2F7 silence group;The transcription and translation levels of cyclins in the E2F7 silence group were lower than the control group;The expression levels of key pathway proteins Akt and pAkt were down ⁃ regulated compared with the control group;The cycle progression was blocked and the rate of apoptosis increased. Conclusions The high expression of E2F7 in PTC tissue is related to the occurrence and development of PTC;E2F7 silencing causes K1 cell cycle arrest,promotes apoptosis and inhibits proliferation through Akt signaling pathway.

Key words:

papillary thyroid carcinoma, E2F transcription factor 7, cell cycle, apoptosis, Akt signaling pathway