中国癌症杂志 ›› 2019, Vol. 29 ›› Issue (4): 265-271.doi: 10.19401/j.cnki.1007-3639.2019.04.004

• 论著 • 上一篇    下一篇

TBRG4在肺腺癌中的作用及其机制研究

叶 柯,郑晓丽,葛 红,李 雪,王 浩   

  1. 郑州大学附属肿瘤医院放疗科,河南 郑州 450000
  • 出版日期:2019-04-30 发布日期:2019-05-17
  • 通信作者: 葛 红 E-mail: HZHSKY66@163.com
  • 基金资助:
    国家自然科学基金(81773230);国家自然科学基金(81372436)。

The role of TBRG4 in lung adenocarcinoma and its mechanism

YE Ke, ZHENG Xiaoli, GE Hong, LI Xue, WANG Hao   

  1. Department of Radiation Oncology, Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou 450000, Henan Province, China
  • Online:2019-04-30 Published:2019-05-17
  • Contact: GE Hong E-mail: HZHSKY66@163.com

摘要: 背景与目的:转化生长因子β调节子4(transforming growth factor β regulator 4,TBRG4)是编码转化生长因子β(transforming growth factor β,TGF-β)的调节子;近年来研究发现,TBRG4表达异常与肿瘤密切相关,但有关TBRG4与肺腺癌关系的研究较少,因此该研究旨在探讨TBRG4在肺腺癌中的作用及其相关机制。方法:收集郑州大学附属肿瘤医院非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的肿瘤组织和癌旁组织样本,采用免疫组织化学方法检测TBRG4的表达。以慢病毒为载体构建TBRG4基因特异性干扰载体,并将特异性干扰载体及空载体转染NCI-H1299细胞,采用细胞计数试剂盒(cell counting kit-8,CCK-8)检测各组细胞生长情况;采用细胞克隆形成实验比较各组细胞克隆形成能力;采用Annexin Ⅴ-FITC/PI双染法结合流式细胞术检测细胞凋亡;PI/RNase单染检测细胞周期;采用蛋白质印迹法(Western blot)检测10号染色体上缺失的磷酸酶及张力蛋白同源物(phosphatase and tensin homolog deleted on chromosome ten,PTEN)信号通路相关蛋白表达情况。结果:TBRG4蛋白在NSCLC患者癌组织中表达升高;TBRG4敲低后H1299细胞抑制了H1299细胞的增殖及细胞克隆形成能力,并促进细胞凋亡,同时导致H1299细胞周期停滞于G0/G1期;PTEN、RAP1A及IGF2蛋白表达上调,差异有统计学意义(P<0.05)。结论:TBRG4可能参与肺癌的发生、发展过程,其在NSCLC患者肺癌组织中的表达显著高于癌旁组织。TBRG4与肺癌细胞的增殖、调亡等生物学行为密切相关,可能成为肺癌治疗的潜在靶点。

关键词: 转化生长因子&beta, 调节子4, 非小细胞肺癌, RNA干扰, 细胞增殖, 细胞凋亡

Abstract: Background and purpose: Transforming growth factor β regulator 4 (TBRG4) is a regulator of transforming growth factor β. In recent years, studies have found that TBRG4 expression is closely related to tumors, but there are few studies on the relationship between TBRG4 and lung adenocarcinoma. Therefore, this paper aimed to study the role of TBRG4 in lung adenocarcinoma and related mechanism. Methods: Clinical samples of patients with non-small cell lung cancer (NSCLC), including tumors and adjacent tissues, were collected. Immunohistochemistry was used to detect the expression of TBRG4. The TBRG4 genespecific interference vector was constructed with lentivirus as vector, and the specific interference vector and empty vector were transfected into NCI-H1299 cells. Cell counting kit-8 (CCK-8) was used to detect the cell growth in each group. The cell clone formation experiment was used to compare the cell clone formation ability in each group. Apoptosis was detected by Annexin Ⅴ-FITC/PI double staining combined with flow cytometry. PI/RNase single staining was used to detect cell cycle. Western blot was used to detect phosphatase and tensin homolog deleted on chromosome ten (PTEN) signaling pathway-related protein expression. Results: TBRG4 protein was up-regulated in NSCLC patients' cancer tissues. Downregulation of TBRG4 inhibited H1299 cell proliferation and cell clonality, promoted apoptosis, and caused H1299 cell cycle arrest in G0/G1 phase. The protein expression levels of PTEN, RAP1A and IGF2 were up-regulated (P<0.05). Conclusion: TBRG4 may be involved in the development of lung cancer, and its expression in lung cancer tissues of patients with NSCLC is significantly higher than that in adjacent tissues. TBRG4 plays an important role in the biological behavior, such as proliferation and apoptosis of lung cancer cells. TBRG4 may be a  potential target for the treatment of lung cancer.

Key words: TBRG4, Non-small cell lung cancer, RNA interference, Cell proliferation, Cell apoptosis