上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (10): 1134-.doi: 10.3969/j.issn.1674-8115.2019.10.006

• 论著·基础研究 • 上一篇    下一篇

谷丙转氨酶2对胃癌顺铂治疗抵抗的影响及其作用机制

袁 园，糜 军   

  1. 上海交通大学基础医学院生物化学与分子细胞生物学系，上海 200025
  • 出版日期:2019-10-28 发布日期:2019-11-22
  • 通讯作者: 糜 军，电子信箱：jmei@sjtu.edu.cn。
  • 作者简介:袁 园（1992—），女，硕士生；电子信箱：yuanyuan201609@126.com。
  • 基金资助:
    国家自然科学基金面上项目（81372194）

Effects and mechanisms of glutamic-pyruvic transaminase 2 on cisplatin resistance in gastric cancer

YUAN Yuan, MEI Jun   

  1. Department of Biochemistry and Molecular Cell Biology, Shanghai Jiao Tong University College of Basic Medical Science, Shanghai 200025, China
  • Online:2019-10-28 Published:2019-11-22
  • Supported by:
    General Program of National Natural Science Foundation of China, 81372194

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摘要： 目的·研究谷丙转氨酶2（glutamic-pyruvic transaminase 2，GPT2）在胃癌顺铂治疗抵抗中的作用及其机制。方法·利用Kaplan Meier-Plotter数据库分析GPT2的表达与胃癌不良预后之间的关系，采用实时荧光定量PCR（quantitative real-time PCR，qPCR）、蛋白质印迹（Western blotting）和免疫组织化学（immunohistochemistry，IHC）技术检测GPT2在胃癌细胞和临床胃癌组织样本中的表达情况。采用CCK-8试剂盒检测不同浓度的顺铂对人胃癌细胞和正常胃黏膜细胞的细胞毒性。分别在顺铂敏感的MKN28细胞和不敏感的MKN45细胞中构建GPT2过表达和低表达细胞系，并采用CCK-8试剂盒、细胞克隆形成实验、Western blotting检测顺铂对GPT2过表达和低表达胃癌细胞的细胞毒性、肿瘤细胞的干性及干性相关信号通路上关键蛋白的变化。结果·GPT2的高表达与胃癌预后呈负相关。GPT2表达高的细胞株对顺铂治疗不敏感，GPT2表达低的细胞株则相反。GPT2的过表达可降低顺铂治疗的敏感性，而低表达GPT2则相反。过表达GPT2可激活细胞外调节蛋白激酶（extracellular regulated protein kinases，ERK）信号通路、上调干性相关分子标志物SRY盒2（SRY-box 2，SOX2）和Nanog同源盒（Nanog homeobox，NANOG）的表达，同时可增强细胞的克隆形成能力，而低表达GPT2则反之。结论·GPT2的表达与顺铂治疗敏感性有关。过表达GPT2可通过激活ERK信号通路，上调干性相关分子标志物SOX2和NANOG的表达，实现胃癌细胞对顺铂治疗的抵抗。

关键词: 谷丙转氨酶2, 胃癌, 顺铂, 治疗抵抗

Abstract:

Objective · To investigate the role and mechanism of glutamic-pyruvic transaminase 2 (GPT2) on cisplatin resistance in gastric cancer. Methods · The Kaplan Meier-Plotter database was used to analyze the relationship between GPT2 and poor prognosis of gastric cancer. The s of GPT2 in gastric cancer cells and tissues were detectedquantitative real-time PCR (qPCR), Western blotting and immunohistochemistry (IHC). The cytotoxicity of cisplatin at different concentrations to human gastric cancer cells and normal gastric epithelial cells was detectedCCK-8 assay. GPT2 over and knockdown cell lines were constructed in cisplatin sensitive MKN28 cells and insensitive MKN45 cells, respectively. CCK-8 assay, colony formation assay and Western blotting were performed to evaluate the cellular cytotoxicity, stemness of cancer cells and the changes of key proteins in stemness-related signaling pathways in GPT2 over and knockdown cell lines, respectively. Results · The high of GPT2 was negatively correlated with the survival of gastric cancer patients. Gastric cancer cells with high of GPT2 were resistant to cisplatin, while cells with low of GPT2 were sensitive to cisplatin. Over of GPT2 could decrease the cell sensitivity to cisplatin, nevertheless knockdown of GPT2 could increase the cell sensitivity to cisplatin. Meanwhile, the further study revealed that over of GPT2 could activate the extracellular regulated protein kinases (ERK) signaling pathway, up-regulate the of SRY-box 2 (SOX2) and Nanog homeobox (NANOG), and enhance the ability of colony formation, while knockdown of GPT2 could inhibit ERK signaling pathway, reduce the of SOX2 and NANOG, and suppress the ability of colony formation. Conclusion · GPT2 are related to the sensitivity of cisplatin treatment. Over of GPT2 can increase the resistance of gastric cancer to cisplatin treatmentactivating ERK signaling pathway and up-regulating the of SOX2 and NANOG.

Key words: glutamic-pyruvic transaminase 2 (GPT2), gastric cancer, cisplatin, treatment resistance

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