食管鳞癌中SERPINB5表达降低的作用机制及其临床意义

doi:10.3969/j.issn.1004-616x.2021.02.004

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (2): 101-109.doi: 10.3969/j.issn.1004-616x.2021.02.004

食管鳞癌中SERPINB5表达降低的作用机制及其临床意义

张娜, 范志露, 杨荔艳, 常晨, 蔡岩, 郝佳洁, 王明荣

国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院, 分子肿瘤学国家重点实验室, 北京 100021

收稿日期:2021-02-01 修回日期:2021-02-26 出版日期:2021-03-30 发布日期:2021-04-12
通讯作者: 王明荣,E-mail:wangmr2015@126.com;郝佳洁,E-mail:hjj8173@126.com E-mail:wangmr2015@126.com;hjj8173@126.com
作者简介:张娜,E-mail:13602006330@163.com。
基金资助:
国家重点研发计划项目（2017YFE0112100）；国家自然科学基金（81930077）；中国医学科学院医学与健康科技创新工程（2019-I2M-1-003）

Clinical significance of decreased SERPINB5 expression in esophageal squamous cell carcinomas

ZHANG Na, FAN Zhilu, YANG Liyan, CHANG Chen, CAI Yan, HAO Jiajie, WANG Mingrong

State Key Laboratory of Molecular Oncology, Center for Cancer Precision Medicine, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China

Received:2021-02-01 Revised:2021-02-26 Online:2021-03-30 Published:2021-04-12

摘要/Abstract

摘要： 目的：探索SERPINB5在食管鳞癌中的表达变化、作用机制及其临床意义。方法：利用癌症基因组图谱（TCGA）和基因表达库（GEO）数据库的RNA表达数据、以及临床标本的Western blot分析，研究食管鳞癌组织中SERPINB5的mRNA和蛋白表达情况，并分析其表达水平与临床病理指标的相关性；通过细胞增殖和克隆形成实验，设置亲本组、空载组和SERPINB5过表达组，在体外检测SERPINB5对食管癌细胞增殖的影响；通过裸鼠移植瘤实验，设置空载组和SERPINB5过表达组，在体内检测SERPINB5对食管鳞癌细胞成瘤能力的影响；利用Western blot检测该分子调控的下游通路及关键分子；进一步利用靶向抑制实验，探讨SERPINB5对食管鳞癌治疗的可能意义。结果：与正常食管上皮相比，食管鳞癌组织中SERPINB5的mRNA和蛋白水平均明显降低（P<0.01），且其mRNA低表达与肿瘤低分化（P=0.004）和较晚的分期（P=0.037）显著相关。体外及体内实验结果显示，与对照组比较，SERPINB5过表达可显著降低食管鳞癌细胞的增殖和集落形成、以及裸鼠成瘤能力（P<0.05）。分子水平检测表明，过表达SERPINB5显著降低AKT和mTOR的磷酸化水平。通过靶向抑制实验，观察到SERPINB5过表达可显著增强食管鳞癌细胞对Aurora A抑制剂Alisertib和mTOR抑制剂Everolimus的敏感性（P<0.01）。结论：SERPINB5在食管鳞癌中的表达降低，提高SERPINB5的表达水平，尤其是与Aurora A或mTOR抑制剂的联合使用，可作为食管鳞癌潜在的治疗策略。

关键词: SERPINB5, 食管癌, 细胞增殖, 抑癌基因, 药物敏感性

Abstract: OBJCTIVE: To investigate expression and role of SERPINB5 in esophageal squamous cell carcinomas (ESCC). METHODS: Expression of SERPINB5 mRNA in ESCC tissues was analyzed using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. ESCC specimens and their matched morphologically normal operative margins were used to detect SERPINB5 protein expressions using the Western blot technique. Relationship between SERPINB5 expressions and the clinicopathological parameters of ESCC patients were analyzed. Furthermore,ESCC cells were transfected with an SERPINB5 over-expression vector,the empty vector or without any vectors (parental). These cells were used to detect proliferation and clony formation to determine the roles of SERPINB5. Effects of SERPINB5 on tumorigenesis were detected using the nude mouse xenograft model in vivo. Western blot was used to analyze changes in downstream pathways from over-expression of SERPINB5. Finally,potentials of SERPINB5 in treatment of ESCC through targeted inhibition were investigated. RESULTS: Expression levels of both the SERPINB5 mRNA and protein were down regulated in ESCC compared with the normal tissues (P<0.01). In addition,lower SERPINB5 mRNA levels were correlated with lower degrees of tumor differentiation (P=0.004) but higher clinical stages (P=0.037). From our in vitro and in vivo studies,our results show that over-expression of SERPINB5 significantly inhibited the proliferation and colony formation of ESCC cells,as well as the growth of tumor xenografts (P<0.05). At the molecular level,over-expression of SERPINB5 decreased the phosphorylation levels of AKT and mTOR. Data from targeted inhibition experiments show that the over expression of SERPINB5 significantly increased the sensitivities of ESCC cells to Alisertib (Aurora A inhibitor) or Everolimus (mTOR inhibitor) (P<0.01). CONCLUSION: SERPINB5 acted as a tumor suppressor in ESCC. Elevating the expression of SERPINB5 in tumor cells might be a potential strategy for the treatment of ESCC.

Key words: SERPINB5, esophageal squamous cell carcinoma, cell proliferation, tumor suppressor, drug sensitivity

中图分类号:

R735.1

张娜, 范志露, 杨荔艳, 常晨, 蔡岩, 郝佳洁, 王明荣. 食管鳞癌中SERPINB5表达降低的作用机制及其临床意义[J]. 癌变·畸变·突变, 2021, 33(2): 101-109.

ZHANG Na, FAN Zhilu, YANG Liyan, CHANG Chen, CAI Yan, HAO Jiajie, WANG Mingrong. Clinical significance of decreased SERPINB5 expression in esophageal squamous cell carcinomas[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(2): 101-109.

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食管鳞癌中SERPINB5表达降低的作用机制及其临床意义

Clinical significance of decreased SERPINB5 expression in esophageal squamous cell carcinomas

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