抑制USP9x表达增强食管癌Ec9706-R细胞的放射敏感性

金迎迎; 王敏聪; 惠文涛; 潘继元; 马红兵

doi:10.19401/j.cnki.1007-3639.2019.07.002

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抑制USP9x表达增强食管癌Ec9706-R细胞的放射敏感性

论著 | 更新时间：2026-01-27

- 抑制USP9x表达增强食管癌Ec9706-R细胞的放射敏感性
- Inhibition of USP9x expression enhances the radiosensitivity of esophageal cancer Ec9706-R cells51
- 中国癌症杂志 2019年29卷第7期页码：486-493
- 作者机构：
  
  1. 西安交通大学第二附属医院肿瘤放疗科,陕西,西安,710004
  2. 西安交通大学第二附属医院肿瘤科,陕西,西安,710004
- 作者简介：
- 基金信息：
  
  陕西省重点研发计划（2017SF-063）。
- DOI：10.19401/j.cnki.1007-3639.2019.07.002
  中图分类号： R735.1
- 网络出版：2019-07-12，
  
  纸质出版：2019-07-12
- 稿件说明：
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晋瑞,金迎迎,王敏聪,惠文涛,李毅,李芳,贾辉,潘继元,马红兵 . 抑制USP9x表达增强食管癌Ec9706-R细胞的放射敏感性[J]. 中国癌症杂志, 2019, 29(7): 486-493.

金迎迎, 王敏聪, 惠文涛, et al. Inhibition of USP9x expression enhances the radiosensitivity of esophageal cancer Ec9706-R cells51[J]. China Oncology, 2019, 29(7): 486-493.
晋瑞,金迎迎,王敏聪,惠文涛,李毅,李芳,贾辉,潘继元,马红兵 . 抑制USP9x表达增强食管癌Ec9706-R细胞的放射敏感性[J]. 中国癌症杂志, 2019, 29(7): 486-493. DOI： 10.19401/j.cnki.1007-3639.2019.07.002.

金迎迎, 王敏聪, 惠文涛, et al. Inhibition of USP9x expression enhances the radiosensitivity of esophageal cancer Ec9706-R cells51[J]. China Oncology, 2019, 29(7): 486-493. DOI： 10.19401/j.cnki.1007-3639.2019.07.002.

摘要

背景与目的：泛素特异性蛋白酶9x（ubiquitin-specific protease 9x，USP9x）与多种肿瘤的发生、发展以及肿瘤细胞的放射抗拒相关。研究发现，USP9x的表达与食管鳞状细胞癌的浸润深度和淋巴结转移相关，但其食管癌细胞放射抗拒作用尚未见报道。探究USP9x对放射抗拒食管癌Ec9706-R细胞放射敏感性的作用及其机制。方法：首先通过实时荧光定量聚合酶链反应（real-time fluorescent quantitative polymerase chain reaction，RTFQ-PCR）和蛋白质印迹法（Western blot）检测放射线照射后Ec9706-R细胞及其亲本细胞Ec-9706中USP9x和抗髓样细胞白血病-1（myeloid cell leukemia-1，Mcl-1）mRNA表达和蛋白水平。然后将Ec9706-R细胞随机分成3组：放射（irradiation，IR）组、IR+对照siRNA组（IR+si-NC组，转染Control siRNA）和IR+USP9x siRNA组（IR+si-USP9x组，转染USP9x siRNA），各组细胞均使用一定量的6 MV-X射线照射。噻唑蓝（methyl thiazolyl tetrazolium，MTT）比色法检测不同剂量（0、2、4、6和8 Gy）6 MV-X射线照射下各组细胞的活力。Transwell、流式细胞术、RTFQ-PCR和Western blot分别检测3组细胞在6 Gy照射下的细胞迁移、凋亡、Mcl-1 mRNA表达和蛋白水平以及增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）和DNA损伤修复相关基因核苷酸切除修复交叉互补基因1（excision repair cross-complementing gene 1，ERCC1）蛋白水平。结果：放射后Ec9706-R和Ec-9706细胞中USP9x和Mcl-1 mRNA表达和蛋白水平均增加，Ec9706-R细胞尤为显著（P0.05）。与IR组相比，IR+si-USP9x组中细胞活力、迁移细胞数目、PCNA、ERCC1和Mcl-1的表达均降低，细胞凋亡增加（P0.05）。但是与IR组相比，IR+si-NC组中上述指标均无显著变化（P0.05）。结论：抑制USP9x表达能够增强放射抗拒食管癌Ec9706-R细胞的放射敏感性，这可能是通过下调Mcl-1的表达发挥作用的。

Abstract

Background and purpose: Ubiquitin-specific protease 9x (USP9x) is related to the development and radioresistance of multiple cancers. Currently

it is reported that the USP9x expression is associated with invasive depth and lymphatic metastasis of esophageal squamous cancer. However

the effect of U5P9x on the radioresistance of esophageal cancer remains unknown. This study aimed to investigate the role of USP9x in the radiosensitivity of radioresistant esophageal cancer Ec9706-R cells and its mechanism. Methods: The mRNA expression and protein level of USP9x and myeloid cell leukemia-1 (Mcl-1) were detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot in Ec9706-R cells and the parental Ec9706 cells after irradiation. Then

Ec9706-R cells were randomly divided into 3 groups: irradiation (IR) group

IR+control siRNA group (IR+si-NC

transfected with control siRNA)

and IR+USP9x siRNA group (IR+si-USP9x

transfected with USP9x siRNA). All cells were exposed to designated dose of 6 MV-X irradiation. MTT analysis was used to determine the viability of cells following 6 MV-X irradiation with different doses (0

6 and 8 Gy). The migration

apoptosis and expression levels of Mcl-1

proliferating cell nuclear antigen (PCNA) and excision repair cross-complementing gene 1 (ERCC1) of Ec9706-R cells under 6 Gy irradiation were analyzed using Transwell

flow cytometry

real-time PCR and Western blot

respectively. Results: The mRNA expression and protein level of USP9x and Mcl-1 were elevated in both Ec9706-R and Ec9706 cells after irradiation

and their expression levels were more obvious in Ec9706-R cells (P0.05). Compared with the IR group

cell viability

migrated cell numbers and the expression levels of PCNA

Mcl-1 and ERCC1 were decreased

whereas cell apoptosis was increased in the IR+si-USP9x group (P0.05). However

compared with the IR group

these indexes were not changed in the IR+si-NC group (P0.05). Conclusion: Inhibition of USP9x enhances the radiosensitivity of esophageal cancer Ec9706-R cells

which may be dependent on the downregulation of Mcl-1.

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