SWI/SNF复合体基因突变促进NSCLC细胞在NSI小鼠体内肝转移的研究  

SWI/SNF Complex Gene Mutations Promote the Liver Metastasis of Non-small Cell Lung Cancer Cells in NSI Mice

作　　者：高玲玲 谢至 林首恒 吕志异 周文斌 陈冀 朱琳琳[1,2,3] 张莉 曾鹏辉[1,2,3] 黄晓丹 颜文青 陈宇[3] 卢丹霞 张水莲 郭伟浜[3] 李鹏 张绪超 Lingling GAO;Zhi XIE;Shouheng LIN;Zhiyi LV;Wenbin ZHOU;Ji CHEN;Linlin ZHU;Li ZHANG;Penghui ZENG;Xiaodan HUANG;Wenqing YAN;Yu CHEN;Danxia LU;Shuilian ZHANG;Weibang GUO;Peng LI;Xuchao ZHANG(Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510080,China;The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510515,China;Guangdong Provincial Key Laboratory of Translational Medicine in Lung Cancer,Medical Research Center,Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510080,China;Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,China;School of Medicine,South China University of Technology,Guangzhou 510006,China)

机构地区：[1]南方医科大学附属广东省人民医院(广东省医学科学院),广州510080 [2]南方医科大学第二临床医学院,广州510515 [3]南方医科大学附属广东省人民医院(广东省医学科学院),医学研究部,广东省肺癌转化医学重点实验室,广州510080 [4]中国科学院广州生物医药与健康研究院,广州510530 [5]华南理工大学医学院,广州510006

出　　处：《中国肺癌杂志》2023年第10期753-764,共12页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金项目(No.82173202)资助

摘　　要：背景与目的SWI/SNF复合体(switch/sucrose nonfermentable chromatin-remodeling complex,SWI/SNF)是一种重要的染色质重塑复合物,其亚基变异在多种肿瘤中存在,并与多种肿瘤细胞生物学特征相关。但其基因突变是否参与非小细胞肺癌(non-small cell lung cancer,NSCLC)肝转移过程尚不清楚。本研究拟探究SWI/SNF复合体基因突变对NSCLC肝转移的影响及潜在机制。方法我们使用全外显子组测序(whole-exome sequencing,WES)分析了NSCLC细胞H1299、H23和H460中SWI/SNF复合体基因突变。通过CRISPR/Cas9(clustered regularly interspaced short palindromic repeats)技术构建了ARID1A基因稳定敲除的H1299细胞株,建立了小鼠模型模拟NSCLC肝转移,观察不同基因突变对肝转移的影响。利用RNA-Seq和蛋白印迹分析差异基因的表达,并通过免疫组化技术(immunohistochemistry,IHC)检测了SWI/SNF复合体调控的靶分子在小鼠肝转移灶中的表达。结果WES分析确定了SWI/SNF复合体基因的突变情况。动物实验结果显示SWI/SNF复合体基因突变与免疫缺陷小鼠较高的肝转移率相关。转录组测序和蛋白印迹分析显示SWI/SNF复合体基因突变细胞中ALDH1A1和APOBEC3B表达上调,尤其是ARID1A蛋白缺失的H460和H1299 sgARID1A中ALDH1A1表达水平显著上升。IHC染色亦显示H460和H1299 sgARID1A细胞肝转移灶中ALDH1A1高表达。结论本研究强调了SWI/SNF复合体基因ARID1A和SMARCA4等突变在促进肺癌细胞肝转移中的关键作用。这些基因突变可能通过促进ALDH1A1与APOBEC3B高表达进而发挥肝特异性转移的作用,为深入探究肺癌肝转移分子机制提供了新线索。Background and objective The switch/sucrose nonfermentable chromatin-remodeling(SWI/SNF)complex is a pivotal chromatin remodeling complex,and the genomic alterations(GAs)of the SWI/SNF complex are observed in several cancer types,correlating with multiple biological features of tumor cells.However,their role in liver metastasis of non-small cell lung cancer(NSCLC)remains unclear.Our study aims to investigate the role and potential mechanisms underlying NSCLC liver metastasis induced by the GAs of SWI/SNF complex.Methods The GAs of SWI/SNF complex in NSCLC cell lines(H1299,H23 and H460)were identified by whole-exome sequencing(WES).ARID1A knockout H1299 cell was constructed with the CRISPR/Cas9 technology.The mouse model of liver metastasis from NSCLC was established to simulate lung cancer liver metastasis and observe the metastasis rate under different gene mutation conditions.RNA sequencing and Western blot were conducted for differential gene expression analysis.Immunohistochemistry(IHC)analysis was used to assess protein expression levels of SWI/SNF-regulated target molecules in mouse liver metastases.Results WES analysis revealed intracellular gene mutations.The animal experiments demonstrated a correlation between the GAs of SWI/SNF complex and a higher liver metastasis rate in immunodeficient mice.Transcriptome sequencing and Western blot analysis showed upregulated expression of ALDH1A1 and APOBEC3B in SWI/SNF-mut cells,particularly in ARID1A-deficient H460 and H1299 sgARID1A cells.IHC staining of mouse liver metastases further demonstrated elevated expression of ALDH1A1 in the H460 and H1299 sgARID1A group.Conclusion This study underscores the critical role of the GAs of SWI/SNF complex,such as ARID1A and SMARCA4,in promoting liver metastasis of lung cancer cells.The GAs of SWI/SNF complex may promote liver-specific metastasis by upregulating ALDH1A1 and APOBEC3B expression,providing novel insights into the molecular mechanisms underlying lung cancer liver metastasis.

关 键 词：肺肿瘤 SWI/SNF复合体 突变 肿瘤转移 

分 类 号：R734.2[医药卫生—肿瘤]