放射耐受食管鳞癌细胞系基因表达谱研究

目的 构建放射耐受食管鳞癌细胞系,筛选放射耐受相关基因及探索耐受机制。方法 利用放射线反复照射食管癌细胞系KYSE410,构建放射耐受细胞系KYSE410-res。检测照射前后食管癌细胞增殖与凋亡情况,通过基因芯片技术检测放射线处理前后食管癌细胞基因表达差异情况,并对差异明显的基因进行验证。采用成组t检验。结果 KYSE410-res细胞较KYSE410细胞抗凋亡能力和增殖能力显著提高(P值均<0.05)。基因芯片结果显示KYSE410-res细胞中表达上调≥4倍基因有463个,下调≥4倍基因有251个。差异表达基因的功能主要集中在细胞增殖、粘附、信号转导、血管生成、活性氧代谢、细胞损伤修复及MAPK/ERK信号通路,OAS2和UBD是重要节点蛋白。荧光定量PCR法检测HLA-DQB1、MMP1、NCAM₁、ZNF521、GPC6、SELENBP1、LCN15、TFPI-2基因在KYSE410-res细胞中表达情况与基因芯片结果一致。结论 MAPK/ERK信号通路的异常激活、OAS2和UBD基因的表达上调、TFPI-2基因的表达下调伴随MMPs的表达上调,可能是食管癌细胞放射耐受的机制之一。

Gene expression profiling of a radioresistant esophageal squamous cancer cell line

Objective To establish a radioresistant esophageal squamous cancer cell line,and to identify the radioresistant genes and mechanisms.Methods The radioresistant KYSE410-res cell line was established by repeated exposure of cell line KYSE410 to radiation.The proliferation and apoptosis of esophageal squamous cancer cells were evaluated before and after radiation.The changes in gene expression of the esophageal squamous cancer cells after radiation were determined by gene microarray and analyzed by group t test.The genes with significant difference in expression after radiation were validated.Results The KYSE410-res cells had significantly enhanced proliferation and anti-apoptosis than the KYSE410 cells (all P＜0.05).The result of gene microarray showed that compared with the KYSE410 cells,the KYSE410-res cells had the expression of 463 and 251 genes upregulated and downregulated by no less than 4 folds,respectively.Those genes with different expression levels after radiation were mainly responsible for cell proliferation,adhesion,signal transduction,angiogenesis,reactive oxygen metabolism,cell damage repair,and the MAPK/ERK signaling pathway.OAS2 and UBD were key proteins in the network.In the KYSE410-res cells,the expression of HLA-DQBI,MMP1,NCAM1,ZNF521,GPC6,SELENBP1,LCN15,and TFPI-2 genes measured by real-time PCR was consistent with that measured by gene microarray.Conclusions Abnormal activation of the MAPK/ERK signaling pathway,upregulated expression of OAS2 and UBD,downregulated expression of TFPI-2,and upregulated expression of MMPs may play a role in radioresistance of esophageal cancer cells.