摘要
目的探讨应用基因芯片技术筛选食管癌侵袭转移相关基因的相关机制。方法利用Transwell侵袭小室技术建立具有不同侵袭转移潜能的人食管鳞癌Eca109和Eca109-T4细胞株;分别提取Eca109和Eca109-T4的总RNA,用转录Cy5和Cy3荧光标记成c DNA探针,再与Affymetrix基因芯片杂交,杂交信号用GenechipScanner 3000扫描,SAM 3.0软件分析和处理数据;运用实时荧光定量PCR(qRT-PCR)技术对CXCR7、SDC2、HSP90α1、TNFRSF10D基因进行验证。结果筛选出Eca109和Eca109-T4差异基因有80个:与母系比较,在Eca109-T4中上调表达的基因有34个,下调表达的基因有46个。经qRT-PCR验证:CXCR7在Eca109-T4中的mRNA表达量(9.86±1.38)显著高于Eca109(6.04±1.88,P<0.05),SDC2在Eca109-T4中的mRNA表达量(0.07±0.006 7)显著高于Eca109(0.04±0.003 7,P<0.05),HSP90α1在Eca109-T4中的mRNA表达量(0.18±0.046 0)显著高于Eca109(0.05±0.009 8,P<0.05),TNFRSF10D在Eca109-T4中的mRNA表达量(0.009±0.000 6)与Eca109(0.005±0.004 0,P>0.05)无明显差异。结论基因芯片技术是一高效筛选食管癌侵袭转移相关基因的方法,CXCR7、SDC2、HSP90α1等基因可能与食管癌侵袭转移机制相关。
Objective To screen esophageal cancer invasion and metastasis associated genes to understand esophageal squamous carcinoma invasion and metastasis related mechanisms by gene chip technology. Methods Transwell invasion chamber technology was used to establish human esophageal squamous carcinoma Eca109 and Eca109-T4 cell lines with different invasion and metastasis ability. Eca109 and Eca109-T4 total RNA were extract,ed and reversed transcribed into c DNA probes which labeled with cy5 and Cy3 fluorescence,then it were hybridized by Affymetrix gene chip. Genechip-Scanner 3000 scan hybridization signal was used,and CXCR7,SDC2,HSP90α1 and TNFRSF10 D genes were analyzed by real-time polymerase chain reaction( qRT-PCR).Results 80 differentially expressed genes were identified between Eca 109 and Eca109-T4.Compared to Eca109,34 of 80 differentially expressed genes were upregulated and the remaining 46 were downregulated. qRT-PCR showed that the expressions of CXCR7,SDC2,HSP90α1 at mRNA level were significantly higher in Eca109-T4( 9.86±1.38,0.07±0.0067,0.18±0.046 0) than thatose in Eca109( 6.04±±1.88,0.04±0.003 7,0.05±0.009 8,P<0.05). But there was no obvious difference in the expression of TNFRSF10 D gene between Eca109-T4( 0.009±0.000) and Eca109( 0.005±0.004 0,P>0. 05).Conclusions Gene chip technology is a high efficiency method to screen esophageal cancer metastasis related gene. CXCR7,SDC2 and HSP90α1 may be associated with esophageal cancer metastasis and invasion mechanism.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2015年第8期2148-2152,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金(No.81001101)
新疆维吾尔自治区自然科学基金(No.2010211B20)
关键词
食管癌
转移
基因芯片
差异基因
Esophageal cancer
Metastasis
Gene chip
Genetic variations
