中国老年学杂志
中國老年學雜誌
중국노년학잡지
CHINESE JOURNAL OF GERONTOLOGY
2015年
8期
2148-2152
,共5页
张琪琪%张志强%温浩%李秀娟
張琪琪%張誌彊%溫浩%李秀娟
장기기%장지강%온호%리수연
食管癌%转移%基因芯片%差异基因
食管癌%轉移%基因芯片%差異基因
식관암%전이%기인심편%차이기인
Esophageal cancer%Metastasis%Gene chip%Genetic variations
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cDNA探针，再与Affymetrix基因芯片杂交，杂交信号用 Genechip? Scanner 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±0.0067）显著高于 Eca109（0.04±0.0037，P＜0.05），HSP90α1在 Eca109-T4中的 mRNA 表达量（0.18±0.0460）显著高于Eca109（0.05±0.0098，P＜0.05），TNFRSF10D在Eca109-T4中的mRNA表达量（0.009±0.0006）与 Eca109（0.005±0.0040，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。
目的：探討應用基因芯片技術篩選食管癌侵襲轉移相關基因的相關機製。方法利用Transwell侵襲小室技術建立具有不同侵襲轉移潛能的人食管鱗癌Eca109和 Eca109-T4細胞株；分彆提取Eca109和Eca109-T4的總RNA，用轉錄Cy5和Cy3熒光標記成cDNA探針，再與Affymetrix基因芯片雜交，雜交信號用 Genechip? Scanner 3000掃描，SAM 3.0軟件分析和處理數據；運用實時熒光定量 PCR（qRT-PCR）技術對 CXCR7、SDC2、HSP90α1、TNFRSF10D基因進行驗證。結果篩選齣Eca109和Eca109-T4差異基因有80箇：與母繫比較，在Eca109-T4中上調錶達的基因有34箇，下調錶達的基因有46箇。經qRT-PCR驗證：CXCR7在Eca109-T4中的mRNA錶達量（9.86±1.38）顯著高于Eca109（6.04±1.88，P＜0.05），SDC2在Eca109-T4中的mRNA錶達量（0.07±0.0067）顯著高于 Eca109（0.04±0.0037，P＜0.05），HSP90α1在 Eca109-T4中的 mRNA 錶達量（0.18±0.0460）顯著高于Eca109（0.05±0.0098，P＜0.05），TNFRSF10D在Eca109-T4中的mRNA錶達量（0.009±0.0006）與 Eca109（0.005±0.0040，P＞0.05）無明顯差異。結論基因芯片技術是一高效篩選食管癌侵襲轉移相關基因的方法，CXCR7、SDC2、HSP90α1等基因可能與食管癌侵襲轉移機製相關。
목적：탐토응용기인심편기술사선식관암침습전이상관기인적상관궤제。방법이용Transwell침습소실기술건립구유불동침습전이잠능적인식관린암Eca109화 Eca109-T4세포주；분별제취Eca109화Eca109-T4적총RNA，용전록Cy5화Cy3형광표기성cDNA탐침，재여Affymetrix기인심편잡교，잡교신호용 Genechip? Scanner 3000소묘，SAM 3.0연건분석화처리수거；운용실시형광정량 PCR（qRT-PCR）기술대 CXCR7、SDC2、HSP90α1、TNFRSF10D기인진행험증。결과사선출Eca109화Eca109-T4차이기인유80개：여모계비교，재Eca109-T4중상조표체적기인유34개，하조표체적기인유46개。경qRT-PCR험증：CXCR7재Eca109-T4중적mRNA표체량（9.86±1.38）현저고우Eca109（6.04±1.88，P＜0.05），SDC2재Eca109-T4중적mRNA표체량（0.07±0.0067）현저고우 Eca109（0.04±0.0037，P＜0.05），HSP90α1재 Eca109-T4중적 mRNA 표체량（0.18±0.0460）현저고우Eca109（0.05±0.0098，P＜0.05），TNFRSF10D재Eca109-T4중적mRNA표체량（0.009±0.0006）여 Eca109（0.005±0.0040，P＞0.05）무명현차이。결론기인심편기술시일고효사선식관암침습전이상관기인적방법，CXCR7、SDC2、HSP90α1등기인가능여식관암침습전이궤제상관。
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 Eca 109 and Eca109-T4 cell lines with different invasion and metastasis ability .Eca109 and Eca109-T4 total RNA were extract,ed and reversed transcribed into cDNA probes which labeled with cy 5 and Cy3 fluorescence,then it were hybridized by Affymetrix gene chip .Genechip?-Scanner 3000 scan hybridization signal was used , and CXCR7,SDC2, HSP90α1 and TNFRSF10Dgeneswereanalyzedbyreal-timepolymerasechainreaction(qRT-PCR).Results 80differentiallyexpressedgeneswereiden-tified 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 CXCR 7,SDC2,HSP90α1 at mRNA level were significantly higher in Eca 109-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 TNFRSF 10D 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 .
