摘要
目的 克隆人胃癌相关基因。方法 采用 5例胃窦部腺癌患者 (男 3例 ,女 2例 ,平均年龄48.8± 18.1岁 )配对的癌组织、癌旁组织及正常胃粘膜组织 ,提取其总 RNA,用荧光标记的差异显示技术(fluorescent differential display,FDD)分析同一患者的癌、癌旁与正常组织间表达基因的差异 ,将差异片段进行 Northern印迹、逆转录聚合酶链反应 (reverse transcription polymerase chain reaction,RT- PCR)、克隆、核苷酸序列分析 ,并将序列分析结果与 Gen Bank及 EST数据库进行同源性比较。结果 获得了一条712 bp的差异片段 ,在癌组织中的表达较正常与癌旁组织明显减弱 ,5例患者均有此特征 ,命名为 W4。Northern印迹显示与 FDD的结果一致。BL AST分析表明 ,该序列与 Gen Bank和 EST库所存数据无同源性 ,Gen Bank登录号为 AF15 0 6 31。RT- PCR显示 11/ 15例胃腺癌组织表达降低或无表达。结论 发现了一条与胃腺癌相关的新的 c
Objective To identify human gastric cancer related genes. Methods Specimens of paired tumor, paratumor and normal gastric mucosa tissues were collected from five patients (male 3, female 2, with average age 48 8±18 1 years) who suffered from stomach antrum adenocarcinoma. Total RNA samples were extracted from these specimens, then studied by fluorescent differential display reverse transcription polymerase chain reaction (DDRT PCR) analysis. The differentially expressed bands of interest were recovered, purified and cloned, then they were analyzed by sequencing, Northern blot and RT PCR. Through BLAST, the sequencing results were compared with GenBank database for homology analysis.Results One of the interesting cDNA bands expressed much lower in all five tested tumor samples than in their normal and paratumor counterparts. This band was named W4. Northern blot analysis showed a consistent result with that of DDRT PCR. W4 was cloned into pGEM T easy vector. Sequence analysis showed that W4 consists of 712bp, this sequence was named W44.BLAST analysis revealed that W44 has extremely low sequence identity with any genes from GenBank and any sequences from EST database. This sequence data was submitted to GenBank with accession No. AF150631. RT PCR analysis showed that W4 was expressed much lower in 11/15 gastric cancer tissue than in paratumor and normal samples. Conclusion A novel cDNA sequence related to human stomach adenocarcinoma was identified.
出处
《中华医学遗传学杂志》
CAS
CSCD
2001年第1期43-46,共4页
Chinese Journal of Medical Genetics
基金
军队"九五"医药卫生科研基金(96M145)
关键词
胃癌
腺癌
基因
差异显示
逆转录聚合酶链反应
stomach cancer
adenocarcinoma
gene
differential display
reverse transcription poly merase chain reaction
