基因芯片表达谱筛选Kaposi肉瘤相关基因

中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (6): 403-406.

基因芯片表达谱筛选Kaposi肉瘤相关基因

王慧¹,吕国栋²,王晓东³,惠艳⁴,刘辉²,林仁勇⁵,王星⁵

1. 新疆医科大学第一附属医院
2.
3. 新疆医科大学第一附属医院皮肤科
4. 乌鲁木齐市新疆医科大学第一附属医院
5. 新疆医科大学第一附属医院医学研究中心

收稿日期:2010-08-18 修回日期:2010-12-06 出版日期:2011-06-15 发布日期:2011-06-02
通讯作者: 王星 E-mail:wangxing7610@yahoo.com.cn
基金资助:
新疆医科大学第一附属医院青年科研专项基金

Screening for Kaposi sarcoma-associated genes by using Genechip technology

Received:2010-08-18 Revised:2010-12-06 Online:2011-06-15 Published:2011-06-02

摘要/Abstract

摘要：

目的筛选Kaposi肉瘤相关基因。方法新疆本地1例经典型Kaposi肉瘤患者，提取病灶组织及正常皮肤组织的总RNA，逆转录成 cDNA，采用实时荧光定量PCR（RT-qPCR）的方法检测Kaposi肉瘤相关疱疹病毒（Kaposi sarcoma-associated herpersivirus，KSHV）基因K8.1、K2、ORF50在组织中的表达来确定所取标本有无交叉污染。随后进行荧光标记制备探针，与含有25100条人类基因的35K cDNA基因表达谱芯片进行杂交，用扫描仪扫描芯片荧光信号图像，并用软件对扫描图像进行数字化处理和分析。结果 RT-qPCR检测到 KSHV基因在Kaposi肉瘤病灶组织中的存在，而正常皮肤组织中未检测到，说明样本取材无交叉污染。在25 100个基因中，共有1313条差异基因，其中756条基因表达上调，557条基因表达下调。差异表达显著的基因涉及细胞凋亡、血管发生、细胞信号传导、蛋白加工和细胞周期调节等，如髓样细胞白血病-1基因（MCI-1）、膜联蛋白基因、丝氨酸蛋白酶抑制剂（SPINK5）基因等。结论 MCI-1、SPINK5等基因可能与Kaposi肉瘤的发病有关联。

关键词: 差异表达基因

Abstract:

Objective To screen for Kaposi sarcoma （KS）-related genes. Methods Tissue samples were obtained from the lesion and normal skin of a patient with KS in Xinjiang Uygur Autonomous Region, and total RNA was extracted from these samples and reverse transcribed into cDNA. Real-time fluorescent quantitative reverse transcription PCR （RT-qPCR） was performed to determine the expression of K8.1, K2 and ORF50 in these samples. The cDNA was labeled with fluorescein and hybridized to a human 35K genome array containing 25 100 genes. Subsequently, the signal images were scanned by a laser scanner and acquired images were analyzed by software. Results RT-qPCR revealed the mRNA expression of K8.1, K2 and ORF50 in the KS tissues but not in the normal skin tissues, indicating that there was no crossed contamination in these specimens. Among the 25 100 genes, 1313 genes were identified to be differentially expressed between KS and normal skin tissues, including 756 up-regulated genes and 557 down-regulated genes. These differentially expressed genes, such as myeloid cell leukemia-1 gene （MCI-1）, annexins （ANX） and serine proteinase inhibitor Kazal type 5 （SPINK5）, were associated with apoptosis, angiogenesis, cell signaling, protein processing, cell cycle regulation, and so on. Conclusion The differentially expressed genes such as MCI-1 and SPINK5 may be associated with the development of KS.

Key words: Differential expression gene

王慧吕国栋王晓东惠艳刘辉林仁勇王星. 基因芯片表达谱筛选Kaposi肉瘤相关基因[J]. 中华皮肤科杂志, 2011, 44(6): 403-406.

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