中华皮肤科杂志 ›› 2004, Vol. 37 ›› Issue (2): 65-67.

• 论著 •    下一篇

运用cDNA芯片技术对恶性黑素瘤相关基因的研究

林彤, 孙建方   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042
  • 收稿日期:2003-03-31 出版日期:2004-02-15 发布日期:2004-02-15

Study on Melanoma Gene Expression Profile by cDNA Microarray

LIN Tong, SUN Jian-fang   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences, Peking Union Medical College, Nanjing 210042, China
  • Received:2003-03-31 Online:2004-02-15 Published:2004-02-15

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摘要： 目的 运用cDNA芯片技术研究恶性黑素瘤(MM)相关基因。方法 提取并分离得到MM及正常人皮肤组织的mRNA,逆转录成cDNA,以33P标记,再与含有2000条人类基因的微阵列杂交,获得MM的基因表达谱,并对有兴趣的共同差异表达基因进行实时定量RT-PCR验证。结果 2倍,5倍及10倍以上差异表达基因分别占4.7%~6.15%,0.75%~1.4%和0.45%~0.5%。这些基因主要属于原癌和抑癌、凋亡及细胞周期等多种肿瘤相关基因。4例MM的共同差异表达基因有3条,均呈表达下降。对其中组氨酸三聚体核苷结合蛋白基因(HINT),视网膜母细胞瘤结合蛋白1样基因(BCAA)进行实时定量RT-PCR验证,结果也显示HINT及BCAA在MM中低表达。结论 基因芯片技术是研究MM发生、发展机制的有效手段,首次发现HINT和BCAA两条基因在MM的中表达下降,需要对其确切的致病机制及表达特异性作进一步的研究。

关键词: 黑色素瘤, 寡核苷酸序列分析, 逆转录聚合酶链反应

Abstract: Objective To study the expression of malignant melanoma(MM) related genes by cDNA microarray technique. Methods mRNA, extracted from tissues of patients and normal controls, was reversely transcripted into cDNA and marked with 33P. The cDNA probes were hybridized to cDNA microarrays, which contained 2000 human genes each array. The down-regulation of two co-differentiated expressed genes was confirmed by quantitative real-time RT-PCR. Results Different expression between MM and normal controls was found in 4.7%-6.15% of genes by more than 2 times,0.75%-1.4% by more than 5 times, and 0.45-0.5% by more than 10 times. These genes were pro-oncogenes, tumor suppressor genes, genes related to apoptosis, cell cycle related genes, and so on. Three genes were down-regulated in all of the patients. Two of those genes, histidine triad nucleotide-binding protein (HINT) and RBP1-like protein (BCAA), were down-regulated, as identification by quantitative real-time RT-PCR. Conclusions cDNA microarray can be used effectively to reveal melanoma gene expression profiling for the propose of carcinogenesis study. HINT and BCAA are the first reported genes down-regulated in MM. However, further studies are needed for their expressive specificity and mechanism in MM.

Key words: Melanoma, Oligonucleotide array sequence analysis, Reverse transcriptase polymerase chain reaction