涎腺腺样囊性癌高、低转移细胞系基因表达谱及基质金属蛋白酶表达差异

Analysis of differential expression genes related to different metastasis potential of adenoid cystic carcinoma using restriction fragments differential display PCR

目的筛选与涎腺腺样囊性癌转移相关的候选基因,并对其中的候选基因进行初步的验证。方法用限制片段差异显示PCR技术(restriction fragments differential display PCR,RFDD-PCR)建立涎腺腺样囊性癌高、低转移细胞株(ACC-M、ACC-2)的表达谱。对两个表达谱的片段进行比较,通过生物信息学的分析,初步筛选出候选基因。用半定量逆转录PCR技术对筛选出的基因进行初步验证。结果 RFDD-PCR方法共获得5420个基因片段,其中包含12个基质金属蛋白酶(matrix metalloproteinase ,MMP)基因。半定量逆转录PCR方法发现MMP2、MMP7、MMP9、MMP14、MMP15、MMP24在ACC-M和ACC-2中的表达存在明显差异。结论构建了ACC-M和ACC-2细胞株的表达谱,为寻找目的基因奠定了基础。发现MMP2、MMP7、MMP9和MMP15与涎腺腺样囊性癌的发生、发展、转移有关,不同肿瘤细胞的转移能力可能与不同的MMPs家族基因相关。

Objective To construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma. Methods Restriction fragroents differential display PCR （RFDD-PCR） was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were 〉creened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR（RT-PCR）. Results Two gene expression profiles including 5420 gene fragments were , constrocted, 12 genes of a family called matrix metalloproteinase genes （MMPs） were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR .also identified this different expression of MMP2, MMP7,MMP9,MMP14,MMP15 and MMP24. Conclusion The construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2, MMP7, MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.