结直肠癌原发瘤细胞和淋巴结转移瘤细胞基因表型差异的研究

Heterogeneity between primary colorectal carcinoma and matched metastases as revealed analysis of cDNA microarray

目的:研究结直肠癌转移过程中基因表型的变化,深入了解肿瘤转移发生机制。方法:获取新鲜肿瘤原发灶和转移灶组织,种植于裸鼠皮下,2周后取出肿瘤,观察两个部位肿瘤的生物学特点。抽提人原发灶和转移灶组织RNA,与人类全基因组基因芯片杂交,筛查转移相关基因,并行荧光定量RT-PCR鉴定。结果:转移灶癌细胞生长迅速,穿透薄膜,浸润入周围组织,而原发灶癌细胞有完整包膜。基因芯片杂交发现转移灶较原发灶癌细胞高表达基因有63条,而低表达基因有160条。其中MTA1、KISS1、MTSS1、S100A4、DAF、CLDN2、CA12、GalNAc-T3、RNASET2、ANXA10、ANXA1、KRT20、Grb2、LCK、EPHB4、CyclinL1、CTSD、BMP7和CDK6等已证实与转移的发生明确相关。经RT-PCR证实结果可靠。结论:转移灶癌细胞浸润和转移能力要高于起源灶细胞。肿瘤的转移是多基因参与的结果,这对于肿瘤转移机制的认识和防治有重要意义。

Objective：To investigate the mechanisms involved in metastasis of colorectal carcinoma by detecting changed expression of genes during the progression of metastasis. Methods： Fresh tissues from primary tumors and lymph node metastases were implanted with nude mice. Two weeks later, characteristics of two sites were analyzed. Tumor tissues from two sites were reversed into total mRNA and then into cDNA. The cDNAs were hybridized with human genomic cDNA microarrays. RT-PCR was used to confirm the results of the cDNA microarray. Results：Metastatic cancer cells grew more quickly than primary cancer cells and penetrated into surrounding tissues through membrane basement. While primary cancer cells had complete membrane basement. Sixty-three genes were up-regulated and 160 genes were down-regulated in metastatic cells than in primary cancer cells. Some of these genes, including MTA1, KISS1, MTSS1,S100A4, DAF, CLDN2, CA12, GalNAc-T3, RNASET2, ANXA10, ANXA1, KRT20, Grb2, LCK,EPHB4, cyclin L1, CTSD, BMP7, and CDK6, have been previously associated with metastasis. Conclusion：Metastatic cancer cells have a higher ability of metastasis potential than primary cancer cells. Multiple genes contribute to the progression of metastasis of colorectal carcinoma, which may be the target of treating this disease.