应用低密度芯片初步筛选胃癌淋巴转移相关基因的研究

目的:运用低密度cDNA芯片技术初步筛选影响胃癌淋巴结转移的分子标志。方法:运用自制低密度cDNA芯片检测15例胃癌患者原发癌灶和淋巴结转移癌中多个基因mRNA表达,并用RT-PCR方法进行验证,分析其与胃癌侵袭和淋巴结转移的相关性。结果:hTERT在淋巴结转移癌中表达明显高于胃原发癌灶(P= 0．001)。原发癌灶中MMP-7和Heparanase表达在小结节孤立型淋巴结病例中明显高于大结节融合型者(P值分别为0．022和0．002),而CDH1表达明显降低(P= 0．002);淋巴结转移癌灶中CDH1表达在小结节孤立型淋巴结病例中亦明显降低(P=0．046)。按淋巴结转移个数分成轻重两组(≤6个为轻度转移,≥7个为重度转移),原发癌灶中MMP-7和hTERT表达在重组明显高于轻组(P值分别为0．001和0．005);淋巴结转移癌灶中MMP-7、S100A4和hTERT基因表达在重组明显增高(P值分别为0．000 05、0．007和0．016),而nm23H1和CDH1表达明显降低(P值分别为0．013和0．001)。胃原发癌中MMP-7、Heparanase、S100A4过表达和(或)nm23H1、CDH1、KAI-1表达降低或缺失者,胃癌多侵透浆膜,且浆膜受侵面积较大。结论:hTERT、MMP-7、S100A4、Heparanase、CDH1和nm23H1基因表达与胃癌淋巴结转移关系密切,可望成为胃癌淋巴结转移诊断和治疗的分子靶点。

Elementary screening lymph node metastatic-related genes in gastric cancer by low-density cDNA microarray

OBJECTIVE: To screen the lymph node metastatic-related genes in human gastric cancer hy the low-density cDNA microarray technique. METHODS: A total of 15 gastric cancer primary and nodal involvement lesions were examined by a low-density cDNA microarray containing 23 genes and RT-PCR was used for the further verification. The correlation between the gene expression and invasion and nodal involvement were analyzed. RERULTS:Hybridization signals bad good specificity, which were seen for the house keeping genes (internal control) . but were not seen for the hepatitis B gene (negative control) and spotting solution only (blank control). The expressions of hTERT in the nodal lesions were higher than those in the coupled primary lesions (P=0. 001). With respect to the number of nodal involvement, MMP-7 and hTERT expressions in the primary lesions were higher in the heavier nodal involvement group (no less than 7) than those in the lighter one (no more than 6)(P=0. 001,0. 005 respectively). MMP-7, S100A4 and hTERT expressions in nodal lesions were higher in the heavier than those in the lighter (P=0. 000 05,P = 0. 007,P=0. 016, respectively), whereas nm23H1 and CDH1 were lower(P = 0. 013,P = 0. 001). With respect to the nodal type, in the primary lesions MMP-7 and Heparanase expressions in the small node isolation were higher than those in the big node fusion (P = 0. 022,0. 002 respectively). Whereas CDH1 expression was lower (P=0. 002), so did CDH1 expression in nodal lesions (P = 0. 046). In patients with MMP-7, Heparanase, S100A4 overexpressions and/or nm23H1, CDH1, KAI-1 lessexpressions or loss in gastric cnacer primary lesions, most cases were serosa involvement and the area of serosa involvement was biggish. CONCLUSION: MMP-7, hTERT, Heparanase, CDH1 correlated closely with nodal involvement in gastric carcinomas, and may be the molecular target for an early diagnosis of nodal involvement and treatment in gastric cancer.