基质金属蛋白酶-9及其mRNA在胃癌中的表达与血管新生的关系

目的　观察基质金属蛋白酶 9(MMP 9)与MMP 9mRNA在胃癌组织及细胞中的表达与作用特点 ,及其与肿瘤浸润、转移及血管新生之间的关系。方法　利用 74例胃癌标本及体外培养人胃低分化腺癌细胞BGC82 3,经连续切片免疫组织化学染色、原位杂交、侵袭实验、酶谱分析及形态定量分析方法 ,进行金属蛋白酶 (MMP) 9、MMP 9mRNA、CD34、α SMA、VEGFR 1、VEGFR 2的检测 ,并结合临床病理参数进行综合分析。结果　MMP 9及其mRNA的表达均与癌组织的浸润深度、分化程度、淋巴结转移密切相关 ;MVD与癌组织的浸润深度、淋巴结转移关系密切 ;BGC82 3细胞MMP 9,MMP 9mRNA ,VEGFR 2染色均为阳性 ,VEGFR 1染色阴性 ;MMP 9抗体可使BGC82 3细胞侵袭细胞数由 6 9± 5个 /40 0倍下降为 (18± 4 )个 /40 0倍 ;直接培养于培养皿表面的BGC82 3细胞有稳定的MMP 9,MMP 2酶原表达 ,但培养于I型胶原包被培养皿表面的BGC82 3细胞MMP 2表达明显增强并出现活化形式 ;BGC82 3细胞与VEGF共孵育 ,条件培养基中MMP 9酶原含量以VEGF剂量依赖的方式增加。结论　胃癌细胞自身具有产生与分泌MMP 9的能力 ,MMP 9对肿瘤的浸润、转移及间质血管新生具有促进作用。

Expression of MMP-9 and MMP-9 mRNA in gastric carcinoma and its correlation with angiogenesis

OBJECTIVE: To investigate the expression of MMP-9 and MMP-9 mRNA in the tissue and cell line of the gastric carcinoma and its correlation with invasion, metastasis and angiogenesis of the cancer. METHODS: Seventy-four specimens of resected human gastric carcinoma were collected. Human lowly differentiated gastric carcinoma cells of BGC823 line were cultured. Hematoxylin and eosin staining, immunohistochemistry, in situ hybridization, and zymography were used to examine the expression of MMP-9, MMP-9 mRNA, CD34, alpha-SMA, VEGFR-1, VEGFR-2. Invasion test was made to observe the number of cells penetrating the Matrigel. The relationship between the expression of MMP-9 and MMP-9 mRNA and the clinical parameters was analyzed. RESULTS: The positive rate of MMP-9 expression was 77.03% in gastric carcinoma tissues (57/74). Most of the MMP-9 positive cells were located at the invasive areas. The average microvascular density (MVD) of the 74 cases of gastric carcinoma was 73.76 +/- 11.50/200 x visual field. MVD was 75.46 +/- 10.98 in the MMP-9 positive group and was 68.08 +/- 11.71 in the MMP-9 negative group (P < 0.05). The positive rate of MMP-9 mRNA expression was 72.97% in the gastric carcinoma tissues (54/74). The expressions of MMP-9 and its mRNA were closely related to invasion, differentiation and lymph nodes status of the gastric carcinoma; MVD was closely related to the invasion and lymph nodes status of the gastric carcinoma. All BGC823 cells were MMP-9, MMP-9 mRNA, VEGFR-2 positive and VEGFR-1 negative. The number of invasive BGC823 cells was (18 +/- 4)/400 x in the MMP-9 antibody group and was (69 +/- 5)/400 x in the control group. BGC823 cells cultivated in culture dishes expressed the proenzyme of MMP-2 and MMP-9 stably. BGC823 cells cultivated on the surface of type I collagen expressed more MMP-2 proenzyme and active MMP-2. When BGC823 cells were incubated with VEGF, the content of MMP-9 proenzyme was increased by VEGF dose-dependently. CONCLUSION: Gastric cancer cells have the ability to produce and secret MMP-9 which can promote the invasion and metastasis of the neoplasm.