【作者】 郑华川；

【导师】 辛彦；

【作者基本信息】 中国医科大学 ， 肿瘤学， 2004， 博士

【摘要】 目的 观察胃癌中Kiss-1、基质金属蛋白酶-9(MMP-9)、FasL、Caspase-3和基质金属蛋白酶-7(MMP-7)表达及微血管密度(MVD)，探讨其在胃癌侵袭转移中的作用及相关分子病理学机制。 方法 采用PCR技术检测胃癌和相应正常胃黏膜中Kiss-1基因mRNA表达，利用Western印迹杂交检测相应组织中MMP-9蛋白表达，同时利用底物分解法检测了MMP-9酶活性，利用免疫组化检测FasL、Caspase-3和MMP-7蛋白在胃癌及相应正常胃黏膜中的表达，采用CD34免疫组化标记胃癌组织中微血管密度(MVD)，比较这些生物学指标与胃癌临床病理特征的关系，分析它们表达的相互关系。 结果 1．胃癌中Kiss-1 mRNA表达低于相应正常胃黏膜(P＜0.05)，而MMP-9蛋白表达相反(P＜0.05)。Kiss-1 mRNA表达与胃癌淋巴结转移呈负相关(P＜0.05)，肠型胃癌中Kiss-1基因mRNA表达水平高于弥漫型胃癌(P＜0.05)；MMP-9蛋白表达与胃癌淋巴结转移呈正相关(P＜0.05)。Kiss-1mRNA表达与MMP-9蛋白表达和酶活性均呈负相关(P＜0.05)；MMP-9蛋白表达与酶活性呈正相关(P＜0.05)。 2．胃原发灶癌细胞中FasL阳性表达率高于癌旁黏膜上皮细胞(P＜0.05)，Caspase-3则相反(P＜0.05)。70.8％原发灶中浸润淋巴细胞表达Caspase-3，阳性表达率显著高于原发灶癌细胞和癌旁上皮细胞(P＜0.05)。转移灶癌细胞中FasL阳性表达率低于相应原发灶癌细胞(P＜0.05)，Caspase-3则相反(P＜0.05)。原发灶癌细胞中FasL蛋白表达与胃癌肿块大小、浸润深度、转移、分化显著相关(P＜0.05)，与其浸润淋巴细胞中CasPase一表达呈正相关(P<0.05)，肠型胃癌中FasL蛋白表达高于弥漫型胃癌(P<0.05)。 3.胃癌癌旁粘膜上皮中MMP一阳性表达率为29 .2%，低于胃癌组织的阳性表达率69.0%(P<0.05);MMP一表达与胃癌肿块大小、浸润深度、转移和TNM分期呈正相关(P<0 .05);胃癌组织中MVD与胃癌肿块大小、浸润深度、转移和，rNM分期关系密切(P<0.05);胃癌组织中MVD的大小于MMP一表达成正相关(P<0.05)。结论 1.儿ss一1 mRNA表达下调以及MMP一蛋白表达上调在胃癌发生中起重要作用，异常Kiss一1 mRNA和MMP一表达与胃癌淋巴结转移密切相关，Kiss一1表达降低可能通过促进MMP一蛋白表达和提高其酶活性参与胃癌侵袭和转移。 2.FasL表达上调和CasPase一表达下调在胃癌发生中起重要作用。FasL可能通过诱导浸润淋巴细胞凋亡而参与胃癌分化、侵袭和转移，可作为一种反映胃癌病理生物学行为的有效指标。来自胃癌原发灶和转移灶的某种蛋白可能调节FasL和Caspase一表达来促进转移癌细胞凋亡水平和减弱转移癌细胞的免疫抵抗能力。 3.胃癌中MMP一表达上调，表达异常的MMP一可作为揭示胃癌病理生物学行为的客观指标，其可能通过参与胃癌生长、侵袭、转移和血管生成参与胃癌侵袭和转移过程。更多还原

【Abstract】 To observe Kiss-1, matrix metalloproteinase - 9 ( MMP-9 ) , FasL, Caspase-3, matrix metalloproteinase-7 ( MMP-7) expression and microvessel density ( MVD) in gastric cancer and explore their roles in progression of gastric cancer and their relevant molecular pathological mechanisms.MethodsKiss-1 mRNA expression was evaluated in gastric cancer and corresponding normal mucosa by RT-PCR. MMP-9 protein expression was examined by Western blot with its enzyme activity concerned. FasL, Caspase-3, MMP-7 expression and MVD were examined in gastric cancer or its corresponding normal mucosa by immunohistochemistry. These biological markers were compared with clinicopathological features of gastric cancer. The relationships between their expressions were analyzed as well.Results1. Kiss-1 mRNA expression level was lower in gastric cancer than that in normal gastric mucosa (P <0.05), whereas it was the converse for MMP-9 protein expression (P<0.05). Kiss-1 mRNA expression was negatively correlated with lymph node metastasis (P <0.05) ,whereas it was the converse for MMP-9 expression (P <0.05). Kiss-1 mRNA was less expressed in diffuse-type gastric cancers than that in intestinal-type ones (P <0.05). There was negative correlation trend between Kiss-1 mRNA expression and MMP-9 protein expression or enzymatic activity (P <0.05) , while positive correlation between MMP-9 pro-tein expression and its enzymatic activity (P <0.05).2. Primary cancer cells more frequently expressed FasL in gastric cancers than their adjacent epithelial cells ( P < 0. 05 ) , while it was the converse for Caspase-3 ( P < 0. 05 ). Infiltrating lymphocytes of primary foci showed strong immunoreactivity to Caspase-3 in 70. 8% of gastric cancers, more than their adjacent epithelial or cancer cells of primary foci (P <0. 05). FasL was less expressed in cancer cells of metastases than in those of corresponding primary foci ( P < 0. 05). It was the converse for Caspase-3 ( P < 0. 05). FasL expression was significantly correlated with tumor size, invasive depth, metastasis and differentiation in primary cancer cells (P <0. 05). FasL expression was higher in intestinal-type gastric cancers than that in diffuse-type ones (P <0. 05). FasL expression in primary gastric cancer cells paralleled positively with Caspase-3 expression in infiltrating lymphocytes of primary foci (P <0.05).3. MMP-7 showed positive expression in 29.2% of adjacent mucosa of gastric cancer, less than that in its gastric primary cancer (69.0% ). MMP-7 expression in primary foci was closely correlated with tumor size, invasive depth, metastasis and TNM staging of gastric cancer ( P < 0. 05). Positive correlation of mean MVD with tumor size, invasive depth, metastasis and TNM staging was found (P<0.05). Especially, mean MVD was dependent on MMP-7 expression in gastric cancer (P < 0. 05).Conclusions1. Down-regulated expression of Kiss-1 mRNA and up-regulated expression of MMP-9 protein were involved in gastric carcinogenesis. Altered expression of Kiss-1 mRNA and MMP-9 protein was closely correlated with lymph node metastasis of gatric cancer. Decreased expression of Kiss-1 mRNA was involved in invasion and metastasis of gastric cancer by enhancing protein expression and enzymatic activity of MMP-9.2. Up-regulated expression of FasL and down-regulated expression of Caspase-3 in primary cancer cells played an important role in gastric carcinogenesis. As an effective marker to reveal the pahobiological behaviors, FasL wasimplicated in differentiation, growth, invasion and metastasis of gastric cancer probably by inducing apoptosis of infiltrating lymphocytes. Protein derived from gastric primary cancer and metastatic microenvirongment might promote apoptosis of metastatic cells and eliminate immune counterattack by regulating Caspase-3 and FasL expression.3. Up-regulated MMP-7 expression played an important role in gastric car-cinogenesis. Altered expression of MMP-7 was an effective marker to reveal the pathobiological behaviors of gastric更多还原