新型丝氨酸蛋白酶SNC19(ST14)基因编码蛋白在大肠癌中的表达

目的 研究新型丝氨酸蛋白酶在 SNC19大肠癌中的表达分布与临床病理资料的相关性及其促分化功能的重要意义。方法 应用 RT- PCR方法进行 SNC19在 39对正常大肠黏膜和肿瘤组织的 RNA水平表达的检测。应用免疫组化法研究 5 3例大肠肿瘤组织中 SNC19蛋白的表达分布情况。结果 39例正常大肠黏膜及相应肿瘤组织中均有 SNC19m RNA表达。正常黏膜组的 SNC19m RNA表达水平要高于肿瘤组 ,差异有显著性。表达水平高低与各个临床指标无明显关系。SNC19蛋白主要分布在大肠黏膜表面上皮 ,而腺体上皮表达为阴性。高分化肠癌中SNC19蛋白表达阳性率为 74 .2 % ( 2 3/ 31) ,中分化肠癌中 SNC19蛋白表达阳性率为 5 7.1% ( 8/ 14 ) ,低分化肠癌中SNC19蛋白表达阳性率为 12 .5 % ( 1/ 8) ,差异有显著性 ( P<0 .0 1)。SNC19蛋白表达在肿瘤分期和淋巴结转移的差异无显著性。结论 SNC19蛋白在大肠癌中表达与促分化有关 。

新转移相关蛋白SNC19翻译后加工及其活性形式的研究

目的 :进一步从蛋白水平研究 SNC19编码蛋白与肿瘤转移的关系 ;方法 :免疫印迹法检测该蛋白在不同肿瘤细胞系及大肠癌组织中的表达 ;构建 Psectag2 A- SNC19(ORF)真核表达质粒 ,转染至乳腺癌细胞 BCAP37中瞬时表达 ,产物经亲和层析纯化 ,用 anti- His m Ab进行 Western blot鉴定其加工形式 ,并观察不同的加工产物是否具有明胶酶活性。结果 :免疫印迹法显示 ,在所选肿瘤细胞系和组织中 SNC19蛋白存在差异表达及翻译后加工 ;转染的 BCAP37细胞产生 12 0 k D和 75 k D两种形式的 SNC19蛋白 ,其中只有 75 k D的形式被证明具有明胶酶活性。结论 :SNC19蛋白在不同的肿瘤细胞系和肿瘤组织中存在差异表达及翻译后加工 ,其中 75 k D的形式被证明具有明胶酶活性。

新型丝氨酸蛋白酶SNC19在不同细胞系中表达及其对细胞生物学行为的影响

目的 :研究新型丝氨酸蛋白酶SNC19编码蛋白的功能及可能在肿瘤转移过程中的作用。方法 :制备针对它胞外区的单克隆抗体 ;免疫印迹法检测在不同细胞系中该蛋白的表达 ;Northernblot检测在不同上皮来源细胞系中其mRNA的表达 ;用单抗阻断法研究其对细胞迁移能力和细胞粘附能力的影响。结果 :自制抗体做West ernblot显示在BCAP37、COLO2 0 5、SW4 80中该蛋白存在两种形式 ,大小为 12 0kD和 6 0kD左右 ,而在SW6 2 0中只存在 6 0kD一种形式 ;Northernblot显示该蛋白在大部分细胞系中均有较高表达 ,大小为 3 4kd左右 ,mRNA水平只有一种形式 ,而在卵巢癌细胞系OV、舌癌细胞系TCA8113、口腔表皮样癌细胞系KB及胃癌细胞系SGC790 1中未检测到其表达 ;单抗阻断法发现实验组SW4 80 (anti-SNC19mAb 2 0mg/L)迁移速度明显慢于对照组 ,在SW4 80 /SW4 80同质粘附和SW4 80 /NIH3T3异质细胞粘附实验中 ,在实验的初始阶段 ,实验组SW4 80 (anti -SNC19mAb 2 0mg/L) /SW4 80、SW4 80 (anti-SNC19mAb 2 0mg/L) /NIH3T3粘附率明显大于对照组 ,但随着时间的推移这种差异逐渐缩小。结论 :SNC19编码蛋白与同质、异质细胞之间的粘附以及细胞的迁移运动密切相关。作为一种新型丝氨酸蛋白酶 ,SNC19编码蛋白可能与细胞迁移、组织重建甚?

新的丝氨酸蛋白酶基因ST14(SNC19)在大肠癌中的表达研究

目的 研究新的丝氨酸蛋白酶基因 ST14(SNC19)在大肠癌中的分布和表达情况 ,探讨该基因与淋巴结转移的相关性。方法 采用原位杂交技术检测 30例大肠癌、癌旁黏膜和正常肠黏膜的 ST14表达。结果 30例病人中 ,正常肠黏膜组 ST14表达阳性率 46 .7%,癌旁黏膜组 43.3%,大肠癌组 6 .7%,前两组高于大肠癌组 (P<0 .0 5 )。淋巴结转移组癌旁黏膜 ST14表达阳性率 (6 4.7%)高于无淋巴结转移组 (15 .4%) ,差异有显著性 (P<0 .0 5 )。结论 ST14在部分大肠癌中呈表达缺陷 ,ST14可能作为预测淋巴结转移和判断预后的新指标。

Effect of SNC19/ST14qene overexpression on invasion of colorectal cancer cells

AIM： TO study the effect of SNC19/ST14gene overexpression on invasion in vitro of colorectal cancer cells. METHODS： The adhesion of SlVC19/ST14gene-transfected cells to ECM was measured by MTT assay. The cell movement was evaluated by wound healing assay. Cell invasion and migration were determined by invasion assay in vitro. RESULTS： SNC19/ST14 gene overexpression could enhance invasion of colorectal cancer cells in vitro significantly and influence early cell adherence to ECM, but could not change cell movement significantly. CONCLUSION： SNC19/ST14 gene overexpression increases the local invasion of colorectal cancer cells in vitro. 2005 The WJG Press and Elsevier Inc. All rights reserved.

Expression of serine protease SNC19/matriptase and its inhibitor hepatocyte growth factor activator inhibitor type 1 in normal and malignant tissues of gastrointestinal tract

AIM： To provide the expression profile of serine protease SNC19/matriptase and its inhibitor hepatocyte growth factor activator inhibitor type 1 （HAI-1） in normal and malignant tissues of gastrointestinal tract at mRNA level for further study on their correlations with tumor progression and metastasis. METHODS： Total RNAs were prepared from 37 samples of colorectal cancer tissues, 40 samples of gastric cancer tissues, and their adjacent normal tissues. The expression of SNC19/matriptase and HAI-1 in these samples was detected by real-time fluorescent quantitative PCR using glyceraldehyde-3-phosphate dehydrogenase as internal standard, and the clinical significance for the correlation with clinicopathological parameters was evaluated. RESULTS： In gastric cancer tissues the expression of HAI-1 and SNC19/matriptase was significantly lower than that in the corresponding adjacent normal tissues （Z = -3.280, P= 0.006; Z= -4.651, P= 0.000）. HAI-1：SNC19/matriptase ratio showed no difference between normal and malignant tissues （P〉0.05）. Analysis of clinicopathological parameters showed decreased expression of HAI-1 and HAI-1：SNC19/ matriptase ratio associated with stage Ⅲ/Ⅳ gastric tumors as compared to stage Ⅰ/Ⅱ ones （Z= -2.140, P= 0.031; Z = -2.155, P = 0.031）, and with lymph node-positive gastric cancer tissues as compared to lymph node-negative ones （Z = -2.081, P = 0.036; Z= -2.686, P = 0.006）. The expression of SNC19/matriptase had no relationship with stages and lymph node metastasis （P〉0.05）. The expression of HAI-1 and HAI-1：SNC19/matriptase ratio increased in well-differentiated gastric cancer tissues, but there was no statistical significance （P〉0.05）. The difference of SNC19/matriptase expression was not significant in gastric cancer tissues of different histological differentiation status （P〉0.05）. In colorectal cancer tissues, the expression of HAI-1 and SNC19/matriptase was also markedly lower than that in their adjacent normal tissues （Z= -3.100, P = 0.002; Z= -2.731, P = 0.006）, whereas HAI-1：SNC19/matriptase ratio showed no difference. Decreased expression of HAI-1 was associated with increased invasive depth and lymph node metastasis, but there was no statistical significance （P〉0.05）. The difference of SNC19/matriptase expression and HAI-1： SNC19/matriptase ratio was not significant in different stages and different lymph node metastasis status （P〉0.05）. The expression of SNC19/matriptase, HAI-1 or HAI-1： SNC19/matriptase ratio showed no difference in colorectal cancer tissues of different histological differentiation status （P〉0.05）. CONCLUSION： The expressions of SNC19/matriptase and its inhibitor HAI-1 are decreased in gastrointestinal cancer tissues compared to their normal counterparts, and the decreased expression of HAI-1 may correlate with invasion and lymph node metastasis. The possible mechanisms involved need to be further investigated.