胃癌组织和转移淋巴结中丝氨酸蛋白酶抑制剂B1表达与肿瘤侵袭转移的关系

Expression of Serpin B1 in gastric cancer,metastatic lymph nodes tissues and its relationship with invasion,metastasis of gastric cancer

目的检测胃癌组织和转移淋巴结中丝氨酸蛋白酶抑制剂B1(Serpin B1)的蛋白质表达情况,探讨Serpin B1与肿瘤侵袭转移的关系。方法采用免疫组织化学方法检测80例胃癌组织、区域淋巴结和癌旁正常胃黏膜石蜡标本中的Serpin B1、周期素D1(cyclin D1)、细胞间黏附因子-1(ICAM-1)、基质金属蛋白酶-9(MMP-9)的蛋白质表达情况;采用Western印迹法检测8例胃癌患者的胃癌组织、区域淋巴结和癌旁正常胃黏膜新鲜标本中Serpin B1蛋白质的表达情况。收集患者的临床资料,记录其临床特征,分析胃癌组织中Serpin B1蛋白质表达与临床特征的关系。测定并比较原发肿瘤、阳性转移淋巴结组织、癌旁正常组织中Serpin B1、cyclin D1、ICAM-1、MMP-9蛋白质表达水平。结果浸润深度较深、临床分期较晚、分化程度差、淋巴结转移阳性和有脉管瘤栓的胃癌患者中Serpin B1蛋白质表达阳性患者的比例显著高于浸润深度较浅、临床分期较早、分化程度好、淋巴结转移阴性和无脉管瘤栓的胃癌患者(P值均<0.05)。胃癌组织和转移淋巴结组织中Serpin B1、cyclin D1、ICAM-1、MMP-9蛋白质表达阳性率均显著高于癌旁正常组织(P值均<0.01);转移淋巴结中Serpin B1、MMP-9蛋白质表达阳性率显著高于胃癌组织(P值分别<0.01、0.05),cyclin D1、ICAM-1蛋白质表达阳性率与胃癌组织间的差异均无统计学意义(P值均>0.05)。Western印迹法检测结果显示,转移淋巴结组织中Serpin B1蛋白质水平显著高于胃癌组织(t=2.791,P=0.014)和癌旁正常组织(t=11.828,P<0.001),胃癌组织中Serpin B1蛋白质水平显著高于癌旁正常组织(t=7.247,P<0.001)。Spearman相关分析显示,胃癌组织与转移淋巴结中Serpin B1蛋白质(r=0.465 2)、cyclin D1蛋白质(r=0.352 2)表达呈正相关(P值均<0.01),与ICAM-1蛋白质(r=0.208 6)、MMP-9蛋白质(r=0.145 8)表达不相关(P值均>0.05)。胃癌组织中Serpin B1蛋白质表达与ICAM-1蛋白质(r=0.342 2)、MMP-9蛋白质(r=0.326 8)表达呈正相关(P值均<0.01),与cyclin D1蛋白质表达不相关(r=0.164 6,P=0.144 6);转移淋巴结中Serpin B1蛋白质表达与MMP-9蛋白质表达呈正相关(r=0.305 2,P=0.018 0)。结论胃癌组织中Serpin B1蛋白质的表达存在异常,可能通过调节ICAM-1、MMP-9蛋白质参与胃癌的侵袭转移。

Objective To detect the expression of serine protease inhibitor B1 （Serpin B1） in gastric cancer and metastatic lymph node tissues, and to explore the relationship between Serpin B1 and invasion, metastasis of gastric cancer. Methods Immunohistochemical staining was applied to test protein expression of Serpin B1, cyclin D1, intercellular adhesion molecule-1 （ICAM-1）, and matrix metalloproteinase 9 （MMP-9） in 80 cases of human gastric cancer tissues, regional lymph node tissues and adjacent gastric mucosal tissues. Fresh specimens of gastric cancer tissues, regional lymph node tissues and adjacent gastric mucosal tissues from another 8 patients with gastric cancer were collected and treated with Western blot assay to test the expression of Serpin B1 protein. Clinical data and clinicopathological features were collected. The correlation between protein expression of Serpin B1 in gastric cancer tissues and clinical features were analyzed. Results The protein expression levels of Serpin B1 protein in gastric carcinoma tissues were positively correlated with tumor depth of invasion, clinical staging, differentiation degree, lymphatic metastasis, and vessel invasion （all P〈0.05）. The expressions of Serpin B1, cyclin D1, ICAM-1 and MMP-9 proteins in gastric cancer tissues and metastatic lymph node tissues were significantly stronger than those in the adjacent gastric tissues （all P〈0. 01）. The expressions of Serpin B1 and MMP-9 proteins in metastatic lymph node tissues were significantly stronger than those in gastric cancer tissues （P〈0.01, 0.05）, whereas there was no statistical difference in the protein expression of cyclin D1 or ICAM-1 between metastatic lymph node tissues and gastric cancer tissues （both P〉0. 05）. The results of Western blot showed that the protein expression level of Serpin B1 in metastatic lymph node tissues was significantly higher than that in gastric cancer tissues （ t = 2. 791, P = 0. 014） and adjacent gastric tissues （ t = 11. 828, P〈0. 001）; and that the protein expression level of Serpin B1 in gastric cancer tissues was significantly higher than that in adjacent gastric tissues （ t = 7. 247, P 〈0. 001 ）. Spearman correlation analysis showed that Serpin B1 and cyclin D1 proteins were positively correlated in metastatic lymph nodes tissues and gastric cancer tissues （r = 0. 465 2, r = 0. 352 2, both P〈O. 01）. In gastric cancer tissues, Serpin B1 protein expression was positively correlated with lOAM-1 （ r = 0. 342 2） and MMP-9 （ r = 0. 326 8, both P〈0. 01 ）, but not correlated with cyclin D1 （r = 0. 164 6, P = 0. 144 6）. In metastatic lymph nodes, positive correlation was found between Serpin B1 and MMP-9 proteins （r=0.305 2, P=0.018 0）. Conclusion Serpin B1 protein is abnormally expressed in gastric cancer tissues, and it may be involved in the invasion and metastasis of gastric cancer by interacting with ICAM-1 and MMP-9.