含Caspase募集结构域的膜相关鸟苷酸激酶蛋白1与胃肠道淋巴瘤临床病理特征及幽门螺杆菌感染的关系

Relation between Caspase recruitment domain-containing membrane-associated guanylate kinase protein 1, clinicopathological features of gastrointestinal lymphoma also and Helicobacter pylori infection

目的 探讨含Caspase募集结构域的膜相关鸟苷酸激酶蛋白1（CARMA1）表达与胃肠道MALT淋巴瘤和弥漫大B细胞淋巴瘤（DLBCL）临床病理特征及H.pylori感染的关系.方法 选取1999年1月至2011年3月胃肠道DLBCL患者34例和MALT淋巴瘤患者20例,同时选取21例胃肠淋巴组织反应性增生者为对照.用免疫组织化学染色法检测所有研究对象的Ki-67、CARMA1、细胞毒素相关基因A （CagA）蛋白表达情况.用实时定量PCR方法检测28例DLBCL患者和14例MALT淋巴瘤患者手术所获组织标本中CARMA1 mRNA的相对表达量.用硼酸亚甲蓝染色或半套式PCR方法检测病变部位为胃的25例淋巴瘤患者和10例对照者的H.pylori感染情况.计数资料行卡方检验.计量资料行独立样本t检验.采用COX多因素回归分析法进行生存分析.用Kaplan-Meier法绘制生存曲线并行对数秩和检验.结果 28例DLBCL患者的CARMA1 mRNA相对表达量（3.073±1.846）高于14例MALT淋巴瘤患者,差异有统计学意义（F＝ 0.975,P＜0.05）.胃肠道淋巴瘤组的CARMA1蛋白阳性表达率[75.9％（41/54）]高于对照组[47.6％（10/21）],差异有统计学意义（x^2＝5.568,P＜0.05）,其中MALT淋巴瘤组（11/20）低于DLBCL组[88.2％（30/34）],差异有统计学意义（x^2＝5.900,P＜0.05）.在42例接受手术治疗的胃肠道淋巴瘤患者中,高增殖活性者的CARMA1 mRNA相对表达量（2.885±1.837）高于低增殖活性者,临床晚期病例组CARMA1 mRNA相对表达量（4.416±1.0L0）高于临床早期病例组,差异均有统计学意义（F=3.317和2.972,P均＜0.05）.在54例胃肠道淋巴瘤患者中,高增殖活性者的CARMA1蛋白阳性表达率[88.6％（31/35）]高于低增殖活性者（10/19）,差异有统计学意义（x2＝6.847,P＜0.05）.相对11例未感染H.pylori的胃淋巴瘤患者,14例感染H.pylori的胃淋巴瘤患者CARMA1 mRNA相对表达量和CARMA1蛋白表达阳性率差异均无统计学意义（P均＞0.05）.CagA阳性和CagA阴性的H.pylori感染胃淋巴瘤患者CARMA1蛋白表达阳性率分别为11/11和2/3.CARMA1蛋白的表达情况与胃肠道淋巴瘤的预后有关（RR＝ 4.160,P＜0.05）.在获随访的29例胃肠道淋巴瘤患者和18例DLBCL患者中,CARMA1蛋白阳性表达率≥50％者生存状况较＜50％者差,差异均有统计学意义（x^2＝5.383和4.028,P均＜0.05）.结论 CARMA1表达可能参与MALT淋巴瘤和DLBCL的发生、发展,并可能是其不良预后的相关因子;这两种淋巴瘤中CARMA1的表达情况与H.pylori感染无关.

Objective To investigate the relation between the expression of Caspase recruitment domain-containing membrane-associated guanylate kinase protein 1 （CARMA1）,clinicopathological features of gastrointestinal mucosa-associated lymphoid tissue （MALT） lymphoma,diffuse large B cell lymphoma （DLBCL）,and Helicobacter pylori （H.pylori） infection.Methods From January 1999 to March 2011,34 patients with DLBCL and 20 patients with MALT lymphoma were selected,and at same period 21 cases with reactive hyperplasia of gastrointestinal lymphoid tissue were enrolled in as control.The expression of CARMA1,Ki-67 and cytotoxin-associated gene A （CagA） at protein level were examined by immunohistochemistry.The relative expression quantity of CARMA1 mRNA was detected by real-time polymerase chain reaction （PCR）.The condition of H.pylori infection in 25 gastric lymphoma and 10 controls was detected by methylene boric acid and blue staining or semi-nested PCR.Chi-square test was used for counting data analysis,t test for measurement data.Multivariate COX regression analysis was implemented for survival analysis.Survival curve was drawn by Kaplan-Meier method and analyzed by Log-rank test.Results The relative expression quantity of CARMA1 mRNA of 28 patients with DLBLC （3.073±1.846） was higher than that of 14 patients with MALT lymphoma,and the difference was statistically significant （F 0.975,P〈 0.05）.The positive rate of CARMA1 expression at protein level of gastrointestinal lymphoma group （75.9 %,41/54） was higher than that of control group （47.6%,10/21）,and the difference was statistically significant （x2 =5.568,P〈0.05）,and the positive rate of CARMA1 expression of MALT lymphoma group （11/20） was lower than that of DLBCL group （88.2%,30/34）,and the difference was statistically significant （x2 =5.900,P〈0.05）.Among 42 patients with gastrointestinal lymphoma who received surgery,the relative expression quantity of CARMA1 mRNA in cases with high proliferation （2.885±1.837） was higher than that in cases with low proliferation.The expression of CARMA1 mRNA in the cases at advanced stage of the disease （4.416± 1.010） was higher than that in cases at early stage,and the difference was statistically significant （F=3.317 and 2.972,both P〈0.05）.Among 54 patients with gastrointestinal lymphoma,the positive rate of CARMA1 expression at protein level of patients with high proliferation （88.6%,31/35） was higher that that of patients with low proliferation （10/19）,and the difference was statistically significant （x^2 = 6.847,P〈0.05）.There were no significant differences in relative expression quantity of CARMA1 mRNA and the positive rate of CARMA1 expression at protein level between 11 gastric lymphoma patients without H.pylori infection and 14 gastric lymphoma patients with H.pylori infection （both P〉0.05）.The positive rate of CARMA1 expression at protein level of CagA positive and CagA negative H.pylori infected gastric lymphoma patients was 11/11 and 2/3.The expression of CARMA1 at protein level was correlated with the prognosis of gastrointestinal lymphoma （RR=4.160,P〈0.05）.In the 29 cases of patients with gastrointestinal MALT lymphoma and 18 cases of patients with DLBCL who were followed up,the survival situation of gastrointestinal lymphoma patients with CARMA1 positive expression rate over 50% was worse than that of patients with the rate less than 50%,and the difference was statistically significant （x^2=5.383 and 4.028,both P〈0.05）.Conclusions CARMA1 might be involved in the pathogenesis and progression of MALT and DLBCL; and it might be a related factor of poor prognosis.There was no correlation between the expression of CARMA1 and H.pylori infection in these two lymphomas.