【作者】 张敬坡；

【导师】 李勇； 宋振川；

【作者基本信息】 河北医科大学 ， 外科学， 2008， 硕士

【摘要】 目的:胃癌是消化道最常见的恶性肿瘤之一,具有发病率高、死亡率高、预后差等特点,虽然目前对胃癌采取了多手段的综合治疗,且生存率较以往有所提高,但仍无突破性进展。侵袭和转移是影响肿瘤患者治疗效果和预后的重要因素,控制肿瘤细胞的侵袭和转移仍是当今肿瘤研究中的重要课题。细胞间粘附分子-l(ICAM-l)是一种非常重要的细胞粘附分子,它可以调节细胞与细胞、细胞与细胞外基质间的相互结合,被认为在肿瘤的局部浸润和远处转移中起着重要的作用。ICAM-l能辅助稳定T细胞受体介导的抗原呈递细胞(APC)和淋巴细胞的结合,是重要的细胞免疫协同激活因子。粘着斑激酶(FAK)是一种非受体蛋白酪氨酸激酶,分布在细胞粘着斑部位。近年来研究表明,在多种肿瘤组织中都有FAK蛋白表达的显著增加,且FAK蛋白的表达与肿瘤的发生及生物学行为有一定的相关性。本研究通过检测ICAM-1、LFA-1及FAK在胃癌组织、正常胃粘膜及淋巴结中的表达情况及T细胞亚群和NK细胞的表达情况,研究ICAM-1、LFA-1及FAK对胃癌发生和发展的影响及其与胃癌临床生物学行为之间的内在联系,为进一步探讨肿瘤免疫耐受的机制及改进肿瘤的免疫治疗和基因治疗的方法提供更多的理论依据。方法:选取河北医科大学第四医院外三科2007年7月~2007年11月胃癌手术切除标本共60例。采集胃癌组织标本、正常胃粘膜(距肿瘤边缘>5.0cm)及淋巴结。采用流式细胞仪测定癌组织和外周血中T淋巴细胞亚群和NK细胞的活性。用免疫组织化学方法(S-P法)检测胃癌组织、正常胃粘膜及淋巴结中ICAM-1、LFA-1及FAK蛋白表达情况。结果:1胃癌患者外周血及癌组织中T淋巴细胞亚群及NK分布情况胃癌组织中CD4+、CD8~+、NK细胞计数和CD4/CD8比值分别为37.44±8.90、36.40±8.86、9.28±3.37和0.51±0.11,外周血中的含量分别为52.01±9.55、20.66±5.19、19.94±6.28、1.57±0.49;外周血中CD4+、NK细胞计数和CD4+/CD8~+比值均显著高于胃癌组织(P<0.05),CD8~+细胞反之(P<0.05)。2胃癌组织中ICAM-1、LFA-1及FAK的表达与其临床生物学行为的关系2.1胃癌组织中ICAM-1的表达与其临床生物学行为的关系ICAM-1在胃癌组织的阳性表达率为53.33%,低于正常胃粘膜中的阳性表达率76.67%,二者比较差异有显著性(P<0.05);在分化程度中低分化组的阳性表达率为43.59%,低于高/中分化组的阳性表达率71.43%,二者比较差异有显著性(P<0.05);在有淋巴结转移组的阳性表达率为43.90%,低于无淋巴结转移组的阳性表达率73.68%,二者比较差异有显著性(P<0.05);在临床分期中III-IV期组的阳性表达率为45.46%,低于I-II期组的阳性表达率75.00%,二者比较差异有显著性(P<0.05);ICAM-1表达与患者性别、年龄、肿瘤部位、肿瘤大小、侵润深度及Borrmann分型均无关(P>0.05)。2.2胃癌组织中LFA-1的表达与其临床生物学行为的关系LFA-1在胃癌组织中的阳性表达率为43.33%,低于正常胃粘膜中的阳性表达率61.67%,二者比较差异有显著性(P<0.05)。低分化组的阳性表达率为33.33%,低于高/中分化组的阳性表达率61.90%,二者比较差异有显著性(P<0.05);在有淋巴结转移组的阳性表达率为29.27%,低于无淋巴结转移组的阳性表达率73.68%,二者比较差异有显著性(P<0.05);在临床分期中III-IV期组的阳性表达率为31.82%,低于I-II期组的阳性表达率75.00%,二者比较差异有显著性(P<0.05);LFA-1表达与患者性别、年龄、肿瘤部位、肿瘤大小、侵润深度及Borrmann分型均无关(P>0.05)。2.3胃癌组织中FAK的表达与其临床生物学行为的关系FAK在胃癌组织中的阳性表达率为70.00%,高于正常胃粘膜中的阳性表达率5.00%,二者比较差异有显著性(P<0.05);低分化组的阳性表达率为79.49%,高于高/中分化组的阳性表达率52.38%,二者比较差异有显著性(P<0.05);在浸润深度中T3+T4组的阳性表达率为77.08%,高于T1+T2组的阳性表达率41.67%,二者比较差异有显著性(P<0.05);在临床分期中III-IV期组的阳性表达率为79.55%,高于I-II期组的阳性表达率43.75%,二者比较差异有显著性(P<0.05);在有淋巴结转移组的阳性表达率为80.49%,高于无淋巴结转移组的阳性表达率47.37%,二者比较有显著性(P<0.05);FAK表达与患者性别、年龄、肿瘤部位、肿瘤大小及Borrmann分型均无关(P>0.05)。3胃癌区域淋巴结中ICAM-1、LFA-1及FAK的表达情况胃癌区域淋巴结中,有转移淋巴结中FAK的阳性表达率为90.63%,高于无转移淋巴结中的阳性表达率35.71%,二者比较差异有显著性(P<0.05);有转移淋巴结中ICAM-1的阳性表达率为40.63%,无转移淋巴结中的阳性表达率50.00%,二者比较无显著性差异(P>0.05);有转移淋巴结中LFA-1的阳性表达率为34.38%,无转移淋巴结中的阳性表达率42.86%,二者比较无显著性差异(P>0.05)。4胃癌组织中ICAM-1、LFA-1与CD4~+/CD8~+、NK细胞表达的相关性分析癌组织中ICAM-1与CD4~+/CD8~+、NK细胞的表达呈正相关(rs=0.900、rs=0.815,P<0.05);LFA-1与CD4~+/CD8~+、NK细胞的表达呈正相关(rs=0.954、rs=0.787,P<0.05)。5胃癌组织中ICAM-1、LFA-1及FAK表达的相关性分析胃癌组织中ICAM-1与FAK的表达呈明显负相关(rs= -0.387,P<0.01)。LFA-1与FAK的表达无明显相关(rs= -0.240,>0.05)。ICAM-1与LFA-1的表达无明显相关(rs=0.097,P>0.05)。结论:1胃癌患者癌组织中的CD4~+、NK细胞计数和CD4~+/CD8~+比值较外周血中的低,而CD8~+显著增高,提示胃癌患者癌组织局部呈现一个免疫低下或抑制状态。2胃癌组织中ICAM-1、LFA-1表达降低及FAK表达增高可能是胃癌发生的早期事件。三者表达失衡均与胃癌的分化程度,淋巴结转移,临床病理分期相关,在一定程度上反映了胃癌的进展过程,可作为监测胃癌进程的重要指标。3胃癌组织中ICAM-1及LFA-1蛋白的表达率明显低于正常胃组织,并随着肿瘤恶性程度的增高而降低,且NK细胞、CD4~+/CD~+8表达与两者表达呈正相关,提示ICAM-1/ LFA-1对T淋巴细胞第二信使的刺激减少,导致T细胞增殖与活化功能不足,不能产生足够的抗肿瘤免疫反应,从而导致胃癌细胞免疫逃逸现象。4胃癌组织中FAK与ICAM-1的表达呈明显负相关,说明在胃癌的发生发展过程中FAK抑制了ICAM-1的表达,在一定程度上抑制了机体的免疫功能,促进了胃癌细胞的免疫逃逸作用。更多还原

【Abstract】 Objective:Gastric cancer is the most common malignant tumor of gastrointestinal tract, with high incidence rate, high death rate and bad prognosis. There is no any significant break through in its treatment, although the gastric cancer has been adopted multi-method therapies and its survival rate has risen. Invasion and metastasis is the most important factor to influence tumor patient’s therapeutic effieacy and prognoses, to control tumor cell’s invasion and metastasis is still an important topic in today’s tumor research.Intercellular adhesion molecule-1(ICAM-1) can regulate the adhesion between cells or the cell and extracellular matrix and play important roles in the infiltration and metastasis of the tumor. ICAM-1 is an accessory molecule stabilizing the T-cell receptor-mediated binding between antigen-presenting cells and T lymphocytes,and serves as a cofactor in activation of the cellular immune response.FAK is a non-receptor protein tyrosine kinase in cell adhesion lesion. Recent studies have shown that in a variety of tumor tissue have significantly increased expression of FAK, also FAK expression and the biological behavior and the occurrence of certain relevance.This study was to detect the expression of ICAM-1, LFA-1, FAK, T lymphocyte subset and NK cell in gastric carcinoma tissues, regional lymph node tissues and normal gastric mucosa. To investigate the relationships between the expression of ICAM-1, LFA-1 and FAK and clinical pathological characteris- tics of gastric cancer, and reveal the possible mechanisms of tumor’s escape from the body surveillance. In order to provide more theoretical evidence for the improvements of immuno- therapy and gene therapy for tumor.Methods:In this study, we selected 60 samples of gastric carcinoma, normal gastric mucosa(distance from the edge of tumor >5.0cm)and regional lymph node separately from 60 patients. All patients had gastrectomy in the 4th hospital of Hebei Medical University from July 2007 to November 2007. In this study, we detected the marks of T lymphocyte subsets and NK cells in tumorous tissues and peripheral blood from patients of gastric cancer with Flow Cytometrye. we used S-P immunohistochemical technique to detect the expressions of ICAM-1、LFA-1 and protein FAK in gastric carcinoma tissues, normal gastric mucosa and regional lymph node.Results:1 Distribution of T cell subgroup and NK cell in gastric cancer and peripheral bloodThe ratio of CD4~+, CD8~+, NK cell and CD4~+/CD8~+ in gastric cancer tissue were 37.44±8.90, 36.40±8.86, 9.28±3.37, 0.51±0.11 respectively; the ratio of CD4~+, CD8~+, NK cell and CD4~+/CD8~+ in peripheral blood were 52.01±9.55, 20.66±5.19, 19.94±6.28, 1.57±0.49 respectively. Their ratio of CD4~+, NK cell and CD4~+/CD8~+ in gastric cancer were lower than them in peripheral blood respectively, the differences were statistically significant(P<0.05). The ratio of CD8~+ T cell were higher in gastric cancer than it in peripheral blood, the differences were statistically significant (P<0.05).2 Correlations between the expression of ICAM-1, LFA-1 and FAK in gastric cancer tissues and clinical biological characteristics2.1 Correlations of ICAM-1 expression and clinicopathologic parameters of gastric cancerThe positive rates of ICAM-1 were 53.33% in tissues of gastric carcinoma, which were lower than the rates of 76.67% in normal gastric mucosa, the comparison of the two shows significant difference (P<0.05). The positive rates in poorly differentiated group(43.59%) were significantly lower than well/moderately differentiated group(71.43%; P<0.05) in the degree of differentiation. The positive rates of the group with lymph node metastasis (43.90%) were significantly lower than that without lymph node metastasis (73.68%; P<0.05). In clinical stages, the positive rates in the group of III-IV stage (45.46%) were significantly lower than group of I-II stage (75.00%; P<0.05). The expression of ICAM-1 hadn’t relation with the age, gender, borrmann type, depth of infiltration, and tumor general typing ( P>0.05).2.2 Correlations of LFA-1 expression and clinicopathologic parameters of gastric cancerThe positive rates of LFA-1 were 43.33% in tissues of gastric carcinoma, which were lower than the rates of 61.67% in normal gastric mucosa, the comparison of the two shows significant difference (P<0.05). The positive rates in poorly differentiated group (33.33%) were significantly lower than well/moderately-differentiated group (61.90%; P<0.05) in the degree of differentiation. The positive rates of the group with lymph node metastasis (29.27%) were significantly lower than that without lymph node metastasis (73.68%; P<0.05). In clinical stages, the positive rates in the group of III-IV stage (31.82%) were significantly lower than group of I-II stage (75.00%; P<0.05). The expression of LFA-1 hadn’t relation with the age, gender, borrmann type, depth of infiltration, and tumor general typing (P>0.05).2.3 Correlations of FAK expression and clinicopathologic parameters of gastric cancerThe positive rates of FAK were 70.00% in tissues of gastric carcinoma, which were higher than the rates of 5.00% in normal gastric mucosa, the comparison of the two shows significant difference (P<0.05). The positive rates in poorly differentiated group (79.49%) were significantly higher than well/moderately differentiated group (52.38%, P<0.05) in the degree of differentiation. In the degree of infiltration, the positive rates in the group of T3+T4 (77.08%) were significantly higher than the rate in T1+T2 (41.67%; P<0.05). In clinical stages, the positive rates in the group of III-IV stage (79.55%) were significantly higher than group of I-II stage (43.75%; P<0.05). The positive rates of the group with lymph node metastasis (80.49%) were significantly higher than that without lymph node metastasis (47.37%; P<0.05). The expression of FAK hadn’t relation with the age, gender, borrmann type, and tumor general typing (P>0.05).3 Expression of ICAM-1, LFA-1 and FAK in gastric regional lymph node tissuesIn gastric regional lymph node tissues, the expressions of FAK in the group with lymph node metastasis (90.63%) were higher than that without lymph node metastasis (35.71%), the comparison of the two shows significant difference (P<0.05). the expressions of ICAM-1 in the group with lymph node metastasis (40.63%) were lower than that without lymph node metastasis (50.00%), the comparison of the two shows no significant difference (P>0.05); the expressions of LFA-1 in the group with lymph node metastasis (34.38%) were lower than that without lymph node metastasis (42.86%), the comparison of the two shows no significant difference (P>0.05).4 Correlations of the expressions of ICAM-1, LFA-1and the T cell subgroup, NK cell in the gastric cancer tissueThe expression of ICAM-1 in gastric cancer tissue were significantly and positive correlated with the ratio of CD4+/CD8~+ and NK cell(rs=0.900、rs=0.815,P<0.05). The expression of LFA-1 in gastric cancer tissue were significantly and positive correlated with the ratio of CD4+/CD8~+ and NK cell(rs=0.954、rs=0.787,P<0.05)5 Correlations of the expressions of ICAM-1, LFA-1 and FAK in the gastric cancer tissueA significant negative correlation was observed between the expression of ICAM-1 and FAK in gastric carcinoma tissues (rs = -0.387, P<0.01). The expression of LFA-1 hadn’t relation with the ICAM-1 and FAK in gastric carcinoma tissues (P>0.05).Conclusion:1 The local immune function in gastric cancer patient was lower than that in peripheral blood. The rate of CD4+、NK cell and CD4+/CD8~+ in gastric cancer tissue were higher than those in the peripheral blood, but the rate of CD8~+ T cell was higher, the local immune function was low or suppressed.2 The expressions of ICAM-1, LFA-1, FAK may be involved into the occurrence and progression of gastric carcinoma. The abnormal expressions of FAK, ICAM-1 and LFA-1 may be one of the earlier of gastric carcinoma. The abnormal expressions of FAK, ICAM-1 and LFA-1 correlates with degree of infiltration, lymph node metastasis and TNM stages of gastric carcinoma, and it reflects the progression of gastric carcinoma in certain degree. 3 The expression of ICAM-1 and LFA-1 in gastric cancer tissue were significantly and positively correlated with NK cell and CD4~+/CD8, indicating that ICAM-1/LFA-1 was significan- tly correlated with the immune function in the genesis and development of gastric cancer. Gastric cancer can decrease cancer tissue ICAM-1/LFA-1 expression to escape body immune system through some mechanism during its occurring and developing. An important cause of the escape may be the failure of the second messenger stimulus to T-lymphocyte from gastric carcinoma cells.4 In gastric carcinoma tissues, the expressions of FAK and ICAM-1 have negative correlation. It shows that the expressions of FAK has inhibited the expressions of ICAM-1, which inhibits the immune function of body in a certain degree, facilitates the immune escape of gastric carcinoma cells.更多还原