ILK、Hsp90相互作用及在胃癌中表达的研究

目的:探讨ILK与Hsp90在胃癌组织中的相互作用,了解它们及相关因子E-cadherin与胃癌临床病理的关系,以及胃癌患者血清中手术前后ILK与Hsp90含量的变化.方法:用免疫共沉淀,蛋白杂交的方法研究ILK与Hsp90的直接相互作用,用免疫组织化学方法测定ILK,Hsp90,E-cadherin和MMP2在胃癌及癌旁组织表达,比较它们与胃癌临床病理的关系,使用Elisa方法,测定ILK与Hsp90在胃癌患者血清中手术前后含量的变化.结果:用ILK抗体免疫沉淀下来的蛋白中检测有Hsp90蛋白存在.ILK在40例胃癌组织中有28例表达阳性,阳性率为70%,而Hsp90则在37例有高表达,阳性率为.92.5%.其中28例ILK阳性者中有27位Hsp90表达阳性.高分化胃癌分别有50%和75%表达,而低分化者为75%和100%.E-cadherin在正常胃粘膜上皮细胞均为阳性表达,而只有40%的胃癌组织呈阳性.其中高分化者有75%阳性表达,而低分化者仅为32.1%,没有转移的为90%,有转移的仅是23.3%.MMP2在癌组织中的表达阳性率为75%,高于癌旁正常组织,高分化者有50%阳性表达,而低分化者为78.6%,没有转移的为50%,有转移的是83.3%.用Elisa方法测定血清中ILK,Hsp90抗体的含量.结果表明,手术前ILK、Hsp90的含量分别为171.0ng/ml和199.0ng/ml,而手术后分别下降为142.6ng/ml和161.6ng/ml.结论:实验结果证明胃癌组织Hsp90与ILK的相互作用是直接的结合.ILK,Hsp90,E-cadherin和MMP2的表达与肿瘤分化程度显著相关,与淋巴结转移、浸润深度、TNM分期也有一定的关系.这些因子的表达对胃癌的分化程度有重要作用,血清的检测ILK和Hsp90可能对术后状况的监控有一定指导意义.

The Interaction of ILK, Hsp90 and the Expression in Gastric Carcinoma

Objective: To study the interaction between ILK and Hsp90 in gastric carcinoma, and find the relation between the genes and related factors, such as E-cadherein and MMP2, and the clinical pathology of gastric carcinoma, and the change between the content of ILK and of Hsp90 in the serum of patients with gastric cancer, before and after the surgical operation. Methods: The interaction between ILK and Hsp90 in gastric carcinoma using the methods of coimmunoprecipitate and western was investigated. Immunohisto-chemistry was applied to detect the expression of ILK, Hsp90, MMP2, E-cadherin in 40 gastric cancer. The content of ILK and Hsp90 was tested in patient's serium with gastric carcinoma before and after surgical operation. Results: The Hsp90 protein was studied by coimmunoprecipitating with ILK antibody. There was the evidence of interaction between ILK and Hsp90. Strong expressions of ILK protein was detected in 28/40(70%) of primary gastric carcinoma, but there was a high express of Hsp90 in 37 of the samples and the rate was 92.5%. There was the positive expression of Hsp90 in 27 of the 28 ILK positive samples. The expression of ILK and Hsp90 in well-differentiated gastric cancer accounted for 50% and 75% respectively, while in low-differentiation type, it accounted for 75% and 100%. Positive expression of E-Cadherin occurred in all the normal stomach samples, but only 40% of the expression in gastric carcinoma was positive, among which 75% positive expression occurred in the well-differentiated cancer, 32.1% in low-differentiated cancer and 90% of the cases have no nodal metastasis, and the nodal metastasis occurred in 23.3% of the cases. The positive expression of MMP2 in gastric carcinoma was 75%, and it was higher than that in the normal paraneoplastic tissue. Elisa method was used to detect the content in the antibody of ILK and Hsp90 in serum of the patients, before and after surgical operation. The results showed that the content of ILK and Hsp90 was 171.0 mg/ml and 199.0 mg/ml before operation and after operation, the content of the Hsp90 was 142.6 ng/ml and 161.6 ng/ml, respectively. Conclusion: The results show that the interactions between Hsp90 and ILK is a direct combination. The strong expression of ILK, Hsp90 and MMP2 exhibits significantly the association with low differentiation type and deep invasion of gastric carcinoma. The expression of these factors play an important role on the differentiation of gastric carcinoma, and detection on the content of ILK and Hsp90 in serum of the patients may has a definite guidance for monitoring on postoperative condition of the patients.