肺巨细胞癌高低转移株转移能力差异相关分子的研究  

Study on differentially expressed molecules influencing the metastatic potential between highly and poorly metastatic human lung giant cell carcinoma

作　　者：蒋代凤[1] 陆应麟[2] 邱宗荫[3] 贺福初[1] 

机构地区：[1]军事医学科学院放射医学研究所 [2]军事医学科学院基础医学研究所,北京100850 [3]重庆医科大学

出　　处：《中华肿瘤杂志》2003年第6期531-534,共4页Chinese Journal of Oncology

基　　金：国家重点基础研究项目(G2 0 0 1CB51 0 2 0 1 ;2 0 0 2CB51 31 0 5);国家自然科学基金资助项目( 39730 2 70 ;3990 6 0 0 )

摘　　要：目的　鉴定在肺巨细胞癌高、低转移株差异表达的转移相关分子 ,探讨肺巨细胞癌的转移机制。方法　以肺巨细胞癌高、低转移株 (即PLA80 1D、PLA80 1C)为转移模型 ,通过细胞体外迁移和侵袭实验 ,验证高、低转移株间细胞体外迁移和侵袭能力的差异。以明胶酶谱实验 ,分析肿瘤细胞分泌基质金属蛋白酶 (MMP 2和MMP 9)的活性 ;以Westernblot技术 ,检测肿瘤细胞分泌的MMP 2、MMP 9、组织基质金属蛋白酶抑制剂 (TIMP 1和TIMP 2 )的蛋白水平 ,以及p53、细胞增殖核抗原 (PCNA)、p16、CD44(V6)异构体、细胞骨架蛋白 (CK18)、上皮细胞间黏附分子 (E cadherin)和肿瘤转移抑制蛋白(nm2 3 H1)的蛋白水平 ;以半定量RT PCR技术 ,检测细胞内MMP 2、MMP 9、TIMP 1、TIMP 2和血管内皮生长因子 (VEGF)的mRNA水平。结果　肺巨细胞癌高转移株PLA80 1D的细胞体外侵袭能力显著高于低转移株PLA80 1C ,大约为后者的 4倍。PLA80 1D细胞株分泌的MMP 2活性高于PLA80 1C细胞株 ,其原因主要是由于分泌的MMP 2蛋白以及细胞内MMP 2mRNA水平显著增高所致。在PLA80 1D细胞株内 ,p53、PCNA、CK18蛋白和VEGFmRNA为显著高表达 ,p16、E cadherin和nm2 3 H1蛋白为显著低表达 ,而MMP 9、TIMP 1、TIMP 2和CD44(V6)蛋白的表达在高。Objective To study the metastasis-associated molecules differenti ally expressed in highly and poorly metastatic sublines and the mechanism of met astasis in lung giant cell carcinoma. MethodsHighly and poorly metastatic su blines (PLA801D and PLA801C)were used as metastasis model. Cell motility and i nvasion assay in vitro were first compared between the two sublines. Then, gelat in zymography analysis was used to determine the MMP-2 and MMP-9 activity. The p rotein expression level of secreted MMP-2, MMP-9, TIMP-1, TIMP-2 and intrace llul ar expression level of p53, p16, PCNA, CD44(V6) isomeride, E-cadherin, CK18, n m23-H1 as well as the mRNA expression level of MMP-2, MMP-9, TIMP-1, TIMP-2 , VEG F were compared through Western blot. Semi-quantitative RT-PCR analysis was used to determine the intracellular mRNA expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and VEGF. Results The in vitro cell invasion potential of highly metastatic su bline PLA801D was significantly higher than that of poorly metastatic subline PL A801C by about 4 folds, while the cell motility potential was similar. The secre ted MMP-2 activity was notably higher in PLA801D, which was initiated by the hi g her expression of MMP-2 at protein and mRNA level. In addition, the expression l evel of p53?PCNA?CK18 protein and VEGF mRNA were significantly higher, while t he expression level of p16?E-cadherin and nm23-H1 protein were significantly lo wer in PLA801D. Some molecules such as MMP-9,TIMP-1,TIMP-2,CD44(V6)isom eri de, which had been reported to be associated with tumor metastasis, were not obs erved to change significantly between the two sublines. Conclusion There are significant differences in metastatic potential and phenotypes between highly and poorly metastatic sublines of lung giant cell carcinoma. Some differ entially expressed molecules might be playing roles in promoting or inhibiting m etastasis of lung giant cell carcinoma, which may be useful to elucidate the mec hanism of metastasis.

关 键 词：肺肿瘤 巨细胞癌 肿瘤转移 转移机制 

分 类 号：R734.2[医药卫生—肿瘤]