Yang Jie-lin, Zhu Nai-shuo, Wang Ying, Guan Xiao-feng, Zheng Zhao-xin
Lab of Molecular Immunology, State Key Lab of Genetic Engineering, School of Life Sciences, Fudan University.
Mol Biol Rep. 2004 Dec;31(4):241-8. doi: 10.1007/s11033-005-2712-y.
Metastasis and invasion, the important characteristics of malignant tumors, are closely associated with a series of changes in the expression of genes and proteins. In this study, we compare mRNA and protein expression in high and low metastasis adenoid cystic carcinoma cell lines by mRNA suppression subtractive hybridization and two-dimensional electrophoresis combined with peptide mass fingerprint analysis. 34 differentially expressed genes were obtained using suppression subtractive hybridization experiments including 6 highly expressed gene sequences in the high metastasis cell line, and 28 in the low metastasis cell line. RNA dot blot hybridization further confirmed the results after excluding false positives. For protein analysis, ten significantly different protein spots were detected using two-dimensional gel electrophoresis technique combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI- TOF-MS). The results then compare with the SWISS PROT database. These results suggest that high tumor metastasis of adenoid cystic carcinoma is associated with multiple genes whose function include angiogenesis, protein synthesis, signal transduction, modulation of cell cycle, molecular chaperones, and immune co-stimulating molecule. Moreover, the results of the phenotypic function-related expression mapping analysis at the mRNA and protein level revealed obvious complementarities, providing important clues for further study of the molecular mechanism of metastasis, metastasis control and possible targets for cancer gene therapy.
转移和侵袭是恶性肿瘤的重要特征,与基因和蛋白质表达的一系列变化密切相关。在本研究中,我们通过mRNA抑制消减杂交以及二维电泳结合肽质量指纹分析,比较高转移和低转移腺样囊性癌细胞系中的mRNA和蛋白质表达。利用抑制消减杂交实验获得了34个差异表达基因,其中包括高转移细胞系中6个高表达基因序列,以及低转移细胞系中28个。排除假阳性后,RNA斑点杂交进一步证实了结果。对于蛋白质分析,采用二维凝胶电泳技术结合基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOF-MS)检测到10个明显不同的蛋白质斑点。然后将结果与SWISS PROT数据库进行比较。这些结果表明,腺样囊性癌的高肿瘤转移与多个基因相关,其功能包括血管生成、蛋白质合成、信号转导、细胞周期调控、分子伴侣和免疫共刺激分子。此外,在mRNA和蛋白质水平上与表型功能相关的表达图谱分析结果显示出明显的互补性,为进一步研究转移的分子机制、转移控制以及癌症基因治疗的可能靶点提供了重要线索。