Li Zu-guo, Zhao Liang, Liu Li, Ding Yan-qing
Key Laboratory of Molecular Tumor Pathology of Guangdong Province, Department of Pathology, Southern Medical University, Guangzhou, China.
Zhonghua Bing Li Xue Za Zhi. 2007 Jan;36(1):48-52.
To investigate the changes of several protein markers in a metastatic colorectal carcinoma model by serum proteomic analysis.
The pEGFP-N1 plasmid with enhanced expression of green fluorescence protein (EGFP) was transfected into human colon carcinoma cell line SW480 to obtain a stable SW480-EGFP cell line, the SW480-EGFP cells were then injected subcutaneously into nude mice. The harvested tumor cells were implanted orthotopically into the colon of the nude mice. Real-time tumor growth and metastasis formation were visualized by whole-body fluorescent imaging system. Serum samples at different metastatic stages were collected and differential proteomic profiles were investigated by two-dimensional gel electrophoresis (2DE) and matrix-assisted laser absorption/ionization time of flight mass spectrometry (MALDI-TOF MS).
The SW480- EGFP cells enabled to express EGFP stably. The rates of subcutaneous and orthotropic tumor formation were 100%. The metastasis rates to local lymph nodes, liver and lung were 100%, 40% and 30%, respectively. Furthermore, 5 differentially expressed proteins were analyzed by serum proteome technologies, including haptoglobin alpha chain, apolipoprotein E, apolipoprotein A-IV, Ig kappa chain V region chain L and transferrin.
Visualized metastatic model of colorectal carcinoma was successfully established. Several differentially expressed serum proteins collected at different stages after the occurrence of metastasis were identified. These differentially expressed proteins may be candidate serum biomarkers for diagnosis and therapeutic evaluation of colorectal carcinoma metastasis.
通过血清蛋白质组学分析研究转移性结直肠癌模型中几种蛋白质标志物的变化。
将增强型绿色荧光蛋白(EGFP)表达增强的pEGFP-N1质粒转染至人结肠癌细胞系SW480,获得稳定的SW480-EGFP细胞系,然后将SW480-EGFP细胞皮下注射到裸鼠体内。收集收获的肿瘤细胞原位植入裸鼠结肠。通过全身荧光成像系统可视化实时肿瘤生长和转移形成。收集不同转移阶段的血清样本,通过二维凝胶电泳(2DE)和基质辅助激光吸收/电离飞行时间质谱(MALDI-TOF MS)研究差异蛋白质组图谱。
SW480-EGFP细胞能够稳定表达EGFP。皮下和原位肿瘤形成率均为100%。局部淋巴结、肝脏和肺的转移率分别为100%、40%和30%。此外,通过血清蛋白质组技术分析了5种差异表达蛋白,包括触珠蛋白α链、载脂蛋白E、载脂蛋白A-IV、Igκ链V区链L和转铁蛋白。
成功建立了结直肠癌可视化转移模型。鉴定了转移发生后不同阶段收集的几种差异表达的血清蛋白。这些差异表达的蛋白质可能是结直肠癌转移诊断和治疗评估的候选血清生物标志物。