PDIA1和EF1D在左侧和右侧结肠癌差异表达的实验研究

目的：初步确定蛋白质二硫化异构酶A1和延伸因子1-delta为左侧和右侧结肠癌中差异表达蛋白，为左侧和右侧结肠癌在肿瘤生物学方面的差异提供分子遗传学依据。方法：收集人左侧结肠癌（LSCC）和右侧结肠癌（RSCC）组织标本，进行二维凝胶电泳、质谱分析和生物信息学分离和鉴定左右侧结肠癌中差异表达蛋白质，进一步应用RT—PCR、WesternBlot和免疫组织化学技术检测蛋白质二硫化畀构酶A1和延伸因子1-delta在左侧和右侧结肠癌中的表达状态。结果：筛选并成功鉴定出左侧和右侧结肠癌中16种差异表达蛋白质，与右侧结肠癌比较，lO种蛋白在左侧结肠癌表达上调，6种蛋白在左侧结肠癌表达下调，其中蛋白质二硫化异构酶A1在左侧结肠癌中表达上调，延伸因子1-delta在左侧结肠癌中表达下调，并通过RT-PCR、Western Blot和免疫组化方法证实了在左侧和右侧结肠癌中该两种蛋白表达的差异，与蛋白质组学结果一致。结论：左侧结肠癌和右侧结肠癌的蛋白质组存在差异，其中蛋白质二硫化异构酶A1和延伸因子1—delta在mRNA和蛋白水平均存在差异，这些可能是左侧和右侧结肠癌生物学行为差异的原因。

Experimental Study on PDIA1 and eEF 1 D differential Expression between Left Sided Colon Carcinoma and Right Sided Colon Carcinoma

Objective： To provide molecular genetic basis for oncobiological difference between Left sided colon cancer and Right sided colon cancer by detecting the expression of Left sided colon cancer and Right sided colon cancer including Protein disulfide-isomeraseA 1 and Elongation factor 1. Methods： Tissue samples including Left sided colon cancer and Right sided colon cancer were collect- ed. Protein was separated by using two-dimensional gel electrophoresis （2-DE） and the images of the gels were acquired by the scanner. The peptide mass fingerprintings （PMF） was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and the proteins were identified by data searching with bioinformatics. The expression were assayed by RT-PCR, Western Blot, Immunohistochemical method. Results： 16 protein spots were identified .Compared with right sided colon cancer, 10 pro- teins up-regulated including PDIA1 and 6 proteins down--regulated including EEF-1D in Left sided colon cancer. The expression of PDI- A1 and EEF-1D between right sided colon cancer and left sided colon cancer were confirmed by RT-PCR, Western Blot and Immunohistochemical method. Conclusions： There were differentially expressed proteins between Left sided colon cancer and Right sided colon cancer, especially for PDIA 1 and EEF- 1 D, which were cause by oncobiological difference between Left sided colon cancer and Right sided colon cancer.