Objective: To explore role of MIER3 gene in the development and progression of human colorectal carcinoma (CRC) and analyze the proteins that interact with MIER3 using bioinformatic techniques.
Methods: MIER3 mRNA and protein expressions were detected in 8 CRC biopsy samples and paired adjacent tissues using real-time PCR and Western blotting. A recombinant eukaryotic expression vector pcDNA3-MIER3 was constructed and its effect on the proliferation and invasion of CRC cells were tested using CCK8 assay and Transwell migration assay. Bioinformatic methods were used to predict and analyze MIER3-interacting proteins.
Results: MIER3 was obviously down-regulated in the 8 CRC tissues as compared with the paired adjacent tissues. In human CRC cell line DLD1, MIER3 overexpression induced by transfection of the cells with pcDNA3-MIER3 significantly inhibited the cell proliferation and suppressed cell invasiveness in vitro. Bioinformatics analyses indicated that NAT9 was a potential MIER3-interacting protein and MIER3 was probably associated with tumor susceptibility.
Conclusion: MIER3, which is obviously down-regulated in CRC tissues, is closely associated with the proliferation and invasion of CRC, and NAT9 protein is a probable MIER3-interacting protein.
目的: 构建基因真核表达载体并探讨MIER3对结直肠癌细胞生物学特性的影响，应用生物信息学方法对MIER3及其相互作用蛋白进行预测分析。
方法: 应用Real-time PCR及Western blot方法检测MIER3在8例结肠癌及相应癌旁正常组织中mRNA和蛋白表达水平的差异表达；通过构建真核表达载体pcDNA3-MIER3，应用CCK8增殖实验、Transwell侵袭实验检测过表达MIER3对结直肠癌细胞株生物学特性的影响，进一步应用生物信息学方法对MIER3及其相互作用蛋白进行预测和分析。
结果: MIER3在8例结肠癌组织中其mRNA及蛋白水平的表达明显低于相应正常癌旁组织；酶切及测序结果显示成功克隆构建pcDNA3-MIER3真核表达载体；CCK8增殖实验、Transwell侵袭实验结果显示过表达MIER3可明显抑制结直肠癌细胞株的增殖及侵袭能力；应用模式生物果蝇相互作用蛋白数据库DIP和基于保守的蛋白相互作用分析方法，预测NAT9可能是MIER3的潜在相互作用蛋白，提示MIER3可能与肿瘤的易感性相关。
结论: MIER3与结直肠癌增殖、侵袭密切相关，为进一步深入阐明MIER3在结直肠癌发生发展中的作用奠定基础