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目的:获得稳定表达小鼠IL-27(mIL-27)基因的小鼠结肠腺癌细胞株(Colon26),研究IL-27基因对小鼠结肠腺癌细胞体内外生物学特性的影响。方法:通过脂质体法将pcDNA3.1(+)-His(B)/mIL-27质粒转染结肠腺癌细胞,筛选建立高表达细胞株。采用Western blot、ELISA、RT-PCR法证实IL-27基因成功转染Colon26。用MTT比色法检测IL-27质粒转染后对Colon26细胞生长的影响;流式细胞术检测IL-27质粒转染后对Colon26细胞凋亡的影响;将Colon26-IL-27细胞接种于BALB/c小鼠的右侧背部皮下,观察其致瘤性。结果:Colon26-IL-27细胞可表达IL-27mRNA并分泌IL-27,IL-27的表达在体外对Colon26细胞的增殖、凋亡均无影响,而在体内则具有明显的抗瘤活性。结论:成功制备Colon26-IL-27细胞株,证明IL-27能在体内明显抑制肿瘤的生长,延长荷瘤小鼠的生存期,具有一定的抗肿瘤作用。
Objective: To obtain mouse colon adenocarcinoma cell line (Colon26) stably expressing mouse IL-27 (mIL-27) gene and study the effect of IL-27 gene on the biological characteristics of mouse colon adenocarcinoma cell in vitro and in vivo. Methods: The pcDNA3.1 (+) - His (B) / mIL-27 plasmid was transfected into colon adenocarcinoma cells by lipofectamine and the highly expressed cell lines were screened. The results of Western blot, ELISA and RT-PCR confirmed that IL-27 gene was successfully transfected into Colon26. The effect of IL-27 plasmid on the growth of Colon26 cells was detected by MTT assay. The effect of IL-27 plasmid on the apoptosis of Colon26 cells was detected by flow cytometry. Colon26-IL-27 cells were inoculated into BALB / c mice on the right back of the skin subcutaneous tumorigenicity was observed. Results: Colon26-IL-27 cells could express IL-27 mRNA and secrete IL-27. The expression of IL-27 had no effect on the proliferation and apoptosis of Colon26 cells in vitro, but had obvious anti-tumor activity in vivo. Conclusion: Colon26-IL-27 cell line was successfully prepared and proved that IL-27 can significantly inhibit the growth of tumor in vivo and prolong the survival of tumor-bearing mice. It has anti-tumor effect.