Nakanishi Kiyo, Watanabe Yoshihiko, Maruyama Masato, Yamashita Fumiyoshi, Takakura Yoshinobu, Hashida Mitsuru
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Arch Biochem Biophys. 2002 Jun 15;402(2):201-7. doi: 10.1016/S0003-9861(02)00093-0.
Epithelial cells are an attractive target for local gene delivery in gene therapy for which cytokine genes such as interferon (IFN) genes are promising. However, how the secretion of the gene products is regulated in epithelial cells has been insufficiently investigated. Here, we have studied the secretion polarity of IFN-beta expressed via gene transfection in mouse epithelial Pam-T cells on a bicameral culture system. In transient expression, IFN-beta was predominantly secreted from the cell membrane side on which the transfection was carried out. Meanwhile, the secretion of constitutive IFN-beta from stable transformants was apparently unpolarized. Interestingly, the transformants displayed a polarized secretion of transiently expressed IFN-beta in a transfection-side-dependent manner, their stable IFN-beta secretion remaining unpolarized. These results suggest that epithelial cells have at least dual protein sorting-secretion pathways, transient and stable, for the same secretory proteins, such as IFNs.
上皮细胞是基因治疗中局部基因递送的一个有吸引力的靶点,对于基因治疗而言,诸如干扰素(IFN)基因等细胞因子基因很有前景。然而,上皮细胞中基因产物的分泌是如何调控的,目前尚未得到充分研究。在此,我们利用双室培养系统,研究了在小鼠上皮Pam-T细胞中通过基因转染表达的IFN-β的分泌极性。在瞬时表达中,IFN-β主要从进行转染的细胞膜一侧分泌。同时,稳定转化体中组成型IFN-β的分泌显然是非极化的。有趣的是,转化体以转染侧依赖的方式表现出瞬时表达的IFN-β的极化分泌,其稳定的IFN-β分泌仍是非极化的。这些结果表明,上皮细胞对于相同的分泌蛋白,如IFN,至少具有瞬时和稳定的双重蛋白质分选-分泌途径。