Yan Fei, Ruan Xu-zhi, Yang Han-shuo, Yao Shao-hua, Zhao Xin-yu, Gou Lan-tu, Ma Fan-xin, Yuan Zhu, Deng Hong-xin, Wei Yu-quan
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
J Biomed Biotechnol. 2010;2010:134764. doi: 10.1155/2010/134764. Epub 2010 May 4.
Apoptosis plays an important role in embryonic development. PNAS-4 has been demonstrated to induce apoptosis in several cancer cells. In this study, we cloned Xenopus laevis PNAS-4 (xPNAS-4), which is homologous to the human PNAS-4 gene. Bioinformatics analysis for PNAS-4 indicated that xPNAS-4 shared 87.6% identity with human PNAS-4 and 85.5% with mouse PNAS-4. The phylogenetic tree of PNAS-4 protein was also summarized. An analysis of cellular localization using an EGFP-fused protein demonstrated that xPNAS-4 was localized in the perinuclear region of the cytoplasm. RT-PCR analysis revealed that xPNAS-4, as a maternally expressed gene, was present in all stages of early embryo development. Whole-mount in situ hybridization showed that xPNAS-4 was mainly expressed in ectoderm and mesoderm. Furthermore, microinjection of xPNAS-4 mRNA in vivo caused developmental defects manifesting as a small eye phenotype in the Xenopous embryos, and as a small eye or one-eye phenotype in developing zebrafish embryos. In addition, embryos microinjected with xPNAS-4 antisense morpholino oligonucleotides (MOs) exhibited a failure of head development and shortened axis.
细胞凋亡在胚胎发育中起重要作用。PNAS - 4已被证明可诱导多种癌细胞发生凋亡。在本研究中，我们克隆了非洲爪蟾的PNAS - 4（xPNAS - 4），它与人PNAS - 4基因同源。对PNAS - 4的生物信息学分析表明，xPNAS - 4与人类PNAS - 4的同一性为87.6%，与小鼠PNAS - 4的同一性为85.5%。还总结了PNAS - 4蛋白的系统发育树。使用EGFP融合蛋白进行的细胞定位分析表明，xPNAS - 4定位于细胞质的核周区域。RT - PCR分析显示，xPNAS - 4作为母源表达基因，存在于早期胚胎发育的所有阶段。整体原位杂交表明，xPNAS - 4主要在外胚层和中胚层表达。此外，在体内显微注射xPNAS - 4 mRNA会导致发育缺陷，在非洲爪蟾胚胎中表现为小眼表型，在发育中的斑马鱼胚胎中表现为小眼或单眼表型。此外，显微注射xPNAS - 4反义吗啉代寡核苷酸（MOs）的胚胎表现出头部发育失败和轴缩短。
