Department of Neurosurgery, Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China.
Brain Res. 2013 Jul 12;1521:68-78. doi: 10.1016/j.brainres.2013.05.021. Epub 2013 May 20.
Paired box 3 (PAX3) is overexpressed in glioma tissues compared to normal brain tissues, however, the pathogenic role of PAX3 in human glioma cells remains to be elucidated. In this study, we selected the human glioma cell lines U251, U87, SHG-44, and the normal human astrocytes, 1800, which have differential PAX3 expression depending upon the person. SiRNA targeting PAX3 and PAX3 overexpression vectors were transfected into U87 and SHG-44 glioma cell lines, and cell proliferation, invasion, apoptosis, and differentiation were examined by CCK-8 assays, transwell chamber assays, tunnel staining, Annexin V/PI analysis, and Western blotting, respectively. In addition, we used subcutaneous tumor models to study the effect of PAX3 on the growth of glioma cells in vivo. We found that PAX3 was upregulated in the three glioma cell lines. PAX3 knockdown inhibited cell proliferation and invasion, and induced apoptosis in the U87MG glioblastoma cell line, whereas PAX3 upregulation promoted proliferation, inhibited apoptosis, and increased invasion in the SHG-44 glioma cell line. Moreover, we found that targeting PAX3 expression in glioma cell lines together with chemotherapeutic treatment could increase glioma cell susceptibility to the drug. In subcutaneous tumor models in nude mice using glioma cell lines U-87MG and SHG-44, inhibition of PAX3 expression in glioblastoma U-87MG cells suppressed tumorigenicity, and upregulation of PAX3 expression in glioma SHG-44 cells promoted tumor formation in vivo. These results indicate that PAX3 in glioma is essential for gliomagenesis; thus, targeting PAX3 or its downstream targets may lead to novel therapies for this disease.
配对盒基因 3（PAX3）在胶质瘤组织中过度表达，而在正常脑组织中则不表达，然而，PAX3 在人类神经胶质瘤细胞中的致病作用仍有待阐明。在这项研究中，我们选择了人类神经胶质瘤细胞系 U251、U87、SHG-44 和正常人类星形胶质细胞 1800，这些细胞系根据个体的不同而具有不同的 PAX3 表达水平。靶向 PAX3 的 siRNA 和 PAX3 过表达载体被转染到 U87 和 SHG-44 神经胶质瘤细胞系中，通过 CCK-8 检测、Transwell 小室检测、隧道染色、Annexin V/PI 分析和 Western blot 分别检测细胞增殖、侵袭、凋亡和分化。此外，我们还使用皮下肿瘤模型来研究 PAX3 对体内神经胶质瘤细胞生长的影响。我们发现 PAX3 在三种神经胶质瘤细胞系中上调。PAX3 敲低抑制 U87MG 神经母细胞瘤细胞系的细胞增殖和侵袭，并诱导细胞凋亡，而 PAX3 上调促进 SHG-44 神经胶质瘤细胞系的增殖、抑制凋亡和增加侵袭。此外，我们发现靶向神经胶质瘤细胞系中的 PAX3 表达与化疗药物治疗相结合可以增加神经胶质瘤细胞对药物的敏感性。在使用神经胶质瘤细胞系 U-87MG 和 SHG-44 的裸鼠皮下肿瘤模型中，抑制神经母细胞瘤 U-87MG 细胞中的 PAX3 表达抑制了肿瘤发生，上调神经胶质瘤 SHG-44 细胞中的 PAX3 表达促进了体内肿瘤的形成。这些结果表明，PAX3 在神经胶质瘤中对于神经胶质瘤的发生是必不可少的；因此，靶向 PAX3 或其下游靶标可能为该疾病提供新的治疗方法。
