Park Gun-Hoo, Lee Su-Jin, Yim Hyunee, Han Jae-Ho, Kim Hyon J, Sohn Young-Bae, Ko Jung Min, Jeong Seon-Yong
Department of Medical Genetics, Ajou University School of Medicine, Suwon, Republic of Korea.
Department of Pathology, Ajou University School of Medicine, Suwon, Republic of Korea.
Oncol Rep. 2014 Oct;32(4):1347-54. doi: 10.3892/or.2014.3379. Epub 2014 Aug 4.
Neurofibromatosis type 1 (NF1) caused by NF1 gene mutation is a commonly inherited autosomal dominant disorder. Malignant peripheral nerve sheath tumors (MPNSTs), a type of aggressive sarcoma, are a major cause of mortality in NF1 patients. The malignant transformation of benign plexiform neurofibromas (PNs) to MPNSTs is a marked peculiarity in NF1 patients, yet the pathogenesis remains poorly understood. We found that an actin-associated protein transgelin (SM22) was highly expressed in NF1-deficient MPNST tissues compared to NF1-deficient PN tissues using immunohistological staining and primary cultured MPNST cells in western blot analysis. We further found that this transgelin upregulation was caused by increased transcriptional expression of the TAGLN gene encoding transgelin. Comparison of DNA methylation values in the promoter and subpromoter regions of the TAGLN gene in three types of NF1-deficient primary-cultured cells, derived from an NF1 patient's normal phenotype, a benign PN and MPNST tissues, revealed that the TAGLN gene was hypomethylated in the MPNST cells. Next, to determine the functional role of transgelin in MPNST pathogenesis, we manipulated the TAGLN gene expression and investigated the alteration of the RAS-mitogen-activated protein kinase (MAPK) signaling pathway in the normal-phenotypic and malignant tumor cells. The downregulation of TAGLN expression in NF1-deficient MPNST tumor cells through the treatment of the small interfering RNA resulted in a decrease in the RAS activation (GTP-RAS) and the downstream ERK1/2 activation (phosphorylated ERK1/2), while the overexpression of TAGLN in normal-phenotypic NF1-deficient cells caused an increase in RAS and ERK1/2 activation. These results indicate that upregulation of transgelin caused by hypomethylation of the TAGLN gene is closely involved in tumor progression in NF1.
由NF1基因突变引起的1型神经纤维瘤病(NF1)是一种常见的常染色体显性遗传病。恶性外周神经鞘瘤(MPNST)是一种侵袭性肉瘤,是NF1患者死亡的主要原因。良性丛状神经纤维瘤(PN)向MPNST的恶性转化是NF1患者的一个显著特征,但其发病机制仍知之甚少。通过免疫组织化学染色以及蛋白质免疫印迹分析原代培养的MPNST细胞,我们发现与NF1缺陷的PN组织相比,肌动蛋白相关蛋白转胶蛋白(SM22)在NF1缺陷的MPNST组织中高表达。我们进一步发现,这种转胶蛋白的上调是由编码转胶蛋白的TAGLN基因转录表达增加所致。比较来自NF1患者正常表型、良性PN和MPNST组织的三种NF1缺陷原代培养细胞中TAGLN基因启动子和亚启动子区域的DNA甲基化值,发现TAGLN基因在MPNST细胞中低甲基化。接下来,为了确定转胶蛋白在MPNST发病机制中的功能作用,我们操纵TAGLN基因表达,并研究正常表型和恶性肿瘤细胞中RAS-丝裂原活化蛋白激酶(MAPK)信号通路的改变。通过小干扰RNA处理使NF1缺陷的MPNST肿瘤细胞中TAGLN表达下调,导致RAS激活(GTP-RAS)和下游ERK1/2激活(磷酸化ERK1/2)减少,而在正常表型的NF1缺陷细胞中过表达TAGLN则导致RAS和ERK1/2激活增加。这些结果表明,TAGLN基因低甲基化导致的转胶蛋白上调与NF1中的肿瘤进展密切相关。