Hypomethylation-induced expression of S100A4 increases the invasiveness of laryngeal squamous cell carcinoma
Hypomethylation-induced expression of S100A4 increases the invasiveness of laryngeal squamous cell carcinoma
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DOI：
10.3892/or_00000738
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发表时间：
2010-04-01
期刊：
影响因子：
4.2
通讯作者：
Fu, Wei-Neng
中科院分区：
文献类型：
--
作者：
Liu, Jia;Guo, Yan;Fu, Wei-Neng
本研究的目的是鉴定5AZA CdR在喉鳞状细胞癌（LSCC）中的相关基因，并探讨S100 A4在喉鳞状细胞癌发生、发展中的作用。应用双向电泳结合MALDI-TOF-MS技术鉴定5 AZA CdR诱导的Hep-2细胞中差异表达蛋白，RT-PCR、Western blotting和甲基化特异性PCR分别检测S100 A4基因mRNA、蛋白水平和DNA甲基化状态。transwell迁移实验检测siRNA S100 A4转染的Hep-2细胞的侵袭能力。结果显示，5 AZA-CdR处理后，Hep-2细胞的蛋白质表达谱有明显的差异，其中包括S100钙结合蛋白A4（S100 A4）。RT-PCR和Western blotting结果显示，S100 A4在转移淋巴结中的mRNA和蛋白水平均显著高于癌旁正常喉组织和癌旁正常喉组织。DNA甲基化状态在喉鳞状细胞癌和肺腺癌组织中有显著差异。RNA干扰S100 A4后Hep-2细胞中S100 A4的表达水平下降。与对照组中的细胞数相比，RNAi S100 A4组中穿过基底膜滤器的细胞数显著较低。在所有DNA甲基转移酶抑制剂处理的Hep-2细胞中鉴定的S100 A4的异常表达似乎是由S100 A4的异常DNA甲基化状态引起的。S100 A4的异常表达改变了喉鳞癌的侵袭性。
The present study aimed to identify genes related to 5AZA-CdR in laryngeal squamous cell carcinoma (LSCC) and to investigate the role of S100A4 in the development and aggression of LSCC. Differentially expressed proteins were identified in Hep-2 cells treated With 5AZA-CdR by two-dimensional gel electrophoresis combined With MALDI-TOF-MS. mRNA, protein levels and DNA methylation status Of S100A4 were assessed by RT-PCR, Western blotting and methylation-specific PCR, respectively. The invasiveness of Hep-2 cells transfected by siRNA S100A4 was determined by transwell migration assay. Protein profiles from Hep-2 cells treated with 5AZA-CdR were obtained, and several differentially expressed proteins Such as S100 calcium-binding protein A4 (S100A4) were identified. Results of RT-PCR and Western blotting revealed that both mRNA and protein levels of S100A4 were significantly higher ill the metastatic lymph nodes than those in paired adjacent normal laryngeal (PANL) or tumor tissues. The DNA methylation status displayed significant differences between the LSCC and the PANL tissues. The expression level of S100A4 decreased in Hep-2 Cells undergoing RNA interference of S100A4. The number of cells which crossed the basement membrane filter was significantly lower in the RNAi S100A4 group when compared with file number in the control group. The abnormal expression of S100A4 identified in Hep-2 cells treated with all inhibitor of DNA methyltransferase appeared to result from the aberrant DNA methylation status of S100A4. The abnormal expression of S100A4 altered the invasiveness of LSCC.