title: Methylation of neurofilament light polypeptide promoter is associated with cell invasion and metastasis in NSCLC.
abstract: The role of NEFL in NSCLC remains largely unknown. Immunohistochemistry was performed to investigate the expression of NEFL in 108 lung cancer specimens. NEFL expression was associated with decreased lymph node metastases and favorable prognosis. Furthermore, real-time PCR and Western blot were used to investigate the expression of the NEFL gene in NSCLC cell lines. Subsequently, lentivirus-mediated RNA interference and overexpression were used to demonstrate that knocked-down of NEFL enhanced the invasion and migration of A549 and H460 NSCLC cells, whereas NEFL overexpression resulted in a suppression of the invasion and migration of GLC-82 and L78 cells in vitro. In addition, bisulfite sequence PCR assay demonstrated that NEFL downregulation was associated with promoter methylation, and NEFL expression was restored after treatment with 5-Aza-dC. Finally, we demonstrated that NEFL inhibited the NF-κB pathway, thereby suppressing the expression of uPA and decreasing NSCLC invasiveness and migration. Our studies suggest that NEFL methylation is a novel mechanism for NSCLC invasion and metastasis and that NEFL may represent a candidate biomarker for recurrence and survival in patients with NSCLC.
authors.affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Department of Thoracic Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, PR China; Lung Cancer Research Center of Sun Yat-Sen University, Sun Yat-Sen University, Guangzhou 510275, PR China.
authors.forename: Zhuojian
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Department of Thoracic Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, PR China; Lung Cancer Research Center of Sun Yat-Sen University, Sun Yat-Sen University, Guangzhou 510275, PR China.
authors.forename: Baishen
authors.initials: B
authors.lastname: Chen
authors.affiliation: Department of Thoracic Surgery, General Hospital of Ningxia Medical University, Ningxia Medical University, Yinchuan 750004, PR China.
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: Gan
authors.affiliation: Department of Thoracic Surgery, Zengcheng People's Hospital, Sun Yat-Sen University, Zengcheng 511330, PR China.
authors.forename: Weicheng
authors.initials: W
authors.lastname: Hu
authors.affiliation: Department of Urology Surgery, Sun Yat-Sen Memorial Hospital, Guangzhou 510120, PR China.
authors.forename: Guangzheng
authors.initials: G
authors.lastname: Zhong
authors.affiliation: Department of Pathology, Sun Yat-Sen Memorial Hospital, Guangzhou 510120, PR China.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Li
authors.affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Department of Thoracic Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, PR China; Lung Cancer Research Center of Sun Yat-Sen University, Sun Yat-Sen University, Guangzhou 510275, PR China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Xie
authors.affiliation: Department of Pathology, Sun Yat-Sen Memorial Hospital, Guangzhou 510120, PR China.
authors.forename: Yeqing
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pathology, Sun Yat-Sen Memorial Hospital, Guangzhou 510120, PR China.
authors.forename: Haigang
authors.initials: H
authors.lastname: Li
authors.affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Department of Thoracic Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, PR China; Lung Cancer Research Center of Sun Yat-Sen University, Sun Yat-Sen University, Guangzhou 510275, PR China.
authors.forename: Xia
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Oral and Maxillofacial Surgery, Sun Yat-Sen Memorial Hospital, Guangzhou 510120, PR China. Electronic address: zhiquanhuang1978@126.com.
authors.forename: Zhiquan
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Department of Thoracic Surgery, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, PR China; Lung Cancer Research Center of Sun Yat-Sen University, Sun Yat-Sen University, Guangzhou 510275, PR China. Electronic address: dydchenju@163.com.
authors.forename: Ju
authors.initials: J
authors.lastname: Chen
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000473:pathology / Q000556:secondary*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology*; D009361:Neoplasm Invasiveness; D016900:Neurofilament Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*
chemical_list: D016900:Neurofilament Proteins; C044882:neurofilament protein L
keywords: NEFL; NF-κB; NSCLC; methylation; uPA