Effect of TRAF6 Downregulation on Malignant Biological Behavior of Lung Cancer Cell Lines
10.3779/j.issn.1009-3419.2015.11.01
- VernacularTitle:下调TRAF6表达对肺癌细胞株恶性生物学行为的影响
- Author:
LIN GEN
1
;
HUANG CHUANGZHONG
;
SU GUANGJIAN
;
HU HUIHUA
;
XU HAIPENG
;
HUANG CHENG
Author Information
1. 350014 福州，福建医科大学教学医院，福建省肿瘤医院胸部肿瘤内科
- Keywords:
Lung neoplasms;
Tumor necrosis factor receptor-associated factor 6;
Nuclear factor-kappa B;
Apopto-sis;
Invasion
- From:
Chinese Journal of Lung Cancer
2015;(11):661-667
- CountryChina
- Language:Chinese
-
Abstract:
Background and objectiveIt has been proven that tumor necrosis factor receptor-associated factor 6 (TARF6) was a commonly ampliifed oncogene in lung cancer. However, the precise role of TARF6 protein in lung cancer has not been extensively investigated. hTis study analyzed the effects of TARF6 on the proliferation, apoptosis, cell cycle, migration, and invasion capability of lung cancer cell lines, as well as the potential molecular mechanisms involved.MethodsTo address the expression of TARF6 in lung cancer cells, four lung cancer cell lines (A549, H1650, SPC-A-1 and Calu-3) were assayed to determine the expression of TARF6 protein by Western blot and TARF6 mRNA via qRT-PCR. Moreover, siRNA targeting TARF6 was introduced into SPC-A-1 and Calu-3 cells. Nuclear factor-?B (NF-?B) DNA-binding activity, apoptosis rates, cell proliferation, cell cycle, migration, and invasion were determined by electrophoretic mobility shitf assay, lfow cytometry, MTS assay, lfow cytometry, scratch test, and transwell chamber assay, respectively. Western blot analysis was also performed to evalu-ate the expression of the following proteins through K63-ubiquitination: P65, CD24 and CXCR4. Whole-genome sequencing analysis was conducted using a second-generation sequencer in SPC-A-1 cells.Results TARF6 was highly up-expressed in SPC-A-1 and Calu-3 cell lines than the other two cells, which also showed K63-ubiquitinization in TARF6. However, consti-tutive activation of NF-?B was observed only in SPC-A-1 lung cancer cells. Downregulation of TARF6 suppressed the NF-κB activation, cell migration, and invasion but promoted the cell apoptosis of SPC-A-1 cells. Markedly decreased expression of CD24 and CXCR4 was observed in SPC-A-1 cells transfected by TARF6 siRNA. Nevertheless, TARF6 downregulation did not affect the proliferation and cell cycle of SPC-A-1 cells. Additionally, TARF6 regulation did not affect the proliferation, apoptosis, cell cycle, migration, and invasion of Calu-3 cells. No mutations and no changes in gene copy numbers of TARF6 were found by whole-exome sequencing of SPC-A-1 cells.ConclusionTARF6 may be involved in cell migration, invasion, and apoptosis of SPC-A-1 cells, possibly through regulating the NF-?B-CD24/CXCR4 pathway.
