Cancer Management and Research (Dec 2018)
Chronic cadmium exposure aggravates malignant phenotypes of nasopharyngeal carcinoma by activating the Wnt/β-catenin signaling pathway via hypermethylation of the casein kinase 1α promoter
Abstract
Lin Peng,1,2 Yi-Teng Huang,3 Fan Zhang,4 Jiong-Yu Chen,4 Xia Huo5 1Clinical Laboratory, Cancer Hospital of Shantou University Medical College, Shantou 515041, People’s Republic of China; 2Laboratory of Environmental Medicine and Developmental Toxicology, Shantou University Medical College, Shantou 515041, People’s Republic of China; 3Health Care Center, The First Affiliated Hospital of Shantou University Medical College, Shantou 515041, People’s Republic of China; 4Oncological Research Lab, Cancer Hospital of Shantou University Medical College, Shantou 515031, People’s Republic of China; 5Laboratory of Environmental Medicine and Developmental Toxicology, Guangzhou and Guangdong Key Laboratory of Environmental Pollution and Health, School of Environment, Jinan University, Guangzhou 510632, People’s Republic of China Background: Our previous study has shown that cadmium (Cd) exposure is not only a risk factor for nasopharyngeal carcinoma (NPC), but also correlated with the clinical stage and lymph node metastasis. However, the underlying molecular events of Cd involved in NPC progression remain to be elucidated. Purpose: The objective of this study was to decipher how Cd impacts the malignant phenotypes of NPC cells. Methods: NPC cell lines CNE-1 and CNE-2 were continuously exposed with 1 μM Cd chloride for 10 weeks, designating as chronic Cd treated NPC cells (CCT-NPC). MTT assay, colony formation assay and xenograft tumor growth were used to assess cell viability in vitro and in vivo. Transwell assays were performed to detect cell invasion and migration. The protein levels of E-cadherin, N-cadherin, Vimentin as well as β-catenin and casein kinase 1α(CK1α) were measured by Western blot. Immunofluorescence staining was used to observe the distribution of filament actin (F-actin), β-catenin and CK1α. The mRNA levels of downstream target genes of β-catenin were detected by RT-PCR. Wnt/β-catenin signaling activity was assessed by TOPFlash/FOPFlash dual luciferase report system. MS-PCR was used to detect the methylation status of CK1α. Finally, the activation of Wnt/β-catenin pathway and cell biological properties were examined following treatment of CCT-NPC cells with 5-aza-2-deoxy-cytidine(5-aza-CdR). Results: CCT-NPC cells showed an increase in cell proliferation, colony formation, invasion and migration compared to the parental cells. Cd also induced cytoskeleton reorganization and epithelial-to-mesenchymal transition. Upregulation and nuclear translocation of β-catenin and increased luciferase activity accompanied with transcription of downstream target genes were found in CCT-NPC cells. Treatment of CCT-CNE1 cells with 5-aza-CdR could reverse the hypermethylation of CK1α and attenuate the cell malignancy. Conclusion: These results support a role for chronic Cd exposure as a driving force for the malignant progression of NPC via epigenetic activation of the Wnt/β-catenin pathway. Keywords: cadmium, nasopharyngeal carcinoma, Wnt/β-catenin, DNA methylation, casein kinase 1α
