Methylation of CLEC14A is associated with its expression and lung adenocarcinoma progression

Abstract

Our main objective is probing the effect of methylation of CLEC14A on its expression and lung adenocarcinoma (LUAD) progression. Microarray analysis was utilized to screen out differentially downregulated genes with hypermethylation in LUAD tissues. The CLEC14A expression level was measured by western blot analysis and qRT-PCR. Methylation-specific-PCR was performed to evaluate methylation status of CLEC14A. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromid (MTT) assay was used to check the relation between CLEC14A expression and cell proliferation. Cell cycle, cell apoptosis, migration, and invasion were respectively detected by the flow cytometry assay, wound healing assay, and transwell assay. Tumor xenograft models were established for investigating the effect of CLEC14A on tumor formation. CLEC14A expression in LUAD tissues was impaired compared with that in adjacent tissues, and CLEC14A promoter was highly methylated in LUAD. Overexpressing CLEC14A or inhibiting the methylation level of CLEC14A in A549 and LTEP-a-2 cells impeded the duplication of LUAD cells, promoted apoptosis, attenuated cell migration, and invasion ability, and arrested cell cycle at the G0/G1 phase. Overexpression of CLEC14A inhibited tumorigenesis of LUAD cells in nude mice. The promoter of CLEC14A is methylated in LUAD, leading to downregulation of CLEC14A in LUAD. CLEC14A acts as an antitumor role in LUAD by suppressing cell proliferation, migration, invasion, promoting cell apoptosis, and reducing tumorigenicity in nude mice. Thus, the inhibition of CLEC14A methylation is a novel strategy for the clinic treatment of LUAD.

Serum ROBO4 and CLEC14A: preliminary evaluation as diagnostic and progression biomarkers in colorectal cancer patients

“…Simmilary, in non-small lung cancer, high ROBO4 tissue expression was related to good prognosis and was connected with normalization of endothelial cells and reduction of cancer spread [16]. Considering CLEC14A, recent reports indicate that elevated levels of this molecule can inhibit carcinogenesis and progression of lung adenocarcinoma [35]. The expression of ROBO4 or CLEC14A molecules in various cancers tissues (up-or down-regulation) suggests that these proteins may act as important modulators of tumorgenesis and tumor progression.…”

Serum ROBO4 and CLEC14A: preliminary evaluation as diagnostic and progression biomarkers in colorectal cancer patients

“…Simmilary, in nonsmall lung cancer, high ROBO4 tissue expression was related to good prognosis and was connected with normalization of endothelial cells and reduction of cancer spread [16]. Considering CLEC14A, recent reports indicate that elevated levels of this molecule can inhibit carcinogenesis and progression of lung adenocarcinoma [35]. The expression of ROBO4 or CLEC14A molecules in various cancers tissues (up-or down-regulation) suggests that these proteins may act as important modulators of tumorgenesis and tumor progression.…”

C‐type lectin domain group 14 proteins in vascular biology, cancer and inflammation

“…Interestingly, high expression of CLEC14A in this cancer type correlated with improved clinical outcomes. A further study indicated that the methylation status of CLEC14A strongly correlated with its expression levels in NSCLC, and CLEC14A protein levels were reduced in tumour tissues compared to healthy adjacent tissue . Similar to CD93, CLEC14A was described as a key gene in a proposed ‘tumour angiogenesis signature’ determined by meta‐analysis of over 1000 tumour samples including breast, renal and head and neck cancers .…”

“…CLEC14A expression has been demonstrated in two different human lung cancer cell lines in vitro and when CLEC14A was further overexpressed in these cell lines, this led to reductions in proliferation, migration and invasion as well as reductions of in vivo tumour formation as xenografts in nude mice . However, the physiological relevance of CLEC14A expression in tumour cells themselves remains to be seen as such expression has not been reported in clinical specimens or in tumour cells by any other group.…”