The Effect Of Methylation In The CpG Island Of EYA2Gene On Occurrence And Development Of Lung Adenocarcinoma

The main purpose of this paper is to evaluate the methylation level of EYA2gene and its precise role in the tumorigenesis and development of lung adenocarcinoma. Firstly, western blotting and quantitative real-time PCR methods were used to detect the transcriptional and expression level of EYA2gene and results showed that the expression of EYA2protein was up-regulated in lung adenocarcinoma and A549lung cancer cells. Secondly, the data of bissulphited sequencing used to examine methylation level of entire CpG islands of EYA2gene suggested that the CpG islands of EYA2gene is abnormally hypomethylated in lung adenocarcinoma and there was a reversed trend between the expression level and the methylation level. Lastly, in order to figure out the impact of the expression level of EYA2gene on lung adenocarcinoma, EYA2gene was knocked down in lung adenocarcinoma cells with siRNA interference and some tumor development related-physiologic indices were also detected. Detailed protocol is as follows,1.Measuring cell proliferation rate by MTS method;2.After staining, examining cell cycle and apoptosis via flow cytometry;3.Observing cell migration and invasion condition through wound healing assay and trans-well approach respectively. The results exhibited that knockdown of EYA2gene suppressed the growth of tumour cells by attenuating cell proliferation, arresting the cell cycle in G0/G1phase, promoting cell apoptosis, and repressing cell migration and invasion. Therefore we concluded that, to some extent, abnormal hypomethylation and overexpression of EYA2gene lead to the occurrence and development of lung adenocarcinoma.