Detection Of Methylation Of SLC35F2 Promoter Region In Non-small Cell Lung Cancer And Screening Of Related Differentially Expressed Genes

Lung cancer is the malignant tumor with the highest morbidity and mortality.Studies have found that 35 member F2 of the solute carrier family(SLC35F2)is highly expressed in lung cancer tissues,But the expression mechanism causing high expression of this gene has not been reported.DNA methylation can be involved in the regulation of the expression of various genes,So in this experiment,by collecting cancer tissues and adjacent tissues from patients with non-small cell lung cancer,q PCR selected 10 pairs of cancer tissues and adjacent tissues with relatively high expression of SLC35F2,Using bisulfite amplicon sequencing technology to detect the methylation of gene promoter region in SLC35F2 highly expressed tissue,To evaluate whether the degree of methylation of the SLC35F2 promoter region affects its expression regulation in lung cancer and whether it can be used as a biomarker for lung cancer diagnosis or prognosis.A preliminary study of the cell function found that,in the non-small cell lung cancer cell line with low expression of SLC35F2,the proliferation,migration and invasion ability of the cells were significantly reduced,and the level of apoptosis increased,which was in line with the regulation of proto-oncogenes.Therefore,this experiment will use gene chip technology to screen out the differentially expressed genes of non-small cell lung cancer cell lines with low expression of SLC35F2 and control cell lines.And selected genes related to proliferation for q PCR and WB detection to verify the reliability of chip data,and provide a basis for further analysis of the biological processes that the gene may be involved.Methylation results showed that the average methylation ratio and methylation ratio of each site in the tumor group and normal group were less than 0.1,and the degree of methylation was not significant.Compared between groups,the average methylation degree of the tumor group was significantly lower than that of the normal group,and the methylation degree of the tumor group at positions 67,70,and 190 was significantly lower than that of the normal group.The results of gene chip showed that the A549 cell line with low expression of SLC35F2 constructed by lentiviral vector had 326 up-regulated genes and 659 down-regulated genes relative to the control group.QPCR and WB detection of CHEK1,CDK6,CDKN1 A,PLK1,SMAD2,Theresults showed that the changes in m RNA levels and protein levels were consistent with the chip results,proving the reliability of the chip data.The analysis of the results concluded that the methylation of the SLC35F2 promoter region is not suitable as a new target for the treatment of non-small cell lung cancer.The next step will continue to study the regulatory influence of epigenetics,aiming to find a new target for the treatment of non-small cell lung cancer.From the differential genes screened in SLC35F2 low-expressing non-small cell lung cancer cell lines,we will first select the genes most likely to affect the occurrence and development of non-small cell lung cancer,and continue to further study their expression levels and related signals path in non-small cell lung cancer.Secondly,using bioinformatics to analyze the expression regulation of differentially expressed genes in different tumors,and further assess whether it is related to non-small cell lung cancer.