Effect of ITGA6 gene on non-small cell lung cancer brain metastasis cells
Sun Xiaoyang
Gu Qiang
Yao Jiahao
Chen Dongxia
Liang Peng
Abstract：Objective To deeply investigate the specific mechanisms of ITGA6 gene in the processes of proliferation,cell cycle regulation,invasion,and migration of non-small cell lung cancer(NSCLC)brain metastasis cells.Methods By mining the NSCLC brain metastasis-related data from the GEO database,this study successfully identified ITGA6 as an important factor associated with the occurrence and progression of NSCLC brain metastasis.Based on this,the correlation between the expression level of ITGA6 gene and the prognosis of NSCLC brain metastasis patients was further explored.To verify the expression pattern of ITGA6 in NSCLC brain metastasis,real-time quantitative reverse transcription PCR(qRT-PCR)and Western blot techniques were employed to measure the expression level of ITGA6 in NSCLC brain metastasis cell lines.Using gene transfection technology,negative control(NC group)and ITGA6-specific silencing vector(si-ITGA6 group)were transfected into the NSCLC brain metastasis cell lines H1915-BrM and A549-BrM in vitro.The proliferative ability of the cells was assessed by CCK-8 proliferation assay.The EDU assay was used to further verify the proliferative changes.The flow cytometry was applied to detect the distribution of cell cycle.Transwell invasion assay and wound healing assay were used to analyze the invasion and migration abilities of the cells.Western blotting was used to detect the expression levels of epithelial-mesenchymal transition(EMT)-related proteins.Results Differentially expressed genes in NSCLC brain metastasis were systematically analyzed using the GEO database,revealing ITGA6 as a key upregulated gene.Clinical data analysis indicated that high expression of ITGA6 was significantly associated with poor prognosis in NSCLC brain metastasis patients(P<0.05).Further verification by qRT-PCR and Western blot techniques confirmed the high expression of ITGA6 in lung cancer brain metastasis cell lines.The results of CCK-8 and EDU assays showed that si-ITGA6 significantly inhibited the proliferation of NSCLC brain metastasis cells H1915-BrM and A549-BrM(P<0.05).Flow cytometry revealed that after si-ITGA6 treatment,the proportion of cells in the G0/G1 phase increased,while cells in the S and G2 phases decreased,indicating cell cycle arrest(P<0.05).Both Transwell and wound healing assays showed that si-ITGA6 significantly reduced the invasion and migration abilities of NSCLC brain metastasis cells.Western blot analysis revealed that the expression of the epithelial marker E-cadherin was significantly increased in the si-ITGA6 group,while the expression of mesenchymal markers Vimentin and FN,and matrix metalloproteinases(MMP-2 and MMP-9),was significantly decreased,suggesting inhibition of the EMT process.Conclusions Specific silencing of the ITGA6 gene can significantly inhibit the proliferation,migration,and invasion abilities of NSCLC brain metastasis cells,arrest the cell cycle,and regulate the expression of EMT-related proteins.
Keywords：brain neoplasmsneoplasm metastasislung cancernon-small cellintegrin α6
Publication Date：2024-08-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：9( 485-493 )