Expreddion Of Snail In Lung Adenocarcinoma Stem Cells And Its Correlation With Cisplatin Resistance

Objective: To explore the ability of secondary ball formation and differentiation of lung adenocarcinoma cells.The expression of Snail protein in lung cancer cell lines and cisplatin-resistant lung cancer cell lines was studied.Methods: 1.The expression of SNAI1 gene in non-small cell lung cancer cell lines was analyzed by GEO database.2.The tumor cells of lung adenocarcinoma A549 cell line were cultured by serum-free suspension culture technique.The second generation tumor cell pellet was confirmed by the second generation of spheroids,induced differentiation and immunofluorescence detection of stem cell markers CD133 and Sca-1 has stem cell characteristics and can be used as a tumor stem cell model.3.The expression of Snail protein in lung cells of lung adenocarcinoma cell line and the expression of Snail protein and stem cell markers CD133 and Sca-1 in A549 / DDP cell line were detected by immunofluorescence staining protein expression.Results: 1.The expression of SNAI1 gene in the lung adenocarcinoma A549 cell line was detected in the GDS4296 database.The expression of SNAI1 in cisplatin-resistant non-small cell lung cancercell line H460 / DDP was detected in GSE21656 data.Expression was higher than H460 cell line,statistically significant(P <0.05).The expression of SNAI1 gene in lung cancer stem cells isolated from H460 cell line in GSE54712 database was higher than that in differentiated cells.2.The use of serum-free suspension culture technology,can be cultured primary tumor cell ball.And the second generation of tumor cell spheres can be obtained by dissociating the cultured primary tumor cell globules into individual cells and continuing with the incubation of the stem cells(SCM),indicating that the primary tumor cells have the ability to double the ball.The cells of the second generation tumor cells were induced to differentiate.After incubation for 36 hours,cells adhered to the bottom of the culture flask were observed,indicating that the second generation of cultured tumor cells had differentiation ability.The expression of CD133 and Sca-1 in the stem cells of tumor stem cells were detected by immunofluorescence staining in the cytoplasm of tumor cells.3.The results of immunofluorescence staining showed that the expression of Snail protein in the second generation tumor cells was positive in the nucleus.The positive expression rate of Snail in A549 cell line was 5%,the positive expression rate in A549 / DDP was 15%Statistically significant(P <0.05).The positive expression rate of CD133 in A549 cell line was 5.6%,and the positive expression rate of A549 /DDP was 14.7%,which was statistically significant(P <0.05).The positive expression rate of Sca-1 in A549 cell line was 12.6%,and the positive expression rate of A549 / DDP was 15%,which was not statistically significant(P> 0.05).Conclusion : Snail protein is highly expressed in the second generation of lung adenocarcinoma stem cells enriched by A549 cell lines and is associated with cisplatin resistance.