Effects And Molecular Mechanism Of Soy Foods Intake On Non-small Cell Lung Cancer

Objective: To study the relationship between intake of soy foods and the polymorphisms of EGFR and ESR2 genes with the incidence of lung cancer in non-smoking women.To explore the effect of daidzein and its three derivatives [A(4',7-diphenylsulfonyloxyisoflavone),B(4'-hydroxy-7-benzenesulfonyloxyisoflavone)and C(4'-acetoxy-7-benzenesulfonyloxyisoflavone)on proliferation of non-small cell lung cancer cells,and preliminary study on the mechanism of action of daidzein on non-small cell lung cancer cells.Methods: A case-control study was designed to investigate and use the Micro-ARRAY SNPs to detect the genetic polymorphisms of EGFR and ESR2 in case and control samples.EGFR and ESR2 gene polymorphisms and non-smoking women with lung cancer were analysis of the interaction between soy foods intake.Proliferation activity were detected by CCK-8 assay on lung cancer cells(A549 and H1299 cells)and normal lung cells(HELF cells)that is treat with daidzein and its derivatives in vitro.Apoptosis of cells were detected by Tunel or Flow Cytometry.The expression of m RNA was analyzed by human Array v4,4x180 K gene chip technology on H1299 cells which is treated with daidzein,and the enrichment of differentially expressed genes was analyzed by bioinformatics(GO,Pathway etc.).Then the expressions of m RNA and protein of apoptosis-related genes TP53 and Caspase9 in normal lung cells and lung cancer cells were verified by RT-q PCR and Western blot.Results: With the increase in soybean intake,the risk of lung cancer in women decreased by 0.85 [95% CI 0.70,1.28](P = 0.049).The risk of rs11534100 mutation in the EGFR gene and lung cancer in non-smoking women was 0.514 [95% CI 0.122,2.175](P = 0.006).Daidzein and its three derivatives can promote the proliferative activity of normal lung cells and inhibit the proliferative activity of lung cancer cells,and this effect has a tendency of concentration and time,what's more,daidzein can inhibit the apoptosis of normal lung cells and promote the apoptosis of lung cancer cells.Results of gene chip showed that there are 5 genes up-regulated and 35 genes down-regulated,and the results of significant enrichment of KEGG and GO indicated that the effects on H1299 was related to P53 pathway gene.After treatment with 10?M daidzein,results of RT-q PCR showed that the expression of TP53 in HELF,H1299 and A549 cells was 1.6 times [95% CI 1.12,2.14](P<0.05),2.3 times [ 95% CI 1.47,3.57](P < 0.05),1.5 times [95% CI 1.07,3.57](P < 0.05)higher than the control group respectively.Compared with the control group,the m RNA expression of Caspase9 in HELF and H1299 cells was 2.66 times [95% CI 1.56,2.03](P<0.05),2.7times [95% CI 1.56,4.97](P<0.05)respectively.The protein expression of Caspase9 in HELF and H1299 cells treated with 10 ?M daidzein was increased to 1.76 times[95% CI 1.58,1.93](P < 0.001)and 1.95 times [95% CI 1.89,2.01](P < 0.001)respectively,and the P53 protein was increased to 1.30 times [95% CI 1.14,1.47](P =0.005)and 1.49 times [95% CI 1.16,1.83](P = 0.036)respectively.After inhibiting the m RNA expression of TP53 by 10 ?M TP53 specific inhibitor Pifithrin-?,the m RNA and protein expression of TP53 in normal lung cells increased by 5.20 times[95% CI 1.05,9.34](P=0.032)and 1.77 times [95% CI 1.51,2.04](P=0.001),and no effects of daidzein on apoptosis of lung cancer cells was observed.Conclusion: soy foods intake can reduce the risk of lung cancer in non-smoking women.SNPs of EGFR are associated with the onset of lung cancer in non-smoking women.Daidzein and its three derivatives can inhibit the proliferation activity of lung cancer cells and increase the normal cell activity of lung,and promote the apoptosis of lung cancer cells and inhibit the apoptosis of normal lung cells.Daidzein can influence the proliferation of lung cancer cells by increasing the expression of m RNA and proteins of TP53 and Caspase9 in normal lung cells and lung cancer cells.The mechanism of the effect of daidzein on the proliferation and apoptosis of lung cancer cells and normal lung cells may be suggested.Related to the P53 pathway.