ABSTRACT
To investigate the role of Gli1 in DDP-resistance in lung cancer.Methods:A DDP-resistant human lung adenocarcinoma cell line A 549/DDP was cultured and transfected with Gli 1 siRNA.The mRNA and protein expression levels of Gli 1 were evaluated by qPT-PCR,Western blot analysis and immunofluorescence microscopy.The expression of Bcl-2、caspases-3、cyclinD1 was evaluated by Western blot analysis.Hoechst 33258 staining and flow cytometry were used to detect spontaneous cell apoptosis and cell cycle.The cell inhibition rate and DDP-induced cell death were examined by MTT and Annexin V-FITC/propidium iodide staining.Results:Gli1-knockdown by using Gli 1-specific siRNA led to a markedly decrease in Gli 1 mRNA and protein expression levels,when compared to negative siRNA transfected cells and untreated control cells (P=0.001).The downstream effectors of Gli1, Bcl-2 and cyclinD1 proteins were also inhibited (P=0.003),the expression of caspase-3 was increased (P=0.001).Hoechst 33258 staining showed that Gli1 depletion by Gli1 siRNA in A549/DDP cells could induce spontaneous apoptosis.The result of cell cycle showed Gli1 siRNA could lead more cells arrest in G1 phase.The IC50 of A549/DDP cells on DDP was (12.63 ±1.11 )μg/ml /(13.81±1.14)μg/ml,which decreased to (2.65±0.85)μg/ml after being transfected with Gli1 siRNA (P=0.000).Annexin V-FITC/propidium iodide staining showed that DDP-induced apoptosis rate in Gli 1-silencing cells was higher than that in negative siRNA or untreated control cells ( 35.19%±3.92% vs 6.43%±0.11%/5.01%±0.77%, P=0.000 ).Conclusion: Application of RNA interference can restrain the expression of Gli 1 mRNA and protein observably in A549/DDP cells,and increase the sensitivity of A549/DDP cells to cisplatin.Maybe Gli1 will become a new target to reverse the cisplatin resistance for lung cancer.
ABSTRACT
ObjectiveTo investigate the association between R405Q polymorphism of GLI1 gene and tetralogy of fallot(TOF).Methods In the case-control study,the R405Q polymorphism of GLI1 gene in 112 children with TOF and 200 healthy controls were detected with polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).The distribution of genotype and allele frequency at R405Q polymorphism site were analyzed and to investigate its relationship with the risk of TOF.ResultsThe distribution of genotype frequency at R405Q polymorphism site was not different between TOF group and the healthy control group(x2 =5.317 ,P = 0.07) .However, the distribution of allele frequency at R405Q polymorphism site was significantly different between TOF group and the healthy control group (x2 = 6.790, P = 0.009) , and the relative risk for TOF in A allele carriers was higher than that in G allele carriers (OR = 1.561,95％ CI 1.116 ～ 2.185) Conclusion The R405Q polymorphism of GLI1 gene is associated with TOF and people with A allele have higher risk with TOF.GLI1 gene might be the genetic susceptibility gene of TOF.
ABSTRACT
Objective In the candidate region 12q13 of simple congenital heart defect(CHD),two single nucleotide polymorphisms(SNPs)in the coding-region of GLI1 gene,rs11553626 and rs2228226,were chosen to investigate their distribution in 180 simple CHD patients and 200 normal controls which were collected in the first affliated hospital,China Medical University and The General Hospital of Shenyang Military Area,in order to determine the relationship between GLI1 gene and simple CHD.Methods Genotypes of these two SNPs were analyzed in 180 simple CHD patients and 200 normal controls by Denatured High Performance Liquid Chromatography(DHPLC)and sequencing from Jan.2000 to Jun.2005.?~2 test was applied to analyze the genotype frequency and allele frequency between CHD groups and control groups.Results No polymorphisms were found at rs11553626 while G/C polymorphisms were found at rs2228226.Remarkable significance was observed at rs2228226 in the allele frequency distribution(?~2=8.956,P