ABSTRACT
OBJECTIVE@#To investigate the association between-1304T/G polymorphism in the promoter of MKK4 gene and the susceptibility in sporadic nasopharyngeal carcinoma.@*METHOD@#MKK4-1304T/G genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 90 NPC cases and 30 healthy controls.@*RESULT@#The number of nasopharyngeal carcinoma patients carrying with TG+GG genotype was much higher than those of controls (82.2% vs 66.7%, χ² =10.076, P < 0.05). Analysis showed that compared with the-1304TT genotype, -1304TG heterozygous reduced risk of nasopharyngeal carcinoma 0.56 fold (95% CI = 0.164-1.178, P < 0.01) and-1304GG lower 0.58 fold (95% CI = 0.126-1.381, P < 0.01), TG+ GG genotype variation risk of nasopharyngeal carcinoma decreased 0.72 fold (95% CI = 0.105-0.753, P < 0.01).@*CONCLUSION@#MKK4 gene-1304TG genotype can reduce risk of nasopharyngeal carcinoma, and it may be an independent protection factor in sporadic nasopharyngeal carcinoma.
Subject(s)
Humans , Carcinoma , Genetic Predisposition to Disease , Genotype , Heterozygote , MAP Kinase Kinase 4 , Genetics , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms , Genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Promoter Regions, GeneticABSTRACT
Objective:Discussion MKK4 protein expression in nasopharyngeal carcinoma and -1044 A/T polymorphism correlation.Methods:90 patients with nasopharyngeal carcinoma , MKK4 protein expression was determined by immunohistochemical staining,polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP ) to detect the gene -1044A/T sites monocytes nucleotide polymorphism.Results:MKK4 protein expression in nasopharyngeal carcinoma (-) was 24.4%(22/90),(+) was 15.6%(14/90),(++) was 34.4%(31/90),(+++) was 25.6% (23/90).Low expression (-/+) patients with a total of 36 cases,-1044AA genotype accounted for 22 cases (61.11%),AT genotype accounted for 12 cases (33.33%),TT genotype accounted for two cases (5.56%),AT+TT gene type accounted for 14 cases (38.89%).The patients with high MKK4 expression of 54 cases,of which accounted for 38 cases of AA genotype (70.37%),AT genotype accounted with 15 cases (27.78%),TT genotype accounted for one case (1.85%),AT +TT genotype accounted for 16 cases (29.63%).Low expression and high expression of T gene mutation occurs no significant ( Z=0.323 , P=0.747 ) .Conclusion: MKK4 protein expression correlated with -1044 A/T gene promoter polymorphisms was no significant correlation .
ABSTRACT
OBJECTIVE@#To investigate the effect of gene silencing of Bmi-1 on proliferation regulation of CD44+ nasopharyngeal carcinoma cancer stem-like cells (CSC-LCs).@*METHOD@#The sequence-specific short hairpin RNA lentivirus targeting at human Bmi-1 gene (LV-Bmi-1shRNA) was constructed and was used to infect CD44+ nasopharyngeal carcinoma cells which were sorted by flow cytometry. A lentiviral which included a random sequence was also designed to serve as a negative control. We employed fluorescence microscope and flow cytometry to detect infection efficiency; real-time PCR was used to detect Bmi-1 and its downstream gene while each protein expression level was confirmed by western blotting protocol; CCK-8 proliferation assay was applied to measure proliferation capacity; tumor spheroid assay was used to evaluate the self-renewal capacity. Colony formation assay was used to measure cell colony formation capability; flow cytometry analyzed cell cycle distribution.@*RESULT@#The constructed LV-Bmi-1shRNA successfully infected into the CD44+ nasopharyngeal carcinoma cells. The infection efficiency could reach above 95%; LV-Bmi-lshRNA effectively inhibited Bmi-1 mRNA and protein expression, while the downstream gene p16INK4a and p14ARF mRNA as well as protein expression level were upregulated (P < 0.05). Notablely, the proliferation, colony formation, self-renewal capabilities of the experimental group decreased significantly (P < 0.05). In addition, the cell cycle arrested at the G0-G1 phase.@*CONCLUSION@#Gene silencing of Bmi-1 inhibited the proliferation, colony formation and self-renewal capabilities of the CD44+ nasopharyngeal carcinoma CSC-LCs, inhibited the cell cycle processes, which may mediate through Bmi1-p16INK4a/p14ARF-p53 pathway. Our experimental results indicated that Bmi-1 gene may play an important role in the maintenance of the stem cell-like characteristics of CD44+ nasopharyngeal carcinoma cells. Bmi-1 gene may be a potential new target for the treatment of nasopharyng al carcinoma in the future.
Subject(s)
Humans , Carcinoma , Cell Cycle , Cell Division , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p16 , Metabolism , Gene Silencing , Hyaluronan Receptors , Metabolism , Lentivirus , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms , Genetics , Pathology , Neoplastic Stem Cells , Cell Biology , Polycomb Repressive Complex 1 , Genetics , RNA, Messenger , RNA, Small Interfering , Tumor Suppressor Protein p14ARF , Metabolism , Tumor Suppressor Protein p53 , MetabolismABSTRACT
Objective:To investigate the expression of MKK 4 protein in the nasopharyngeal carcinoma and its clinical significance.Methods:Immunohistochemical methods were employed to analyze MKK 4 positive expression intensity and positive cells in freshly collected nasopharyngeal carcinoma of both 90nasopharyngeal carcinoma cases and 20 chronic nasopharyngitis control.The clinical pathological characteristic were analyzed.Results:The data obtained by MKK4 immunohistochemistry showed that the MKK 4 positive rate was higher in control group than in the NPC group (95.5%vs 75.6%,P0.05 ) . Conclusion:Positive rate of MKK4 protein in nasopharyngeal carcinoma tissues is lower than in chronic nasopharyngitis.MKK4 protein expressions in nasopharyngeal carcinoma tissues negatively correlated with lymph node metastasis ,clincal stage ,invasive depth ,and TTP (Time to progression),but not with age,gender,location and tumor volume.