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Objective To study the effects of deoxycytidine (5-aza-2 deoxycytidine, DAC) on DNA Methylation state and expression of mRNA and protein of pl6 gene in human squamous lingual carcinoma SCC-9 cells in vitro. Methods The SCC-9 cells were divided into four groups, group 0, 1, 2 and 3 which processed using three gradients concentration of DAC. The group 0 without DAC was as the control group. Q-MSP was used to detect the state of methylation of the p16 in SCC-9 cells treated by DAC after 48 hours. Real-time fluorescence quantitative PCR was used to detect mRNA expression level changing of the p16 in SCC-9 cells treated by DAC. Immunohistochemical method was used to detect the expression of p16 protein. Results The hypermethylation and non-methylated p16 gene in SCC-9 was mixed with the results of Q-MSP. The results of Real-time PCR showed that mRNA expression of p16 in SCC-9 cells which treated by the different concentration of DAC for 48 hours was higher the control group. And difference was statistically significant (P < 0.05). The high expression of p16 protein was found in the experimental group with immunohistochemical method. Conclusion The p16 gene methylation states of SCC-9 may be suppresses and the recovery of mRNA and protein expression of p16 gene must be prompted by DAC.
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DNA from 32 cases of tongue squamous cell carcinoma(TSCC)specimens and the neck lymph node metastasis specimens were processed by bisulfite treatment.The methylation of the specimens was examined by Q-MSP amplification.The consistency of p1 6 methyla-tion of primary TSCC with that of lymph node metastasis tumor was 90.62%(P <0.05).We assumed that p1 6 gene promoter CpG island methylation rates in primary TSCC and metastatic lymph nodes are consistent,p1 6 gene methylation status can be a methylation predictor be-tween premary and lymph node metstasized TSCC.
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<p><b>OBJECTIVE</b>To investigate the promotive effect of transplantation of bone morphogenetic protein-2 (BMP-2) gene transfected bone mesenchymal stem cells (BMSCs) compounded with injectable bone tissue engineering scaffold material Pluronic F-127 on bone regeneration in rabbit mandibular distraction osteogenesis(DO).</p><p><b>METHODS</b>Forty-eight New Zealand's white rabbits were randomized into four groups with twelve in each. All the objects were prepared into DO surgical model. On the 2nd day of consolidation, group A, B, C and D were injected with the same amount of 200 microL of the compound of BMP-2 gene transfected BMSCs with Pluronic F-127, the solution with BMP-2 gene transfected BMSCs, the solution of BMSCs and physiological saline at distraction zone, respectively. Two halves of the objects of all groups were sacrificed at the end of 2nd and 6th week consolidation, respectively. And the specimens of right mandible were prepared for radiological, histomorphological and immunohistochemical examinations to evaluate bone regeneration.</p><p><b>RESULTS</b>Both radiological and immunohistochemical images were analyzed and processed with professional software. At the end of 2nd and 6th week consolidation, the bone mineral density and the expression of BMP-2 protein in distraction area of group A were significantly higher than those of B, C, D group (P<0.01). Group B was significantly higher than that in group C and D (P<0.01). There was no significant difference between group C and D (P>0.05). And the regeneration quality of distraction zone in group A and B were better than those in group C and D, just as that of group A better than group B. Conclusion The transplantation of BMP-2 gene transfected BMSCs compound with Pluronic F-127 could effectively promote bone regeneration in rabbit mandibular DO.</p>