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BACKGROUND:Neural stem cel s have multi-directional differentiation potential, self-sustaining and self-renewal capacity as wel as have strong migration ability. Bushen Huoxue Recipe can reduce neuronal damage and promote nerve cel regeneration, to achieve neural function reconstruction. Underlying mechanisms of Bushen Huoxue Recipe combined with neural stem cel transplantation in rats with tinnitus induced by sodium salicylate are yet unclear. OBJECTIVE:To investigate the effects of Bushen Huoxue Recipe combined with neural stem cel transplantation in rats with tinnitus induced by sodium salicylate. METHODS:Sixty Sprague-Dawley rats were randomized into five groups (n=12):normal control group, tinnitus model group, Bushen Huoxue Recipe group, stem cel group and combined treatment group (Bushen Huoxue Recipe combined with neural stem cel transplantation). Animal models of tinnitus induced by sodium salicylate were made in al the groups except for the normal control group. Fifteen days after modeling, rats were given intragastric administration of Bushen Huoxue Recipe water decoction (3 mL, 2.592 g/mL) for consecutive 7 days in the Bushen Huoxue Recipe group, intravenous injection of neural stem cel s (1 mL, 1.0×109/L) in the stem cel group, or their combined treatment in the combined treatment group. RESULTS AND CONCLUSION:Bushen Huoxue Recipe, neural stem cel transplantation and their combination al could effectively promote the recovery of drinking water inhibitory rate that was ranked as fol ows:combined treatment group
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OBJECTIVE@#To show that Stat3 played a key role in the G1 to S phase transition in laryngocarcinoma cells.@*METHOD@#Human laryngocarcinoma cell lines Hep-2 were transfected with Stat3 antisense oligonucleotide mediated by liposome, MTT assay was used to measure the proliferation, flow cytometry was applied to analyze the cell cycle, and the expressions of Stat3, phosphorylation specific Stat3 (tyrosine705), CyclinD1, Cyclin E, CDK2, CDK4, CDK6, p21 and p27 were detected by western blot.@*RESULT@#Hep-2 laryngocarcinoma cell lines expressed constitutively activated Stat3. Antisense oligonucleotide which directed blocked up the translation site resulted in growth inhibition, downregulation of Stat3, p-Stat3, Cyclins and CDKs, and upregulation of p21 and p27.@*CONCLUSION@#Our findings suggested that Stat3 played an important role in the G1 to S phase transition in laryngocarcinoma cells, Stat3 orchestrated cell cycle by regulating the balance between CDK/Cyclin complex and CKI.
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Humans , Cell Line, Tumor , G1 Phase , Laryngeal Neoplasms , Metabolism , Pathology , S Phase , STAT3 Transcription Factor , Genetics , Metabolism , Signal Transduction , TransfectionABSTRACT
OBJECTIVE To construct Hep-2 cell line with stable transfection of siRNA-STAT3 gene, and to explore the relationship between STAT3 expressionand the pathogenesis of laryngeal cancer. METHODS Specific STAT3 oligonucleotides were designed and synthesized. These oligonucleotides were annealed form the double strands DNA fragments. Then the fragments were cloned into pGPU6/GFP/Neo vector. The recombinant PGPU6/GFP/Neo-siRNA-STAT3 plasmid was confirmed by enzyme digestion and sequence analysis at the same time. Positive clones were selected out by G418 and p-STAT3 expression was identified by Western-blot analysis. The growth curve of the Hep-2 cells was drawn based on MTT assay and the growth ability of single Hep-2 cell was measured according to the plate colony formation assay respectively. RESULTS PGPU6/GFP/NEO-siRNA-STAT3 expression vector was successfully constructed. Western-blot analysis identified that p-STAT3 expression declined obviously in siRNA- STAT3 group compared with that in negative control and blank group. The growth curve explained that the Hep-2 cells of siRNA-STAT3 group began to show obvious inhibitory effect until they were inoculated in the culture plate for 3 days(P =0.001).There were pronounced inhibitory effect after 5 days(P=0.000). The results also showed time-effect relationship of the inhibition. Furthermore, the cell colony formation rate of siRNA- STAT3 group was less than the negative control and blank group(P =0.000). CONCLUSION We successfully selected out Hep-2 cell line expressed siRNA-STAT3 gene stably and efficiently in this study. Down regulation of STAT3 correlated with the inhibition of growth of Hep-2 cells.
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OBJECTIVE This study was conducted to investigate the expression of Stat3 and its target gene products including Cyclin D1 and Bcl-xl in human laryngeal cancerous tissue, and to explore the mechanism in tumorigenesis of larymgeal carcinoma, and to explore the correlation between the expression,clinical parameters as well as the pathological parameters of laryngeal carcinoma. METHODS The expression of Stat3, p-Stat3, Cyclin D1 and Bcl- xl in 50 cases of cancerous tissues, adjacent normal tissues was measured by Western blot analysis. The expression pattern of Stat3 and its activated form p-Stat3 was determined by immunohisto- chemical staining. The correlation of the expression of Stat3, p-Stat3, Cyclin D1 and Bcl-xl in laryngeal carcinoma with various clinicopathological characteristics was analyzed statistically. RESULTS The protein expression level of stat3, p-Stat3, Cyclin D1, Bcl-xl (A value)were 86.97?1.58, 56.97?3.93, 24.43?5.62, 33.78? 3.56 in laryngeal carcinoma and 36.10?0.52, 16.52?0.52, 5.94?1.75, 14.68?2.14 in adjacent normal respectively(P