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Objective To investigate the effect of lentivirus carrying shRNA-VDR vector on GLi1 in pros-tate cancer PC-3 cells. Methods The cells were cultured according to the culture conditions of PC-3 cells. Expression of VDR and GLi1 in PC-3 cells was detected by fluorescence quantitative PCR and immunocytochemistry SP method.The efficiency of PC-3 cell virus infection was evaluated.The effect of VDR gene interference and GLi1 transcription level on PC-3 cells was detected by RT-PCR.Results Cell culture,cell status was recorded and PC-3 cells were in good condition and the passages was 4 days. Fluorescence quantitative and immunocytochemi-cal SP showed that VDR and GLi1 were expressed in PC-3 cells.The virus infection efficiency showed that the in-fection efficiency was about 95% when adding LV3-NC lentivirus to PC-3 cells at 1:10 ratio. RT-PCR showed that VDR-shRNA lentivirus successfully disturbed VDR expression and decreased by 85%(P < 0.05)compared with the control group after 72 days of VDR-shRNA lentivirus transfection. Transcription level of GLi1 gene in the experimental group increased by 9% compared with the control group(P < 0.05). The transcription level of GLi1 gene in the experimental group increased by 248% compared with the control group(P < 0.05). Conclusion The cultured PC-3 cells were in good condition. VDR and GLi1 genes were expressed in PC-3 cells. Lentivirus showed highest efficiency in infecting PC-3 at 1:10 ratio. When VDR was disturbed,the expression of GLi1 in-creased.In prostate cancer cells,vitamin D can inhibit the Hh signaling pathway and cause GLi1 expression down expression.
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AIM: To explore whether strontium ranelate ( Sr ) promotes osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) through the Hedgehog/Gli1 signaling pathway.METHODS:BMSCs were isolated from 4-week-old rats by adherent culture and induced to differentiate into osteoblasts .According to the experimental purposes , the cells were exposed to different concentrations of Sr , cyclopamine ( Cy, an inhibitor of Hedgehog receptor ) or Gli1-siR-NA.The expression of Gli1 and Runx2 in the cells was detected by Western blotting .The activity of alkaline phosphatase ( ALP) was measured by the method of colorimetry , and the mineralized nodules were observed under microscope with aliz-arin red staining .RESULTS:Exposure to Sr at concentrations of 0.1 to 5 mmol/L for 7 d markedly increased the expres-sion of Gli1 in the BMSCs , and the increase in Gli1 expression was the most obvious following Sr exposure at concentration of 3 mmol/L.Cy at concentration of 10 μmol/L inhibited Sr-induced up-regulation of Gli1 expression.Transfection of the BMSCs with Gli1-siRNA not only obviously inhibited Sr-induced up-regulation of Gli1 and Runx2 ( a downstream protein of Gli1) expression, but also antagonized Sr-induced enhancement of ALP activity and the formation of mineralized nodules . CONCLUSION:The Hedgehog/Gli1 pathway is involved in Sr-induced osteogenic differentiation of rat BMSCs .
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Objective This study analyzes the expression and clinical significance of Gli1 and Gli3 proteins in hepatocellular carcinoma.Methods 36 patients with hepatocellular carcinoma were studied.The expressions of Gli1 and Gli3 in the carcinoma and adjacent non-tumor tissues were detected with immunohistochemical assay,and their correlations with clinicopathological factors were statistically analyzed.Results Expression rates of Gli1 in hepatocellular carcinoma and adjacent nontumor tissues were 75 ％ and 36.1％,respectively.Expression rates of Gli3 in hepatocellular carcinoma and adjacent non-tumor tissues were 58.3％ and 30.6％,respectively.Expression rates of Gli1 and Gli3 in hepatocellular carcinoma were significantly higher than in adjacent non-tumor tissues (P＜0.05),and a positive correlation was found between the expression of Gli1 and Gli3 (r=0.423,P＜0.05).There was no association between the expression of Gli3 and clinicopathological factors such as age,tumor size,tumor differentiation and lymphatic metastasis.The expression of Gll1 was not related witha patient's age and tumor size,hut there were significant associations with tumor differentiation and lymphatic metastasis.Conclusions Therefore,the expression rate of Gli1 was positively correlated with tumor malignancy,which makes the detection of Gli1 and Gli3 valuable for the diagnosis and prognosis of hepatocellular carcinoma.