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目的研究Hedgehog(Hh)信号通路阻断剂环王巴明对前列腺癌Dul45细胞增殖及凋亡的影响。方法体外培养DU145细胞,以10、50、100μmol/L的环王巴明作用24 h,MTT法检测环王巴明对DU145细胞增殖的影响,流式细胞技术检测环王巴明对DU145细胞凋亡的影响,RT-PCR检测环王巴明对DU145细胞Gli-1 m RNA表达的变化,Westernblot检测环王巴明对DU145细胞Gli-1蛋白表达的变化。结果 MTT检测结果表明10μmol/L环王巴明组(85.01±2.61)%、50μmol/L环王巴明组(76.71±3.13)%和100μmol/L环王巴明组(69.10±4.11)%细胞活性较空白对照组(99.97±0.21)%降低(P<0.05);流氏细胞技术检测结果表明10μmol/L环王巴明组(15.12±0.21)%、50μmol/L环王巴明组(24.97±0.24)%和100μmol/L环王巴明组(31.26±0.26)%细胞总凋亡率较空白对照组(2.56±0.28)%升高(P<0.05);RT-PCR检测结果表明10μmol/L环王巴明组(0.69±0.06)%、50μmol/L环王巴明组(0.69±0.06)%和100μmol/L环王巴明组(0.69±0.06)%Gli-1m RNA表达水平较空白对照组(0.92±0.11)%降低(P<0.05);Western blot检测结果表明10μmol/L环王巴明组(0.79±0.06)%、50μmol/L环王巴明组(0.59±0.05)%和100μmol/L环王巴明组(0.41±0.04)%Gli-1蛋白表达水平较空白对照组(0.94±0.11)%降低(P<0.05)。结论 Hedgehog信号通路阻断剂环王巴明可以抑制DU145细胞增殖、诱导DU145细胞凋亡,其作用机制可能与下调GLi-1表达有关。
Objective To investigate the effect of Hedgehog (Hh) signaling pathway inhibitor Wangbamin on the proliferation and apoptosis of prostate cancer Dul45 cells. Methods DU145 cells were cultured in vitro, and treated with 10, 50 and 100μmol / L of eupramine for 24 hours. The proliferation of DU145 cells was detected by MTT assay. The apoptosis of DU145 cells was detected by flow cytometry The changes of Gli-1 mRNA expression in DU145 cells were detected by RT-PCR. The expression of Gli-1 protein in DU145 cells was detected by Western blot. Results The results of MTT showed that the apoptosis rate was significantly increased in 10 μmol / L group (85.01 ± 2.61%), 50 μmol / L group (76.71 ± 3.13%) and 100 μmol / L group (99.97 ± 0.21)% (P <0.05). The results of flow cytometry showed that the inhibitory effect of 10 μmol / L cyclopentamine (15.12 ± 0.21%), 50 μmol / L cyclopentamine (24.97% (P <0.05). The results of RT-PCR showed that the apoptosis rate of 10μmol / L group was significantly higher than that of the blank control group (P <0.05) (0.69 ± 0.06)% in L-leucocytosis group, 0.69 ± 0.06% in 50μmol / L cyclo-amidline group, and 0.69 ± 0.06% (0.92 ± 0.11)% in the control group (0.92 ± 0.11)% (P <0.05). The results of Western blot showed that 10μmol / L omepamine group (0.79 ± 0.06)% and 50μmol / The level of Gli-1 protein expression in the 100 μmol / L Wangbaming group was significantly lower than that in the blank control group (0.94 ± 0.11)% (P <0.05). Conclusion Hedgehog signaling pathway blocker Wangbaming can inhibit the proliferation of DU145 cells and induce the apoptosis of DU145 cells. The mechanism may be related to the down-regulation of GLi-1 expression.