【摘要】：目的探討雷帕霉素抗神經(jīng)膠質(zhì)瘤的腫瘤活性及其相關(guān)作用機制。方法 MTT細胞毒實驗檢測雷帕霉素對神經(jīng)膠質(zhì)瘤細胞C6的生長抑制活性,Annexin V/PI雙染流式細胞儀檢測細胞凋亡的發(fā)生,PI單染流式細胞儀檢測細胞周期的變化,提取細胞總RNA,RT-PCR檢測survivin mRNA的表達變化,提取細胞總蛋白Western印跡檢測survivin蛋白的表達變化。結(jié)果雷帕霉素對神經(jīng)膠質(zhì)瘤細胞C6顯示了高效的抗腫瘤活性,IC50值為(8.34±0.62)μmol/L。流式細胞儀凋亡分析顯示0、5、10、20μmol/L雷帕霉素作用C6細胞48 h后,細胞的凋亡率分別為(3.56%±1.04%)、(13.34%±3.86%)、(22.84%±4.53%)、(51.91%±6.29%),各組間有統(tǒng)計學差異(P0.01)。細胞周期分析顯示0、5、10、20μmol/L雷帕霉素處理C6細胞48h后,細胞增殖指數(shù)分別為(64.54%±6.04%)、(50.24%±4.86%)、(39.26%±5.61%)、(30.08%±6.42%)。RT-PCR和Western印跡檢測顯示雷帕霉素可以下調(diào)survivin mRNA和蛋白的表達。結(jié)論雷帕霉素具有高效的抗神經(jīng)膠質(zhì)瘤活性,其作用機制可能與其抑制了m TOR信號通路下調(diào)survivin靶蛋白的表達有關(guān)。
[Abstract]:Objective to investigate the anti-glioma activity of rapamycin and its mechanism. Methods MTT cytotoxicity assay was used to detect the inhibitory activity of rapamycin on C6 glioma cells, Annexin V/PI double staining flow cytometry was used to detect apoptosis, and PI single staining flow cytometry was used to detect cell cycle changes. Total RNA, was extracted from glioma cells. The expression of survivin mRNA was detected by RT-PCR, and the expression of survivin was detected by Western blot. Results rapamycin showed high antitumor activity against C6 glioma cells, IC _ (50) was (8.34 鹵0.62) 渭 mol/L.. The apoptosis rate of C6 cells treated with 0, 5, 10, 20 渭 mol / L rapamycin for 48 h was 3.56% 鹵1.04%), (13.34% 鹵3.86%), (22.84% 鹵4.53%, respectively, and the apoptosis rate of C6 cells treated with rapamycin was 3.56% 鹵1.04%, 13.34% 鹵3.86%, 22.84% 鹵4.53%, respectively. (51.91% 鹵6.29%), there was statistical difference among the groups (P0.01). Cell cycle analysis showed that the proliferation index of C6 cells treated with 0, 5, 10 and 20 渭 mol / L rapamycin for 48 h was 64.54% 鹵6.04%), (50.24% 鹵4.86% 鹵4.86%), (39.26% 鹵5.61%, respectively. (30.08% 鹵6.42%). RT-PCR and Western blot analysis showed that rapamycin could down-regulate the expression of survivin mRNA and protein. Conclusion rapamycin has high anti-glioma activity and its mechanism may be related to its inhibition of m-TOR signaling pathway down-regulating the expression of survivin target protein.
【作者單位】： 青海省第五人民醫(yī)院;
【分類號】：R739.41

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