【作者】 刘双林；

【导师】 叶章群； 胡志全；

【作者基本信息】 华中科技大学 ， 外科学， 2011， 博士

【摘要】 目的了解姜黄素在体外对雄激素非依赖性前列腺癌细胞的抑制效应。方法采用不同浓度的姜黄素(6.25μmol/L、12.5μmol/L、25μmol/L、50μmol/L和100μmol/L)作用于雄激素非依赖性前列腺癌细胞,MTT法检测姜黄素对细胞生长活性的影响。然后分别采用CFSE染色、Annexin V-PI染色和PI染色法,使用流式细胞仪分析姜黄素对PC-3细胞增殖、凋亡和细胞周期的影响。结果MTT结果显示6.25μmol/L、12.5μmol/L、25μmol/L和50μmol/L四个浓度的姜黄素处理24小时后,PC-3细胞的活性未受显著影响；而100μmol/L的姜黄素则对PC-3细胞产生显著的细胞毒性作用。50μmol/L的姜黄素作用后,PC-3细胞的增殖指数从7.08±0.20降到4.38±0.19(P<0.05)；凋亡率从(5.34±0.96)%提高到(21.53±2.87)%(P<0.05)；G2/M期细胞比例由(34.27±1.87)%上升到(57.29±1.91)%(P<0.05)。结论姜黄素能在体外显著抑制雄激素非依赖性前列腺癌细胞的增殖,诱导细胞凋亡并导致细胞周期的阻滞。目的明确姜黄素对雄激素非依赖性前列腺癌细胞内信号通路活性的影响,探讨姜黄素抗瘤效应产生的可能机制。方法采用荧光素酶报告基因系统检测姜黄素在体外对核因子-кB (NF-кB、激活蛋白-1(AP-1).Elkl.CHOP和CREB五个信号通路的影响,以带有荧光素酶报告基因的质粒(pNF-κB-Luc,pAP-1-Luc,pElk1-Luc,pCHOP-Luc,pCREB-Luc)转染PC-3细胞,姜黄素干预后,检测荧光素酶活性。结果荧光素酶活性检测结果显示,姜黄素能在体外显著抑制NF-кB和AP-1两个细胞内信号通路的活性(P<0.05),而对Elkl、CHOP和CREB三个信号通路的活性影响不明显。结论姜黄素可能通过抑制NF-кB和AP-1信号通路发挥抑癌效应。目的检测抗人转铁蛋白受体单克隆抗体(TfR mAb)对雄激素非依赖性前列腺癌细胞生物学行为的影响,了解抗人TfR mAb对姜黄素抗瘤作用的增强效应。方法利用杂交瘤细胞株制备抗人TfR mAb,分别采用CFSE染色、AnnexinⅤ-PI染色和PI染色法检测抗人TfR mAb对PC-3细胞增殖、凋亡和细胞周期的影响。采用FITC标记的鼠抗人TfR mAb检测姜黄素对雄激素非依赖性前列腺癌细胞表面转铁蛋白受体表达的影响。采用流式细胞仪进一步检测抗人TfR mAb联合姜黄素对PC-3细胞增殖、凋亡和细胞周期的影响。结果抗人TfR mAb对PC-3细胞增殖、凋亡和细胞周期无显著影响(P>0.05)。姜黄素能够显著上调雄激素非依赖性前列腺癌细胞表面TfR表达(P<0.05)。抗人TfRmAb可显著增强姜黄素对PC-3细胞凋亡的诱导作用(P<0.05),而不影响姜黄素对PC-3细胞增殖和细胞周期的影响(P>0.05)。结论姜黄素可导致雄激素非依赖性前列腺癌细胞表面转铁蛋白受体表达增高。抗人TfR mAb单独作用对雄激素非依赖性前列腺癌细胞的生物学行为无显著影响,但抗人TfR mAb可以增强姜黄素对雄激素非依赖性前列腺癌细胞的凋亡效应。更多还原

【Abstract】 Objective:To investigate the anti-tumor activity of curcumin in androgen independent prostate cancer in vitro.Methods:After PC-3 cells were treated with 6.25μmol/L,12.5μmol/L,25μmol/L, 50μmol/L, and 100μmol/L curcumin, the cell activity was determined by MTT assay. After curcumin treatment, the effects of curcumin on the proliferation of prostate cancer cells were assessed by CFSE dilution. Flow cytometery (FCM) were performed to analysis the cell cycle and the induction of apoptosis on tumor cells.Results:After treating with 50μmol/L curcumin, the proliferation index of androgen independent prostate cancer cells in vitro decreased from 7.08±0.20 to 4.38±0.19 (P< 0.05);the percentage of apoptotic cells increased from 5.34%±0.96% to 21.53%±2.87%(P< 0.05);and the percentage of G2/M phase cells increased from 34.27%±1.87% up to 57.29%±1.91%(P<0.05).Conclusion:Curcumin with the concentration of 50μmol/L and below hadn’t obvious effect on cell viability of androgen independent prostate cancer cells (P<0.05). Curcumin could significantly inhibit the androgen independent prostate cancer through blocking cellular proliferation and inducing apoptosis. Objective:To investigate the effect of curcumin on the intracellular signaling pathway activities of androgen independent prostate cancer and clarify the anti-tumor mechanisms of curcunin.Methods:The PathDetect Cis-/Trans- Reporting Systems (pNF-KB-Luc, pAP-1-Luc, pElkl-Luc, pCHOP-Luc, pCREB -Luc) were transiently transfected into PC3 cells, respectively. After 8 h of transfection, cells were treated with 50μmol/L curcumin for 24 h. The activity of each signaling pathway was determined by measuring the luciferase activities of the reporter in treated and untreated transfectants.Results:Curcumin inhibited the activity of NF-κB and AP-1 associated signaling pathway significantly (P< 0.05), but had little effect on the activity of Elkl, CHOP and CREB signaling pathway.Conclusions:Curcumin exerts anti-tumor effects via inhibition of the NF-κB and AP-1 associated signaling pathway. Objective:To study the anti-tumor activity of anti-human transferrin receptor monoclonal antibody (TfR mAb) in androgen independent prostate cancer in vitro and investigate the effect of TfR mAb on the anti-tumor activity of curcumin.Methods:After curcumin treatment, transferrin receptor expression was detected by flow cytometry on androgen-independent prostate cancer cells. And the cell activity of prostate cancer cells after TfR mAb treatment was determined by MTT assay. After treating with TfR mAb and curcumin, the proliferation of prostate cancer cells were assessed by CFSE dilution. Flow cytometery (FCM) were performed to analysis the cell cycle and the induction of apoptosis on tumor cells.Results:Curcumin could increase the transferrin receptor expression of androgen-independent prostate cancer cells significantly (P<0.05). Anti-human TfR mAb alone had not significant effect on the proliferation, apoptosis and cell cycle of androgen-independent prostate cancer cells (P>0.05). But anti-human TfR mAb enhance the inhibitory effect of curcumin on the apoptosis of PC-3 cells significantly (P<0.05).Conclusion:Curcumin treatment could increase the transferrin receptor expression of androgen-independent prostate cancer cells. Anti-human TfR mAb alone had not obvious anti-tumor activity in androgen independent prostate cancer cells. Blocking the transferrin receptor could enhance the apoptosis effect of curcumin on the proliferation of androgen-independent prostate cancer cells.更多还原