全反式维甲酸联合多西紫杉醇对雄激素非依赖性前列腺癌细胞株PC-3的作用

发布时间：2018-06-08 11:10

本文选题：全反式维甲酸 + 多西紫杉醇　；参考：《昆明医科大学》2014年硕士论文

【摘要】：目的：探讨ATRA联合DOC对雄激素非依赖性前列腺癌细胞株PC-3的作用。方法：将体外培养的前列腺癌PC-3细胞分为对照组(仅加培养基)、ATRA组(10-5mol/L ATRA处理)、DOC组(10-6mol/L DOC处理)及联合组(10-5mol/L ATRA+10-6mol/L DOC处理),作用24h。利用比色法(MTS法)测定细胞增殖抑制情况；普通光学显微镜观察细胞形态学的变化；流式细胞仪测定细胞周期时相变化及检测细胞凋亡；流式细胞仪检测CD133+CD44+细胞比例。结果： ATRA和DOC均可抑制前列腺癌PC-3细胞增殖,10-5、10-6、10-7mol/L ATRA作用24h后对PC-3细胞抑制率分别为37.7%、32.9%、28.2%,10-6、10-7、10-8mol/L DOC作用24h后对PC-3细胞抑制率分别为43.7%、36.2%、32.8%,呈剂量依赖效应,联合用药组作用24h后的细胞抑制率为59.3%,明显高于单独用药组(P0.05),提示联合用药后对前列腺癌PC-3细胞的抑制作用高于单独用药；普通光学显微镜观察到ATRA和DOC均导致细胞皱缩、核质浓缩、核碎裂等凋亡形态学改变,联合用药组表现更明显；流式细胞仪检测显示10-5mol/L ATRA作用24h后PC-3的细胞凋亡率为10.05%,10-6mol/L DOC作用24h后PC-3的细胞凋亡率为12.51%,联合用药组作用24h后PC-3的细胞凋亡率为19.38%,联合用药组显著高于单独用药组(P0.05),提示联合用药对前列腺癌PC-3细胞的促凋亡作用高于单独用药；ATRA所致的细胞周期阻滞以Go/G1期为主,DOC所致的细胞周期阻滞以G2/M期为主,联合用药后G2/M期比例有所下降,同时Go/G1期比例明显增加,提示联合用药后细胞周期比例变化差异有显著性(P0.05)；流式细胞仪检测显示ATRA处理后PC-3细胞中CD133+CD44+细胞比例为0.2%、较阴性组0.6%下降,DOC处理后PC-3细胞中CD133+CD44+细胞比例为3.3%、较阴性组上升,与ATRA组相比差异显著(P0.05),联合用药组处理后CD133+CD44+细胞比例为0.2%,与ATRA组相比无显著差异(P0.05),提示ATRA可能具有促进前列腺癌干细胞分化的作用。结论： 1.DOC作用于前列腺癌PC-3细胞能导致细胞凋亡,但肿瘤干细胞比例却明显升高。 2.ATRA能够降低前列腺癌PC-3的干细胞比例,提示ATRA可能具有分化前列腺癌干细胞的作用。 3.ATRA联合DOC可协同抑制前列腺癌PC-3细胞的增殖,增强二者的促凋亡作用。
[Abstract]:Objective: to investigate the effect of ATRA combined with DOC on androgen independent prostate cancer cell line PC-3. Methods: PC-3 cells cultured in vitro were divided into control group (10-5 mol / L ATRA treated with 10-5 mol / L ATRA) and PC-3 cells treated with 10-6 mol / L DOC in ATRA group; The combined group treated with 10-5 mol / L ATRA 10-6 mol / L DOC for 24 h. Cell proliferation inhibition was measured by colorimetric assay (MTS), morphological changes were observed by ordinary optical microscope, phase changes and apoptosis were detected by flow cytometry (FCM). Results: both ATRA and DOC could inhibit the proliferation of PC-3 cells by flow cytometry. The inhibitory rates of ATRA and DOC on PC-3 cells were 37.7% and 32.7 mol / L ATRA for 24 h, respectively. The inhibitory rates of ATRA and DOC on PC-3 cells were 37.7% and 36.2mol / L, respectively, in a dose-dependent manner. The inhibition rate of PC-3 cells in combined treatment group was 59.3, which was significantly higher than that in single drug group, suggesting that the inhibitory effect of combined treatment on prostate cancer PC-3 cells was higher than that of PC-3 cells treated alone, and ATRA and DOC both caused cell shrinkage under general optical microscope. The morphological changes of apoptosis, such as nuclear cytoplasm concentration and nuclear fragmentation, were more obvious in the combination group. Flow cytometry analysis showed that the apoptosis rate of PC-3 was 10.05% after 24 h exposure to 10-5 mol / L ATRA, and 12.51% of PC-3 cells were treated with 10-6 mol / L DOC for 24 h. The apoptosis rate of PC-3 in combination group was 19.38 after 24 hours of treatment. The results showed that the apoptotic effect of combined therapy on prostate cancer PC-3 cells was higher than that induced by ATRA alone. The cell cycle arrest induced by G / G _ 1 phase and DOC was mainly G _ 2 / M phase. The proportion of G _ 2 / M phase decreased after combined medication, and the proportion of Go-R / G _ 1 phase increased significantly. The results of flow cytometry showed that the percentage of CD133 CD44 cells in PC-3 cells treated with ATRA was 0.2, and the percentage of CD133 CD44 cells in PC-3 cells treated with DOC decreased by 0.6% compared with negative group, which was higher than that in negative group. Compared with ATRA group, the ratio of CD133 CD44 cells was 0.25.There was no significant difference between ATRA group and ATRA group, suggesting that ATRA could promote the differentiation of prostate cancer stem cells. Conclusion: 1. DOC acts on prostate cancer PC-3. Cells can cause apoptosis. However, the percentage of tumor stem cells was significantly increased. 2. ATRA could reduce the percentage of PC-3 cells, suggesting that ATRA might have the function of differentiating prostate cancer stem cells. 3. ATRA combined with DOC could inhibit the proliferation of prostate cancer PC-3 cells. Enhance the apoptotic effect of both.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

【参考文献】