摘要
目的:探讨AKR1C3基因沉默对前列腺癌PC3细胞增殖与转移以及多西紫杉醇药物敏感性的影响。方法:通过脂质体转染的方法将GAPDH-pGIPZ shRNA和AKR1C3-pGIPZ shRNA瞬时转染前列腺癌PC3细胞。实验分为对照组、阴性对照组(GAPDH-pGIPZ shRNA)和实验组(AKR1C3-pGIPZ shRNA)。通过蛋白质印迹法检测转染72h后PC3细胞中AKR1C3蛋白的表达;细胞计数法检测各组前列腺癌PC3细胞的生长曲线及多西紫杉醇对各组细胞的抑制作用;划痕实验检测各组前列腺癌PC3细胞的迁移能力。结果:shRNA转染前列腺癌PC3细胞48h后荧光显微镜下细胞发绿色荧光。蛋白质印迹法结果表明,实验组AKR1C3蛋白表达下降,相对表达量为(5.71±0.03)%,明显低于阴性对照组(9.75±0.08)%和对照组(9.55±0.05)%,F=44.050,P<0.001。同时PC3细胞生长速率变慢,对多西紫杉醇的药物敏感性增加,实验组耐药IC50(27.81±0.02)nmol/L明显低于对照组(60.26±0.03)nmol/L和阴性对照组(59.94±0.05)nmol/L,F=823 200.711,P<0.001。实验组细胞迁移能力下降。结论:前列腺癌PC3中AKR1C3基因的沉默能有效影响肿瘤细胞增殖、迁移和对多西紫杉醇药物的敏感性,提示AKR1C3有望成为前列腺癌治疗的靶基因。
OBJECTIVE：To explore the effect of AKR1C3 gene silencing on prostate cancer PC3 cell proliferation, migration and their sensitivity to docetaxel. METHODS：GAPDH-pGIPZ shRNA and AKR1C3-pGIPZ shRNA were trans fected into PC3 prostate cancer cells by liposome. The experiment was divided into control,negative control（GAPDH-pGI- PZ shRNA） and experimental groups （AKR1C3-pGIPZ shRNA）. Western Blotting was used to detect AKR1C3 protein expression after transfection and cultivation for 72 h in PC3 cells in order to confirm the silence effect. Cell growth curve and the effects of docetaxel on cell proliferation were detected by the cell count method. Cell ability of migration was detec- ted through scratch test. RESULTS： The cells with green fluorescence were found under fluorescence microscope 48 hours after shRNA transfection. Western Blotting results showed that experimental group was decreased in AKR1C3 protein ex-pression, the relative expresssion level was （5.71 ± 0.03 ）%, much lower than that of the negative control group（9.75± 0.08） % and the control group （9.55 ± 0.05 ）% （F= 44. 050, P〈0. 001）. The cell growth was slow down, their ability of migration was inhibited and the sensitivity to docetaxel was increased in experimental group. The ICs0 of experimental group was （27.81±0.02） nmol/L,much lower than that of the control group （60.26±0.03） nmol/L and the negative control （ 59.94 ±0.05 ） nmol/L （F： 823 200.711, P〈0.001 ）. CONCLUSIONS： The silencing of AKR1 C3 gene in PC3 can inhibit their proliferation, migration and the sensitivity to docetaxel. That indicates that AKR1C3 may be one target gene in prostate cancer treatment.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第4期265-268,279,共5页
Chinese Journal of Cancer Prevention and Treatment
