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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 investigate the effect of AKR1C3 gene silencing on the proliferation and metastasis of prostate cancer PC3 cells and the drug sensitivity of docetaxel. METHODS: GAPDH-pGIPZ shRNA and AKR1C3-pGIPZ shRNA were transiently transfected into prostate cancer PC3 cells by liposome transfection. The experiment was divided into control group, negative control group (GAPDH-pGIPZ shRNA) and experimental group (AKR1C3-pGIPZ shRNA). The expression of AKR1C3 protein in PC3 cells was detected by Western blot 72 h after transfection; the growth curve of PC3 cells in each group and the inhibitory effect of docetaxel on the cells were detected by cell counting method; PC3 cell migration ability. Results: shRNA transfected prostate cancer PC3 cells 48h fluorescent green fluorescence under the microscope. The results of Western blotting showed that the expression of AKR1C3 in the experimental group decreased (5.71 ± 0.03)%, significantly lower than that in the negative control group (9.75 ± 0.08)% and control group (9.55 ± 0.05)%, F = 44.050 P <0.001. At the same time, the growth of PC3 cells slowed down and the sensitivity to docetaxel increased. The IC50 of experimental group was significantly lower than that of control group (60.26 ± 0.03) nmol / L and IC50 (59.94 ± 0.05) nmol / L, F = 823 200.711, P <0.001. Experimental group decreased cell migration. Conclusion: The silence of AKR1C3 gene in prostate cancer PC3 can effectively affect tumor cell proliferation, migration and sensitivity to docetaxel, suggesting that AKR1C3 may be a target gene for prostate cancer treatment.