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目的:观察前列腺干细胞抗原(prostate stem cell antigen,PSCA)在人雄激素非依赖性前列腺癌PC-3M细胞的表达状况以及PSCA特异性单克隆抗体(PSCA-mAb)对PC-3M细胞生长和凋亡的影响。方法:采用细胞免疫化学方法检测PSCA在PC-3M的表达;以0~1.0μg/mL浓度的PSCA-mAb作用PC-3M细胞0~96h,采用细胞生长曲线、四甲基噻唑氮蓝(methyl thiazolyl tetrazolium,MTT)法、琼脂糖凝胶电泳和流式细胞术(flow cytometry,FCM)分析PC-3M细胞生长及凋亡的变化。结果:PSCA在PC-3M细胞膜和细胞质呈阳性表达;0.1μg/mL PSCA-mAb作用时的细胞生长抑制率为(11.3±2.2)%,0.2μg/mL为(27.5±3.4)%,0.4μg/mL为(39.4±5.8)%,0.6μg/mL为(47.7±7.4)%,0.8μg/mL为(69.3±8.2)%,1.0μg/mL为(70.8±9.3)%,细胞凋亡率为0.1μg/mL为(8.7±1.4)%,0.2μg/mL为(12.3±2.8)%,0.4μg/mL为(21.6±3.2)%,0.6μg/mL为(33.6±4.9)%,0.8μg/mL为(41.4±5.8)%,1.0μg/mL为(42.3±6.1)%,均显著高于相应对照组细胞生长抑制率和凋亡率,P均<0.01;PSCA-mAb作用PC-3M细胞24h的生长抑制率为(47.8±6.4)%,48h为(59.4±7.3)%,72h为(70.3±7.9)%,96h为(71.1±9.0)%,细胞凋亡率24h为(33.6±4.3)%,48h为(36.3±5.1)%,72h为(42.7±5.7)%,96h为(43.4±6.3)%,均高于相应的对照组细胞生长抑制率和凋亡率,P均<0.01。MTT及FCM分析结果表明,PSCA-mAb在0.6μg/mL作用72h时,细胞生长抑制率为(71.5±6.2)%,凋亡率为(44.4±5.5)%,均达到最大。结论:PSCA在人雄激素非依赖性前列腺癌PC-3M细胞呈阳性表达;PSCA-mAb能够时间-剂量依赖性地抑制PC-3M生长和诱导其凋亡。
OBJECTIVE: To observe the expression of prostate stem cell antigen (PSCA) in human androgen-independent prostate cancer PC-3M cells and the effect of PSCA-specific monoclonal antibody (PSCA-mAb) on PC-3M cell growth and apoptosis The impact of death. Methods: The expression of PSCA in PC-3M cells was detected by immunocytochemistry. PC-3M cells were treated with PSCA-mAb at concentration of 0 ~ 1.0μg / mL for 0-96h. Cell growth curve, Thiazolyl tetrazolium (MTT) assay, agarose gel electrophoresis and flow cytometry (FCM) were used to analyze the changes of PC-3M cell growth and apoptosis. Results: The PSCA expression was positive in PC-3M cell membrane and cytoplasm. The cell growth inhibition rate was (11.3 ± 2.2)% at 0.1μg / mL PSCA-mAb, (27.5 ± 3.4)% at 0.2μg / mL, / (39.4 ± 5.8)%, 0.6μg / mL (47.7 ± 7.4)%, 0.8μg / mL (69.3 ± 8.2)% and 1.0μg / mL Was (8.7 ± 1.4)% at 0.1 μg / mL, (12.3 ± 2.8)% at 0.2 μg / mL and 21.6 ± 3.2% at 0.4 μg / mL and 33.6 ± 4.9% at 0.8 μg / mL (41.4 ± 5.8)% and 1.0μg / mL (42.3 ± 6.1)%, respectively, which were significantly higher than that of the corresponding control group (P <0.01) The growth inhibition rate of 3M cells for 24 h was (47.8 ± 6.4)%, (48.4 ± 7.3)% for 48 h, (70.3 ± 7.9)% for 72 h, (71.1 ± 9.0)% for 96 h, ± 4.3%, 48h 36.3 ± 5.1%, 72h 42.7 ± 5.7%, and 96h 43.4 ± 6.3%, respectively, which were significantly higher than that of the corresponding control group (P <0.05) <0.01. The results of MTT and FCM showed that the cell growth inhibition rate and the apoptosis rate of PSCA-mAb were (71.5 ± 6.2) and (44.4 ± 5.5)%, respectively, at 0.6μg / mL for 72h. CONCLUSIONS: PSCA is positively expressed in human androgen-independent prostate cancer PC-3M cells. PSCA-mAb inhibits PC-3M growth and induces apoptosis in a time-and dose-dependent manner.