Development of new therapy for hormone independent prostate cancer.

本实验研究了FGFR 2 IIIb在激素非依赖性前列腺癌细胞增殖和分化中的作用，并研究了FGFR 2 IIIb转染细胞在放疗和抗癌药物联合治疗中的作用。1）FGFR 2 IIIb转染PC-3细胞的建立：FGFR 2 IIIb转染PC-3细胞的生长速度明显低于对照细胞。PC-3 FGFR 2 IIIb来源的肿瘤的生长速率显著低于对照。PC-3 FGFR 2 IIIb细胞从PC-3 neo细胞形态上改变。免疫细胞化学分析显示，对照组的pancytokeratin和lactoferin染色呈弱阳性，而PC-3 FGFR 2 IIIb细胞的pancytokeratin和lactoferin免疫染色呈强阳性。2）FGFR 2 IIIb的信号转导：FRS 2在FGF-1刺激的PC-3 neo和PC-3 FGFR 2 IIIb细胞中均被鉴定为酪氨酸磷酸化蛋白。FRS 2在PC-3 FGFR 2 IIIb中的信号强度明显强于对照组。在FGF-7的刺激下，FRS 2在PC-3 v细胞中被强烈激活，但在PC-3 neo细胞中没有。在用FGF-1和FGF-7刺激时，在PC-3 FGFR 2 IIIb细胞中检测到p44/42 MAP激酶的磷酸化，但在PC-3 neo细胞中未检测到。3）放射和抗癌剂与FGFR 2 IIIb的协同作用4和8戈伊的放射处理分别使PC 3-IIIb细胞中的集落数显著减少87.7%和99.6%。携带空载体的Mock细胞中的集落数分别减少了69.4%和95.2%。第3天，不含多西他赛的PC-IIIb的平均细胞活力为0.34，PC 3-IIIb 1 +0.01μM多西他赛的平均细胞活力为0.24，PC 3-IIIb + 0.1μM多西他赛的平均细胞活力为0.20。组间差异具有统计学显著性（p<0.0001）。

The role of FGFR2IIIb in cell proliferation anddifferentiation were studied in hormone independentprostate cancer, and usefulness of combination therapyusing radiation and anticancer agent were also studied in the FGFR2IIIb transfected cell.1)Establishment of FGFR2IIIb transfected PC-3 cell :The growth rate of PC-3 FGFR2IIIb transfected cells was significantly slower than that of the control cells. The growth rate of the PC-3 FGFR2IIIb derived tumors was significantly slower than that of the control. PC-3 FGFR2IIIb cells were morphologically changed from PC-3 neo cells. Immunocytochemical analysis revealed a weak pancytokeratin and lactoferrin-positive staining of the control, whereas PC-3 FGFR2IIIb cells showed strong positive pancytokeratin and lactoferin immunostaining. In the apoptosis assay, most of PC-3 FGFR2IIIb cells were stained with the APOPercentage-dye.2)Signal transduction of FGFR2IIIb :FRS2 was identified as a tyrosine-phosphorylated protein in both PC-3 neo and PC-3 FGFR2IIIb cells stimulated with FGF-1. The signal intensity of FRS2 in PC-3 FGFR2IIIb was much stronger than that of the control. On the stimulation with FGF-7, FRS2 was strongly activated in PC-3 v cells but not in PC-3 neo cells. On stimulation with FGF-1 and FGF-7, the phosphorylation of p44/42 MAP kinase was detected in PC-3 FGFR2IIIb cells, but not in PC-3 neo cells.3)Synergistic effects of radiation and an anticancer agent with the FGFR2IIIbFour and 8 Gy of radiation treatment dramatically decreased the number of colonies in PC3-IIIb cells by 87.7 % and 99.6 %, respectively. The number of colonies in Mock cells bearing an empty vector was reduced by 69.4% and 95.2%, respectively. The mean cell viability on day 3 was 0.34 for PC-IIIb without docetaxel, 0.24 for PC3-IIIb1+0.01μM docetaxel, and 0.20 for PC3-IIIb + 0.1μM of docetaxel. The difference between the groups was statistically significant (p<0.0001).