CAJ | 학술논문

目的研究血小板源性生长因子-D(platelet-derived growth factor-D,PDGF-D)对前列腺癌细胞迁移的影响及其作用机制. 方法蛋白质印迹法检测前列腺癌LNCaP、PC-3细胞中PDGF-D的表达.针对PDGF-D基因的mRNA序列合成siRNA,脂质体转染法转染PC-3细胞；外源性PDGF-D作用LNCaP、PC-3细胞.Boyden槽迁移法分析外源性PDGF-D及siRNA转染对前列腺癌LNCaP、PC-3细胞迁移的影响,RT-PCR法检测血管内发生长因子(vascular endothelial growth factor,VEGF)及基质金属蛋白酶-9(matrix metalloproteinase-9,MMPO)mRNA表达水平的变化. 结果 LNCaP和PC-3细胞中PDGF-D蛋白的表达分别为29.47±1.68和63.43±2.10,两者比较差异有统计学意义(P＜0.05)；PC-3细胞转染PDCF-D siRNA后,siRNA阴性组蛋白表达水平为65.03±2.31,siRNA阳性组为35.19±1.51,两组比较差异有统计学意义(P＜0.05).外源性PDGF-D可以促进LNCaP和PC-3细胞的迁移,并刺激PC-3细胞VEGF、MMP-9 mRNA表达水平升高,与对照组比较,差异均有统计学意义(P ＜0.05)；PDGF-D siRNA转染PC-3细胞后,细胞迁移率下降,VEGF、MMP-9 mRNA的表达水平由转染前的0.72±0.09、0.65±0.07下降为0.43±0.18、0.22±0.08,差异有统计学意义(P＜0.05). 结论 PDCF-D能促进前列腺癌的转移和血管的形成.
목적 연구혈소판원성생장인자-D(platelet-derived growth factor-D,PDGF-D)대전렬선암세포천이적영향급기작용궤제. 방법 단백질인적법검측전렬선암LNCaP、PC-3세포중PDGF-D적표체.침대PDGF-D기인적mRNA서렬합성siRNA,지질체전염법전염PC-3세포；외원성PDGF-D작용LNCaP、PC-3세포.Boyden조천이법분석외원성PDGF-D급siRNA전염대전렬선암LNCaP、PC-3세포천이적영향,RT-PCR법검측혈관내발생장인자(vascular endothelial growth factor,VEGF)급기질금속단백매-9(matrix metalloproteinase-9,MMPO)mRNA표체수평적변화. 결과 LNCaP화PC-3세포중PDGF-D단백적표체분별위29.47±1.68화63.43±2.10,량자비교차이유통계학의의(P＜0.05)；PC-3세포전염PDCF-D siRNA후,siRNA음성조단백표체수평위65.03±2.31,siRNA양성조위35.19±1.51,량조비교차이유통계학의의(P＜0.05).외원성PDGF-D가이촉진LNCaP화PC-3세포적천이,병자격PC-3세포VEGF、MMP-9 mRNA표체수평승고,여대조조비교,차이균유통계학의의(P ＜0.05)；PDGF-D siRNA전염PC-3세포후,세포천이솔하강,VEGF、MMP-9 mRNA적표체수평유전염전적0.72±0.09、0.65±0.07하강위0.43±0.18、0.22±0.08,차이유통계학의의(P＜0.05). 결론 PDCF-D능촉진전렬선암적전이화혈관적형성.
Objective To investigate the effect of platelet-derived growth factor-D (PDGF-D) on the prostate cancer cells migration and its possible mechanism. Methods The expressions of PDGF-D in LNCaP and PC-3 cells were detected with western blot.PDGF-D siRNA was synthesized according to mRNA sequence of PDGF-D gene and was transfected into PC-3 cell.The cells were treated with PDGF-D and PDGF-D siRNA,the cell migration was examined by Boyden chamber migration assay.The expression changes of VEGF and MMP-9 mRNA were detected by RT-PCR. Results The results of western blot indicated that the PDGF-D protein expression level was lower in LNCaP cells (29.47 ± 1.68) than that in PC-3 cells (63.43 ±2.10),(P ＜ 0.05).PDGF-D siRNA could down-regulate the PDGF-D protein expression in the transfected group (35.19 ± 1.51).The exogenous PDGF-D could promote migration of LNCaP and PC-3cells,and up-regulate the expression of VEGF,MMP-9 mRNA in PC-3 cells (P ＜ 0.05,compared with control group).PDGF-D siRNA inhibited PC-3 cells' migration and decreased the level of VEGF,MMP-9mRNA expression (0.72 ± 0.09 vs 0.43 ± 0.18,0.65 ±0.07 vs 0.22 ± 0.08) (P ＜ 0.05). Conclusion PDGF-D is involved in the promotion of prostate cancer invasion and angiogenesis.