CAJ | 학술논문

目的观察精子相关抗原9(SPAG9)对前列腺癌细胞迁移、侵袭能力的影响.方法采用实时荧光定量聚合酶链反应(RT-qPCR)从mRNA水平评估SPAG9在20例前列腺癌与相应的癌旁组织及在人正常前列腺上皮细胞和前列腺癌细胞株中的表达.应用RNA干扰技术下调SPAG9在前列腺癌细胞株PC-3中的表达,采用RT-qPCR和Western blot方法评估SPAG9基因沉默效果.采用细胞划痕实验和Transwell侵袭实验比较SPAG9基因沉默前后细胞迁移和侵袭能力的改变.结果 80％(16/20)的前列腺癌中SPAG9的表达显著高于相应癌旁组织,且SAPG9的表达与Gleason评分(P＜0.05)和TNM分期(P＜0.05)密切相关.SPAG9基因在3种细胞株中从低到高依次为:人正常前列腺上皮细胞RWPE-1、低侵袭性前列腺癌细胞株LNCaP、高侵袭性前列腺癌细胞株PC-3,前两者与后者比较差异均有统计学意义(P＜0.01).特异性SPAG9基因的小干扰RNA(siRNA)能够有效沉默PC-3细胞中SPAG9 mRNA和蛋白的表达,同时在降低PC-3细胞中SPAG9表达后,发现细胞的迁移[(41.19 ±7.50) μm比(10.39±1.75) μm,P＜0.05]和侵袭[(192.50±26.00)个比(98.65±13.90)个,P＜0.05]能力均显著降低.结论 SPAG9基因表达在前列腺癌细胞迁移和侵袭中发挥重要作用.
목적 관찰정자상관항원9(SPAG9)대전렬선암세포천이、침습능력적영향.방법 채용실시형광정량취합매련반응(RT-qPCR)종mRNA수평평고SPAG9재20례전렬선암여상응적암방조직급재인정상전렬선상피세포화전렬선암세포주중적표체.응용RNA간우기술하조SPAG9재전렬선암세포주PC-3중적표체,채용RT-qPCR화Western blot방법평고SPAG9기인침묵효과.채용세포화흔실험화Transwell침습실험비교SPAG9기인침묵전후세포천이화침습능력적개변.결과 80％(16/20)적전렬선암중SPAG9적표체현저고우상응암방조직,차SAPG9적표체여Gleason평분(P＜0.05)화TNM분기(P＜0.05)밀절상관.SPAG9기인재3충세포주중종저도고의차위:인정상전렬선상피세포RWPE-1、저침습성전렬선암세포주LNCaP、고침습성전렬선암세포주PC-3,전량자여후자비교차이균유통계학의의(P＜0.01).특이성SPAG9기인적소간우RNA(siRNA)능구유효침묵PC-3세포중SPAG9 mRNA화단백적표체,동시재강저PC-3세포중SPAG9표체후,발현세포적천이[(41.19 ±7.50) μm비(10.39±1.75) μm,P＜0.05]화침습[(192.50±26.00)개비(98.65±13.90)개,P＜0.05]능력균현저강저.결론 SPAG9기인표체재전렬선암세포천이화침습중발휘중요작용.
Objective To investigate the effects of sperm associated antigen 9 (SPAG9) gene expression on migration and invasion of human prostate cancer cells.Methods SPAG9 expression was assessed using quantitative real-time polymerase chain reaction (RT-qPCR) in human tumor tissues and the corresponding normal adjacent tissues derived from 20 surgical patients with prostate cancer,and prostate cancer cell lines (PC-3 and LNCaP),normal prostate epithelial cell RWPE-1.PC-3 cells were transfected with specifically SPAG9-targeted small interfering RNA (siRNA),and the silencing efficiency was evaluated using RT-qPCR and western blotting assay.Then compared with scrambled siRNA group,the abilities of migration and invasion of silencing PC-3 cell were estimated by wound-healing model and matrigel-transwell chamber.Results 80％ (16/20) of prostate cancer samples had a high-level expression of SPAG9 gene compared with corresponding normal adjacent tissue.Highly aggressive PC-3 cells significantly overexpressed SPAG9 compared with a human normal prostate epithelial cell RWPE-l (P ＜ 0.01),and less aggressive prostate cancer cell LNCaP (P ＜ 0.01).The SPAG9-targeted siRNA could significantly silence mRNA and protein expression of SPAG9 in the PC-3 cell.Knock-down of SPAG9 gene expression significantly decreased abilities of PC-3 cell migration [(41.19 ±7.50) μm vs.(10.39 ± 1.75) μm,P＜ 0.05] and invasion (192.50±26.00vs.98.65 ±13.90,P＜0.01).Conclusion SPAG9 could exhibit important roles in the migration and invasion of prostate cancer cells.