siRNA靶向沉默鞘氨醇激酶-1对前列腺癌PC-3细胞生物学行为的影响  

Effects of SPHK1 RNAi on prostate cancer PC-3 cells

作　　者：李想[1] 时湛 马瑞宁[1] 孙晓青[2] 

机构地区：[1]徐州医学院 [2]徐州医学院附属医院,江苏徐州221000

出　　处：《现代泌尿外科杂志》2015年第9期663-667,共5页Journal of Modern Urology

摘　　要：目的探讨siRNA靶向沉默SPHK1基因对前列腺癌PC-3细胞生物学行为的影响。方法使用脂质体介导的方法转染人前列腺细胞株PC-3细胞,通过RT-PCR和Western-blot方法分别检测特异性siRNA对SPHK1基因在mRNA和蛋白水平上的沉默效果,CCK-8法测定细胞生长曲线并观察细胞增殖被抑制情况,Annexin V-PI法检测细胞凋亡情况,采用Transwell小室法检测细胞在侵袭力方面的变化。结果经SPHK1siRNA转染后,PC-3细胞中SPHK1的表达均低于阴性对照组(NC)和空白组(P<0.05);并且SPHK1siRNA抑制细胞的增殖能力,使其凋亡率增加,侵袭力降低。结论通过抑制前列腺癌PC-3细胞SPHK1基因的表达,抑制细胞增殖并降低侵袭力,使其凋亡增加,显示出在前列腺癌的发生发展中SPHK1基因发挥着重要的作用。Objective To investigate the effects of silencing of sphingosine kinase‐1 （SPHK1） gene expression by siRNA on the biological behavior of human prostate cancer PC‐3 cells .Methods Lipofectamine 2000 （Lipo） was used to transfect the siRNA targeting SPHK1 gene into PC‐3 cells .The expression of SPHK1 was examined by reverse transcription‐polymerase chain reaction （RT‐PCR） and Western‐blot .The change of proliferation was detected by CCK‐8 .The apoptosis was investiga‐ted by Annexin V‐PI .The activities of motility and invasion was assessed by Transwell chamber invasion assay in vitro .Results The expression levels of SPHK1 mRNA and protein in the cells transfected with siRNA were significantly lower than in blank group and in negative control （NC） group （P〈0 .05） .The proliferation of PC‐3 cells was remarkable inhibited ,the invasion was decreased ,and apoptosis was increased .Conclusion The inhibition of SPHK1 gene expression by siRNA could inhibit cells from proliferation ,reduce invasion and increase apoptosis ,which suggest that SPHK1 gene plays a key role in the patho‐genesis and development of prostate cancer .

关 键 词：RNA干扰 前列腺癌 鞘氨醇激酶-1 细胞增殖 细胞凋亡 SPHINGOSINE kinase-1 

分 类 号：R737.25[医药卫生—肿瘤]