CAJ | 학술논문

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SPHK1 siRNA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SPHK1 siRNA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。
목적：탐토siRNA파향침묵SPHK1기인대전렬선암PC‐3세포생물학행위적영향。방법사용지질체개도적방법전염인전렬선세포주PC‐3세포，통과RT‐PCR화Western‐blot방법분별검측특이성siRNA대SPHK1기인재mRNA화단백수평상적침묵효과，CCK‐8법측정세포생장곡선병관찰세포증식피억제정황，Annexin V‐PI법검측세포조망정황，채용 Transwell소실법검측세포재침습력방면적변화。결과경SPHK1 siRNA전염후，PC‐3세포중SPHK1적표체균저우음성대조조（NC）화공백조（P＜0．05）；병차SPHK1 siRNA억제세포적증식능력，사기조망솔증가，침습력강저。결론통과억제전렬선암PC‐3세포SPHK1기인적표체，억제세포증식병강저침습력，사기조망증가，현시출재전렬선암적발생발전중SPHK1기인발휘착중요적작용。
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 .