沉默肝细胞再生磷酸酶-3对LNCaP细胞增殖及凋亡的影响

Silencing phosphatase of regenerating liver cell-3 gene regulates proliferation and apoptosis of LNCaP cells

目的观察小干扰RNA（siRNA）沉默促肝细胞再生磷酸酶-3（PRL-3）基因对人雄激素依赖性前列腺癌（LNCaP）细胞增殖及凋亡的影响。方法应用靶向PRL-3基因小干扰RNA转染处理前列腺癌LNCaP细胞后，采用逆转录-聚合酶链反应（RT—PCR）和Westernblot检测PRL-3mRNA和蛋白水平，采用细胞计数试剂盒（CCK-8）检测细胞体外增殖能力，采用流式细胞仪检测细胞凋亡水平。结果与对照组比较，siRNA转染组PRL．3mRNA和蛋白水平明显下调，且呈浓度依赖性（P〈0．05）。体外实验显示，低浓度PRL-3siRNA对LNCaP细胞体外增殖能力没有影响，而中、高浓度PRL-3siRNA转染48h后明显地抑制LNCaP细胞的体外增殖能力（P〈0．05），低、中、高浓度siRNA沉默PRL-3基因后体外LNCaP细胞的凋亡率分别为（28．1±3．8）％、（25．2±2．5）％、（27．1±0．7）％（P〈0．05）。结论下调PRL-3基因的表达可抑制LNCaP细胞的增殖并促进LNCaP细胞的凋亡，PRL-3基因在前列腺癌LNCaP细胞体外增殖及凋亡中起重要作用。

Objective To study the effects of small interfering RNA （siRNA） silencing phosphatase of regenerating liver cell 3 （PRL-3） on proliferation and apoptosis of human prostate cancer LNCaP cells. Methods After prostate cancer LNCaP cells were transfected by PRL-3 siRNA, the mRNA and protein of PRL-3 were detected by reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting, respectively. The growth of LNCaP cells was exmined by cell counting kit-8 （CCK-8）. Flow eytometry was used to detect apoptosis. Results The siRNA could downregulate the mRNA and protein expression level of PRL-3 in a dose-dependent manner （ P 〈 0. 05）. Low doses of PRL-3 siRNA did not affect the proliferation of LNCaP cells, but 100 nmol/L PRL-3 siRNA could effectively inhibit the proliferation of LNCaP ceils after 48 h in vitro （ P 〈 0. 05 ）. Apoptosis rate of LNCaP ceils with low, middle and high PRL-3 siRNA interventions in vitro was （28.1 ± 3.8 ） %, （ 25.2 ± 2. 5 ） %, and （27. 1 ± 0. 7 ） % respectively （ P 〈 0. 05 ）. Conclusion Downregulating PRL-3 could inhibit proliferation of LNCaP cells and promote apoptosis of LNCaP cells. PRL-3 gene might play an important role in proliferation and apoptosis of LNCaP cells.