基因沉默多囊蛋白1和水通道蛋白2对多囊肾囊肿衬里上皮细胞增殖及基质分泌的影响  

The effects of polycystic protein 1 and the water channel protein aquaporin 2 gene silencing on the proliferation and matrix secretion in autosomal dominant polycystic kidney disease cyst-lining epithelial cells

作　　者：江德文[1] 周姗姗[1] 许艳芳[1] 潘阳彬[1] 李桂芬[1] 付槟槟[1] 万建新[1] 

机构地区：[1]福建医科大学附属第一医院肾内科,福建福州350005

出　　处：《中华高血压杂志》2015年第1期73-78,共6页Chinese Journal of Hypertension

基　　金：福建省自然科学基金项目(2012J01339)

摘　　要：目的观察沉默多囊蛋白1(PC-1)和水通道蛋白2(AQP-2)的表达对人常染色体显性遗传性多囊肾囊肿衬里上皮细胞(WT9-12)增殖和基质分泌的影响。方法采用RNA干扰技术(RNAi)特异性抑制PC-1和AQP-2的表达,噻唑蓝法检测细胞增殖能力,流式细胞术检测WT9-12细胞凋亡和周期的变化,酶联免疫吸附试验(ELISA)测定分析细胞基质分泌的变化。结果实验表明,下调细胞中PC-1的表达,在转染24、48、72和96h后,对WT9-12细胞增殖的抑制率分别为12.55%,17.79%、24.69%和35.81%;而下调AQP-2表达,转染72和96h后抑制率为6.77%和8.73%。流式细胞术检测发现下调PC-1可使细胞被阻滞在G0/G1期,并诱导细胞凋亡,而下调AQP-2表达对细胞周期和凋亡无明显的影响;ELISA检测发现沉默AQP-2的表达能抑制Ⅰ型胶原的分泌[(5.53±1.72)比(9.04±2.83)μg/L,P<0.05]。结论应用RNAi可通过下调PC-1的表达抑制WT9-12细胞增殖,并诱导细胞凋亡;而下调细胞AQP-2的表达则可抑制细胞基质Ⅰ型胶原的分泌抑制肾囊肿的发生发展。Objective To observe the effects of silencing polycystic protein 1 （PC-l） and the water channel protein aquaporin 2 （AQP-2） by RNA interference （RNAi） on proliferation and matrix secretion in human autosomal domi- nant polycystic kidney disease （ADPKD） cyst-lining epithelial cells （WT9-12）. Methods Small interfere RNA （siRNA） was transfected to suppress the expression of PC-1 and AQP-2. After knocking-down,PC-1 and AQP 2, WT9-12 cells proliferation was measured by MTT assay,cell cycle and the apoptosis rate by flow cytometry and ex- tracellular matrix secretion by enzyme linked immunosorbent assay kit. Results After down-regulating of PC-1 ex- pression in WT9-12, the celt proliferation inhibition rate was 12.55%, 17.79%, 24.69% and 35.81% after 24, 48, 72 and 96 h post-transfection of PC-1 siRNA respectively. In contrast,AQP-2 knockdown showed only slightly in- hibition on cell proliferation, and the inhibition rate was 6.77% and 8.73% after 72 and 96 h post-transfection of AQP-2 siRNA, respectively. Flow cytometry revealed that down-regulating PC-1 expression can induce cells cycle arrest in G0/G1 phase and apoptosis. When compared with the control group,a reduction in AQP-2 expression ex- erted no obvious change on cell cycle and apoptosis, whereas collagen type 1 secretion was significantly inhibited [（5. 53±1.72） vs （9.04±2.83）μg/L,P〈0.05]. Conclusion The inhibition of PC-1 expression by RNAi could prevent cell proliferation and induce cell apoptosis, and likewise the down-regulation of AQP-2 expression could in- hibit the secretion of extracellular matrix secretion.

关 键 词：RNA干扰技术 常染色体显性多囊肾病 多囊蛋白 水通道蛋白 囊肿衬里上皮细胞 

分 类 号：R692[医药卫生—泌尿科学]