CAJ | 학술논문

目的研究PIWIL2在人类膀胱尿路上皮癌(BTCC)中的表达及siRNA对人类膀胱癌细胞PIWIL2表达的影响.方法采用反转录聚合酶链反应(RT-PCR)检测BTCC 46例、腺性膀胱炎21例、癌旁组织17例、正常膀胱组织14例中PIWIL2mRNA的表达；设计合成针对PIWIL2的3个特异性siRNA,转入人类膀胱癌BIU-87细胞,分别采用四甲基偶氮唑蓝、DNA原位末端标记法、RT-PCR和Western blot检测siRNA对BIU-87细胞生长抑制率(IR)、凋亡指数(AI)和PIWIL2 mRNA及其蛋白表达的影响.结果 BTCC组织中PIWIL2 mRNA的表达率为76.08％(35/40),均显著高于腺性膀胱炎组织[42.86 ％(9/21)]、癌旁组织[41.17％(7/17)]和正常膀胱组织[7.14％(1/14)] (P=0.008,P=0.010,P=0.000).BTCC组织中PIWIL2的高表达与肿瘤淋巴结转移、病理分级均密切相关.siRNA1 ～3组细胞的IR[( 37.52±8.84)％、(64.36±9.64)％、(62.94±8.43)％]和AI[(26.18±5.42)％、(38.75±6.19)％、(40.02±5.64)％]均分别显著高于对照组[(1.97±0.02)％、(3.35±0.47)％](均P=0.000),PIWIL2mRNA及其蛋白表达水平均显著低于对照组；其中siRNA 2、3组细胞的IR、AI和对PIWIL2表达的抑制作用均显著高于siRNA 1组.结论 PIWIL2在BTCC中的高表达,提示其与膀胱癌的发生、发展密切相关；体外转录合成的siRNA可抑制BIU-87细胞PIWIL2的表达,诱导肿瘤细胞凋亡,从而抑制肿瘤细胞生长,提示PIWIL2可作为膀胱肿瘤基因治疗的重要靶点.
목적 연구PIWIL2재인류방광뇨로상피암(BTCC)중적표체급siRNA대인류방광암세포PIWIL2표체적영향.방법 채용반전록취합매련반응(RT-PCR)검측BTCC 46례、선성방광염21례、암방조직17례、정상방광조직14례중PIWIL2mRNA적표체；설계합성침대PIWIL2적3개특이성siRNA,전입인류방광암BIU-87세포,분별채용사갑기우담서람、DNA원위말단표기법、RT-PCR화Western blot검측siRNA대BIU-87세포생장억제솔(IR)、조망지수(AI)화PIWIL2 mRNA급기단백표체적영향.결과 BTCC조직중PIWIL2 mRNA적표체솔위76.08％(35/40),균현저고우선성방광염조직[42.86 ％(9/21)]、암방조직[41.17％(7/17)]화정상방광조직[7.14％(1/14)] (P=0.008,P=0.010,P=0.000).BTCC조직중PIWIL2적고표체여종류림파결전이、병리분급균밀절상관.siRNA1 ～3조세포적IR[( 37.52±8.84)％、(64.36±9.64)％、(62.94±8.43)％]화AI[(26.18±5.42)％、(38.75±6.19)％、(40.02±5.64)％]균분별현저고우대조조[(1.97±0.02)％、(3.35±0.47)％](균P=0.000),PIWIL2mRNA급기단백표체수평균현저저우대조조；기중siRNA 2、3조세포적IR、AI화대PIWIL2표체적억제작용균현저고우siRNA 1조.결론 PIWIL2재BTCC중적고표체,제시기여방광암적발생、발전밀절상관；체외전록합성적siRNA가억제BIU-87세포PIWIL2적표체,유도종류세포조망,종이억제종류세포생장,제시PIWIL2가작위방광종류기인치료적중요파점.
Objective To investigate the gene expression of PIWIL2 in the bladder urothelial carcinoma (BTCC) and siRNA interact on PIWIL2 gene expression in human bladder cancer cell line BIU-87.Methods Semi-quantitative reverse transcription polymerase chain reaction (qRT-PCR) was applied to detect the PIWIL2 expressions in tissues of BTCC (46 cases),cystitis glandularis(21 cases),adjacent non-cancerous tissues (17 cases) and normal bladder tissues (7 cases). 3 specific siRNA targeted PIWIL2 gene were synthesized after designed and transferred. After siRNA was transferred into BIU-87 cells, MTI and TUNEL methods were applied to detect the proliferation inhibitory rate (IR) and apoptosis index (AI) in BIU-87 cells,qRT-PCR and Western blot were used to examine effects of siRNA on the expressions of the PIWIL2 gene and protein,respectively.Results The expression rate of PIWIL2 mRNA in BTCC tissues was 76.08 ％(35/46) and significantly higher than those in the cystitis glandularis tissues (42.86 ％,9/21),adjacent non-cancerous tissues (41.17 ％,7/17) and normal tissues (7.14 ％,1/14) (P =0.008,P =0.010,P =0.000).The IR [(37.52±8.84) ％,(64.36±9.64)％] and (62.94±8.43) ％] and AI [(26.18±5.42) ％,(38.75±6.19) ％ and (40.02±5.64) ％] of BIU-87 cells in the siRNA 1～3 groups were respectively significantly higher than those [(1.97±0.02) ％ and (3.35±0.47) ％] in the control group(P=0.000),and expressions of PIWIL2 mRNA and protein in the siRNA groups were both lower than those in the control group. Moreover, the effects of siRNA 2 group and siRNA 3 group on inhibiting PIWIL2 expression, IR and AI of BIU-87 cells were stronger than siRNA 1 group. Conclusion The over-expression of PIWIL2 suggested that it played an important role in the mechanism of development and malignant progression of BTCC. The siRNA of transcription can significantly inhibit its expression, induce cell apoptosis and inhibit the growth of BIU-87 cells which might provide the experimental evidence for the gene targeting therapy of bladder tumor.