中华泌尿外科杂志
中華泌尿外科雜誌
중화비뇨외과잡지
CHINESE JOURNAL OF UROLOGY
2010年
7期
467-470
,共4页
郭永灿%罗春丽%蔡晓钟%谢建红%欧俐苹%赵懿%吕纯芳%冀慧莹%吴小候
郭永燦%囉春麗%蔡曉鐘%謝建紅%歐俐蘋%趙懿%呂純芳%冀慧瑩%吳小候
곽영찬%라춘려%채효종%사건홍%구리평%조의%려순방%기혜형%오소후
膀胱肿瘤%癌%RNA干扰%基因
膀胱腫瘤%癌%RNA榦擾%基因
방광종류%암%RNA간우%기인
Bladder neoplasms%Carcinoma%RNAi%Genes
目的 探讨RNA干扰技术沉默PLCε基因表达对膀胱癌BIU-87细胞株增殖的抑制作用.方法 构建靶向PLCε基因特异性shRNA重组质粒pGenesil-PLCE,经脂质体介导转染BIU-87细胞株,蛋白质印迹法、RT-PCR方法检测PLCε在癌细胞蛋白及mRNA水平的表达,噻唑盐法观察重组质粒对BIU-87细胞增殖的抑制作用,免疫细胞化学染色分析增殖细胞核抗原(PCNA)含量改变,流式细胞仪检测细胞周期分布.结果 构建的重组质粒pGenesil-PLCε能明显抑制PLCE基因表达,且能明显抑制BIU-87细胞增殖;转染重组质粒0、24、48和72 h后细胞增殖抑制率分别为2.01%、32.85%、39.78%、37.19%,重组质粒组与对照组、阴性质粒组比较,差异有统计学意义(P<0.01).细胞内PCNA表达下调了33.08%;G0/G1期细胞(71.50±4.48)%与空白对照组(40.75±2.30)%、阴性质粒组(40.00±1.76)%比较明显增多,G2/M期细胞(8.16±0.51)%与空白对照组(31.20±1.76)%、阴性质粒组(35.94±1.58)%相比减少.差异均有统计学意义(P<0.01),细胞阻滞于G0/G1期,细胞增殖状态受到明显抑制.结论 PLCε基因在促进膀胱癌细胞增殖中发挥莺要作用,可能成为膀胱癌生物学治疗潜在的分子靶点.
目的 探討RNA榦擾技術沉默PLCε基因錶達對膀胱癌BIU-87細胞株增殖的抑製作用.方法 構建靶嚮PLCε基因特異性shRNA重組質粒pGenesil-PLCE,經脂質體介導轉染BIU-87細胞株,蛋白質印跡法、RT-PCR方法檢測PLCε在癌細胞蛋白及mRNA水平的錶達,噻唑鹽法觀察重組質粒對BIU-87細胞增殖的抑製作用,免疫細胞化學染色分析增殖細胞覈抗原(PCNA)含量改變,流式細胞儀檢測細胞週期分佈.結果 構建的重組質粒pGenesil-PLCε能明顯抑製PLCE基因錶達,且能明顯抑製BIU-87細胞增殖;轉染重組質粒0、24、48和72 h後細胞增殖抑製率分彆為2.01%、32.85%、39.78%、37.19%,重組質粒組與對照組、陰性質粒組比較,差異有統計學意義(P<0.01).細胞內PCNA錶達下調瞭33.08%;G0/G1期細胞(71.50±4.48)%與空白對照組(40.75±2.30)%、陰性質粒組(40.00±1.76)%比較明顯增多,G2/M期細胞(8.16±0.51)%與空白對照組(31.20±1.76)%、陰性質粒組(35.94±1.58)%相比減少.差異均有統計學意義(P<0.01),細胞阻滯于G0/G1期,細胞增殖狀態受到明顯抑製.結論 PLCε基因在促進膀胱癌細胞增殖中髮揮鶯要作用,可能成為膀胱癌生物學治療潛在的分子靶點.
목적 탐토RNA간우기술침묵PLCε기인표체대방광암BIU-87세포주증식적억제작용.방법 구건파향PLCε기인특이성shRNA중조질립pGenesil-PLCE,경지질체개도전염BIU-87세포주,단백질인적법、RT-PCR방법검측PLCε재암세포단백급mRNA수평적표체,새서염법관찰중조질립대BIU-87세포증식적억제작용,면역세포화학염색분석증식세포핵항원(PCNA)함량개변,류식세포의검측세포주기분포.결과 구건적중조질립pGenesil-PLCε능명현억제PLCE기인표체,차능명현억제BIU-87세포증식;전염중조질립0、24、48화72 h후세포증식억제솔분별위2.01%、32.85%、39.78%、37.19%,중조질립조여대조조、음성질립조비교,차이유통계학의의(P<0.01).세포내PCNA표체하조료33.08%;G0/G1기세포(71.50±4.48)%여공백대조조(40.75±2.30)%、음성질립조(40.00±1.76)%비교명현증다,G2/M기세포(8.16±0.51)%여공백대조조(31.20±1.76)%、음성질립조(35.94±1.58)%상비감소.차이균유통계학의의(P<0.01),세포조체우G0/G1기,세포증식상태수도명현억제.결론 PLCε기인재촉진방광암세포증식중발휘앵요작용,가능성위방광암생물학치료잠재적분자파점.
Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.
