CAJ | 학술논문

目的探讨去甲基化药物5-氮-2'-脱氧胞苷(DAC)对膀胱肿瘤细胞生长的抑制作用.方法利用细胞增殖试剂WST-1法测定不同浓度DAC对RT112、253J、T24和TCCsup膀胱癌细胞株生长的影响,流式细胞仪检测其在诱导细胞凋亡与细胞周期捕获中的效果,APOPCYTO半胱天冬酶(caspase)色度法分析DAC处理后Caspase 3及Caspase 9的活性,蛋白质印迹法检测DAC处理后增殖细胞核抗原(PCNA)的表达情况.结果 DAC对4种膀胱肿瘤细胞的生长均具有抑制作用,药物作用呈剂量依存性.DAC不能诱导膀胱肿瘤细胞凋亡,细胞中Caspase 3、9活性未见激活,但DAC可以抑制膀胱癌细胞的增殖能力,PCNA蛋白表达量降低,发生G2/M期捕获.用0、1和8 μmol/L DAC处理膀胱癌细胞株RT112后G2/M期细胞分别为(36.3±3.4)%、(46.2±4.6)%和(56.5±6.2)%;处理TCCsup膀胱癌细胞株后为(37.5士3.8)%、(48.4±4.9)%和(60.1±6.7)%.结论 DAC对膀胱肿瘤细胞生长具有抑制作用,可能成为膀胱恶性肿瘤的一种新型治疗药物.
목적 탐토거갑기화약물5-담-2'-탈양포감(DAC)대방광종류세포생장적억제작용.방법 이용세포증식시제WST-1법측정불동농도DAC대RT112、253J、T24화TCCsup방광암세포주생장적영향,류식세포의검측기재유도세포조망여세포주기포획중적효과,APOPCYTO반광천동매(caspase)색도법분석DAC처리후Caspase 3급Caspase 9적활성,단백질인적법검측DAC처리후증식세포핵항원(PCNA)적표체정황.결과 DAC대4충방광종류세포적생장균구유억제작용,약물작용정제량의존성.DAC불능유도방광종류세포조망,세포중Caspase 3、9활성미견격활,단DAC가이억제방광암세포적증식능력,PCNA단백표체량강저,발생G2/M기포획.용0、1화8 μmol/L DAC처리방광암세포주RT112후G2/M기세포분별위(36.3±3.4)%、(46.2±4.6)%화(56.5±6.2)%;처리TCCsup방광암세포주후위(37.5사3.8)%、(48.4±4.9)%화(60.1±6.7)%.결론 DAC대방광종류세포생장구유억제작용,가능성위방광악성종류적일충신형치료약물.
Objective To study the growth suppressive effect of demethylation drug 5-aza-2'-deoxycytidine on bladder tumor cells. Methods The growth suppressive effect of DAC on 4 transitional cell carcinoma (TCC) cell lines was measured using the Cell Proliferation Reagent WST-1 assay.The effects of DAC on apoptosis induction and cell cycle arrest were analyzed by flow cytometric analysis. Caspase 3, 9 activities were analyzed by APOPCYTO Caspase Colorimetric Assay Kit and PCNA expression was also investigated by Western blot to clarify the mechanism of DAC against TCC. Results DAC inhibited the growth of all TCC cell lines tested in a dose-dependant manner, however,growth suppressive effect of DAC was independent of p53 status in TCC. DAC inhibited proliferation via inducing G2/M cell cycle arrest but not via inducing apoptosis. After treated with 0, 1 and 8 μmol/L DAC, cells of RTl 12 in G2/M phase was (36.3 ± 3.4) %, (46.2 ± 4.6) % and (56.5 ±6.2) %, TCCsup was (37.5 ± 3.8) %, (48.4 ±4.9) % and (60.1 ± 6.7) %, respectively. The expression of PCNA was decreased by DAC, but caspase3, 9 activities were not activated. Conclusion DAC could suppress the growth of TCC cells and might be a new strategy to treat bladder malignancy in the future.