中华外科杂志
中華外科雜誌
중화외과잡지
CHINESE JOURNAL OF SURGERY
2010年
5期
378-382
,共5页
居正华%应敏刚%艾星%史涛坪%王保军%王超%张国玺%张旭
居正華%應敏剛%艾星%史濤坪%王保軍%王超%張國璽%張旭
거정화%응민강%애성%사도평%왕보군%왕초%장국새%장욱
膀胱肿瘤%细胞系,肿瘤%5-氮杂脱氧胞苷%曲古抑菌素A%ALDH1a2基因
膀胱腫瘤%細胞繫,腫瘤%5-氮雜脫氧胞苷%麯古抑菌素A%ALDH1a2基因
방광종류%세포계,종류%5-담잡탈양포감%곡고억균소A%ALDH1a2기인
Urinary bladder neoplasms%Cell line,tumor%5-Aza-2'-deoxycitydine%Trichostatin A%ALDH1 a2 gene
目的 探讨5-氮杂脱氧胞苷(5-Aza-dC)与曲古抑菌素A(TSA)对膀胱癌细胞中抑癌基因ALDH1a2启动子甲基化状态和基因表达及细胞凋亡的影响.方法 使用5-Aza-dC、TSA处理RT-4、253J、5637、BIU-87和T24后,应用甲基化特异性PCR(MSP)法、逆转录PCR(RT-PCR)法、Western blot法分别检测这5株膀胱癌细胞经药物干预前后ALDH1a2甲基化状态及基因表达情况,应用流式细胞学检测干预前后5株膀胱癌细胞株的凋亡情况.结果 在5株膀胱癌细胞中,ALDH1a2基因表现为高甲基化表达受到抑制,TSA不影响甲基化状态,5-Aza-dC可逆转高甲基化状态并恢复基因表达,联合给予5-Aza-dC和TSA的作用和单独给予5-Aza-dC相似;TSA和5-Aza-dC均可诱导膀胱癌细胞株凋亡,早期凋亡是膀胱癌细胞株死亡的主要方式,5-Aza-dC和TSA有协同作用,3个实验组与对照组比较,差异均有统计学意义(P<0.05).结论 在5株膀胱癌细胞株中,ALDH1a2基因启动子甲基化可能是导致其基因失活的主要原因;5-Aza-dC单独作用和5-Aza-dC及TSA联合应用效果相似,均能恢复基因重新表达进而诱导膀胱癌细胞株的早期凋亡.
目的 探討5-氮雜脫氧胞苷(5-Aza-dC)與麯古抑菌素A(TSA)對膀胱癌細胞中抑癌基因ALDH1a2啟動子甲基化狀態和基因錶達及細胞凋亡的影響.方法 使用5-Aza-dC、TSA處理RT-4、253J、5637、BIU-87和T24後,應用甲基化特異性PCR(MSP)法、逆轉錄PCR(RT-PCR)法、Western blot法分彆檢測這5株膀胱癌細胞經藥物榦預前後ALDH1a2甲基化狀態及基因錶達情況,應用流式細胞學檢測榦預前後5株膀胱癌細胞株的凋亡情況.結果 在5株膀胱癌細胞中,ALDH1a2基因錶現為高甲基化錶達受到抑製,TSA不影響甲基化狀態,5-Aza-dC可逆轉高甲基化狀態併恢複基因錶達,聯閤給予5-Aza-dC和TSA的作用和單獨給予5-Aza-dC相似;TSA和5-Aza-dC均可誘導膀胱癌細胞株凋亡,早期凋亡是膀胱癌細胞株死亡的主要方式,5-Aza-dC和TSA有協同作用,3箇實驗組與對照組比較,差異均有統計學意義(P<0.05).結論 在5株膀胱癌細胞株中,ALDH1a2基因啟動子甲基化可能是導緻其基因失活的主要原因;5-Aza-dC單獨作用和5-Aza-dC及TSA聯閤應用效果相似,均能恢複基因重新錶達進而誘導膀胱癌細胞株的早期凋亡.
목적 탐토5-담잡탈양포감(5-Aza-dC)여곡고억균소A(TSA)대방광암세포중억암기인ALDH1a2계동자갑기화상태화기인표체급세포조망적영향.방법 사용5-Aza-dC、TSA처리RT-4、253J、5637、BIU-87화T24후,응용갑기화특이성PCR(MSP)법、역전록PCR(RT-PCR)법、Western blot법분별검측저5주방광암세포경약물간예전후ALDH1a2갑기화상태급기인표체정황,응용류식세포학검측간예전후5주방광암세포주적조망정황.결과 재5주방광암세포중,ALDH1a2기인표현위고갑기화표체수도억제,TSA불영향갑기화상태,5-Aza-dC가역전고갑기화상태병회복기인표체,연합급여5-Aza-dC화TSA적작용화단독급여5-Aza-dC상사;TSA화5-Aza-dC균가유도방광암세포주조망,조기조망시방광암세포주사망적주요방식,5-Aza-dC화TSA유협동작용,3개실험조여대조조비교,차이균유통계학의의(P<0.05).결론 재5주방광암세포주중,ALDH1a2기인계동자갑기화가능시도치기기인실활적주요원인;5-Aza-dC단독작용화5-Aza-dC급TSA연합응용효과상사,균능회복기인중신표체진이유도방광암세포주적조기조망.
Objective To study the effect on promoter de-methylation, expression of ALDH1a2 gene and cell apoptosis by treated with 5-Aza-dC and TSA in five human bladder cancer cell lines. Methods Human bladder cancer cell lines RT-4 ,253J, 5637, BIU-87 and T24 were cultured and treated with 5-AzadC and (or) TSA. The expression of the ALDH1a2 gene was detected by RT-PCR and Western blot. The methylation status of gene promoter was determined by MSP, and the cell cycle profile was established by flow cytometry. Results ALDH1a2 was silenced in five human bladder cancer cell lines. Re-expression of ALDH1a2 was detected after treated with 5-Aza-dC alone or TSA in combination. ALDH1a2 transcript was marked in each cell lines combined with 5-Aza-dC and TSA treatment which showed a synergistic effect on expression of ALDH1a2 transcript. Early apoptotic was the main mode of apoptosis and death of human bladder cancer cell lines induced by 5-Aza-dC and TSA. The percentage of early apoptotic cells was 1.4%in control group and 2. 8% in TSA group, however, 20. 2% in 5-Aza-dC group and 33.8% in 5-Aza-dC +TSA group, respectively. The groups of TSA, 5-Aza-dC and 5-Aza-dC + TSA were significantly different from control group (P < 0. 05 ). Conclusions Aberrant methylation of ALDH1 a2 gene is the main cause for gene transcriptional inactivation. Re-expression of ALDH1 a2 gene and cell apoptosis are detected after either treatment with 5-Aza-dC alone or in combination with TSA.
