中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
1期
17-21
,共5页
韩旭%谭志军%郭仁德%李照晋%王玉亮
韓旭%譚誌軍%郭仁德%李照晉%王玉亮
한욱%담지군%곽인덕%리조진%왕옥량
5-氮杂-2’-脱氧胞苷%胰腺肿瘤%RUNX3基因%甲基化
5-氮雜-2’-脫氧胞苷%胰腺腫瘤%RUNX3基因%甲基化
5-담잡-2’-탈양포감%이선종류%RUNX3기인%갑기화
5-aza-2'-deoxycytidine%Pancreatic neoplasms%Gene,RUNX3%Methylation
目的 探讨去甲基化制剂5-氮杂-2’-脱氧胞苷(5-Aza-CdR)对胰腺癌细胞系MiaPaca2生长的影响,以及对RUNX3抑癌基因表达和甲基化的影响.方法 光镜观察5-Aza-CdR处理前后MiaPaca2细胞形态学变化,四甲基偶氮唑蓝法检测5-Aza-CdR对MiaPaca2细胞增殖活性的影响,半定量逆转录聚合酶链反应检测RUNX3 mRNA表达的变化,甲基化特异性聚合酶链反应检测RUNX3基因甲基化的变化.结果 经2.5、5、10和20 μmol/L 5-Aza-CdR处理24h后,MiaPaca2细胞的抑制率分别为(9.17 ±2.15)％、(10.75 ±2.04)％、(12.57±1.64)％和(18.70±1.51)％;48 h后分别为(14.94±1.68)％、(18.60±1.57)％、(22.84±1.58)％和(33.24±1.53)％；72 h后分别为(21.46±1.60)％、(28.62±1.72)％、(35.14±1.64)％和(45.06±1.47)％；96 h后分别为(26.35±1.71)％、(34.48±1.69)％、(40.05 ±1.60)％和(49.99±1.61)％.各实验组与对照组抑制率比较,差异均有统计学意义(均P ＜0.05).在同一时点,各浓度组间MiaPaca2细胞的抑制率差异均有统计学意义(均P＜0.05).在48、72、96h时,各浓度组抑制率两两比较,差异均有统计学意义(均P＜0.05).在药物处理24h以后,5-Aza-CdR对MiaPaca2细胞的生长抑制作用随着5-Aza-CdR浓度的增加而增强.5-Aza-CdR能够逆转RUNX3基因的甲基化状态,恢复RUNX3 mRNA的表达.结论 RUNX3基因甲基化状态与胰腺癌的发生发展密切相关,异常甲基化可能是引起RUNX3基因表达缺失的重要机制.5-Aza-CdR可以逆转MiaPaca2细胞RUNX3基因的甲基化状态,重新激活其表达,从而抑制肿瘤细胞的生长,为胰腺癌的诊断和治疗提供了一条新途径.
目的 探討去甲基化製劑5-氮雜-2’-脫氧胞苷(5-Aza-CdR)對胰腺癌細胞繫MiaPaca2生長的影響,以及對RUNX3抑癌基因錶達和甲基化的影響.方法 光鏡觀察5-Aza-CdR處理前後MiaPaca2細胞形態學變化,四甲基偶氮唑藍法檢測5-Aza-CdR對MiaPaca2細胞增殖活性的影響,半定量逆轉錄聚閤酶鏈反應檢測RUNX3 mRNA錶達的變化,甲基化特異性聚閤酶鏈反應檢測RUNX3基因甲基化的變化.結果 經2.5、5、10和20 μmol/L 5-Aza-CdR處理24h後,MiaPaca2細胞的抑製率分彆為(9.17 ±2.15)％、(10.75 ±2.04)％、(12.57±1.64)％和(18.70±1.51)％;48 h後分彆為(14.94±1.68)％、(18.60±1.57)％、(22.84±1.58)％和(33.24±1.53)％；72 h後分彆為(21.46±1.60)％、(28.62±1.72)％、(35.14±1.64)％和(45.06±1.47)％；96 h後分彆為(26.35±1.71)％、(34.48±1.69)％、(40.05 ±1.60)％和(49.99±1.61)％.各實驗組與對照組抑製率比較,差異均有統計學意義(均P ＜0.05).在同一時點,各濃度組間MiaPaca2細胞的抑製率差異均有統計學意義(均P＜0.05).在48、72、96h時,各濃度組抑製率兩兩比較,差異均有統計學意義(均P＜0.05).在藥物處理24h以後,5-Aza-CdR對MiaPaca2細胞的生長抑製作用隨著5-Aza-CdR濃度的增加而增彊.5-Aza-CdR能夠逆轉RUNX3基因的甲基化狀態,恢複RUNX3 mRNA的錶達.結論 RUNX3基因甲基化狀態與胰腺癌的髮生髮展密切相關,異常甲基化可能是引起RUNX3基因錶達缺失的重要機製.5-Aza-CdR可以逆轉MiaPaca2細胞RUNX3基因的甲基化狀態,重新激活其錶達,從而抑製腫瘤細胞的生長,為胰腺癌的診斷和治療提供瞭一條新途徑.
목적 탐토거갑기화제제5-담잡-2’-탈양포감(5-Aza-CdR)대이선암세포계MiaPaca2생장적영향,이급대RUNX3억암기인표체화갑기화적영향.방법 광경관찰5-Aza-CdR처리전후MiaPaca2세포형태학변화,사갑기우담서람법검측5-Aza-CdR대MiaPaca2세포증식활성적영향,반정량역전록취합매련반응검측RUNX3 mRNA표체적변화,갑기화특이성취합매련반응검측RUNX3기인갑기화적변화.결과 경2.5、5、10화20 μmol/L 5-Aza-CdR처리24h후,MiaPaca2세포적억제솔분별위(9.17 ±2.15)％、(10.75 ±2.04)％、(12.57±1.64)％화(18.70±1.51)％;48 h후분별위(14.94±1.68)％、(18.60±1.57)％、(22.84±1.58)％화(33.24±1.53)％；72 h후분별위(21.46±1.60)％、(28.62±1.72)％、(35.14±1.64)％화(45.06±1.47)％；96 h후분별위(26.35±1.71)％、(34.48±1.69)％、(40.05 ±1.60)％화(49.99±1.61)％.각실험조여대조조억제솔비교,차이균유통계학의의(균P ＜0.05).재동일시점,각농도조간MiaPaca2세포적억제솔차이균유통계학의의(균P＜0.05).재48、72、96h시,각농도조억제솔량량비교,차이균유통계학의의(균P＜0.05).재약물처리24h이후,5-Aza-CdR대MiaPaca2세포적생장억제작용수착5-Aza-CdR농도적증가이증강.5-Aza-CdR능구역전RUNX3기인적갑기화상태,회복RUNX3 mRNA적표체.결론 RUNX3기인갑기화상태여이선암적발생발전밀절상관,이상갑기화가능시인기RUNX3기인표체결실적중요궤제.5-Aza-CdR가이역전MiaPaca2세포RUNX3기인적갑기화상태,중신격활기표체,종이억제종류세포적생장,위이선암적진단화치료제공료일조신도경.
Objective To investigate the effect of demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human pancreatic cancer cell line MiaPaca2 and the expression and methylation of tumor suppressor gene RUNX3.Methods Human pancreatic cancer cell line MiaPaca2 cells were treated with different concentrations of 5-Aza-CdR.Morphological changes of MiaPaca2 cells were observed by light microscopy.The activity of cell proliferation was analyzed by MTT assay.The changes of RUNX3 mRNA expression were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).Changes of RUNX3 gene methylation was detected by methylation-specific polymerase chain reaction.Results MiaPaca2 cells were treated with 2.5,5,10 and 20 μ mol/L 5-Aza-CdR,respectively.The inhibition rates of MiaPaca2 cells treated for 24 h were (9.17 ±2.15)％,(10.75 ±2.04)％,(12.57 ±1.64)％ and (18.70±1.51)％,respectively.The inhibition rates were (14.94 ±1.68)％,(18.60 ±1.57)％,(22.84 ± 1.58)％ and (33.24 ± 1.53)％,respectively,after 48 h treatment; (21.46 ±1.60)％,(28.62 ± 1.72)％,(35.14 ± 1.64)％ and (45.06 ± 1.47)％,respectively,after 72 htreatment; and (26.35 ±1.71)％,(34.48 ± 1.69)％,(40.05 ± 1.60)％ and (49.99 ± 1.61)％,respectively,after 96 h treatment.The differences between inhibition rates of each experimental and control groups (0.00 ± 0.00) ％ were statistically significant (P ＜ 0.05).At the same time,the inhibition rates of different concentration groups showed significant differences (P ＜ 0.05).At 48 h,72 h and 96 h,the inhibition rates of each pair concentration groups showed significant differences (P ＜ 0.05).5-Aza-CdR inhibited the growth of MiaPaca2 cells,and the higher the concentration,the stronger the inhibition after 24 h.5-Aza-CdR also reversed the methylation status of RUNX3 gene,and restored the expression of RUNX3 mRNA with a dose-effect relationship.Conclusions The methylation of RUNX3 gene is significantly related with the occurrence and development of pancreatic cancer,and abnormal methylation of RUNX3 gene may contribute to the loss of RUNX3 mRNA expression.5-Aza-CdR may effectively cause reversion of RUNX3 methylation,and treatment with 5-Aza-CdR can reactivate the gene expression and inhibit the cell growth.This may provide a new way for diagnosis and treatment of pancreatic cancer.
