白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2013年
11期
678-681
,共4页
林聪猛%梅序桥%郑源海%林福安%叶宝国
林聰猛%梅序橋%鄭源海%林福安%葉寶國
림총맹%매서교%정원해%림복안%협보국
白血病,淋巴细胞,急性%丙戊酸钠%原代细胞%p15基因%甲基化
白血病,淋巴細胞,急性%丙戊痠鈉%原代細胞%p15基因%甲基化
백혈병,림파세포,급성%병무산납%원대세포%p15기인%갑기화
Leukemia,lymphocytic,acute%Valproate%Primary cells%p15 gene%Methylation
目的 探讨丙戊酸钠(VPA)对急性淋巴细胞白血病(ALL)原代细胞凋亡现象与去甲基化机制之间的联系.方法 选取10例AIL患者原代细胞,MTT法检测VPA对ALL原代细胞增殖抑制作用,DNA ladder试验观察细胞凋亡,Annexin-V-FITC/PI检测细胞早期凋亡,半巢式甲基化特异PCR检测p15INK4B基因甲基化,反转录PCR方法检测p15基因表达水平.实验数据采用SPSS 16.0统计软件处理.结果 VPA可明显抑制细胞增殖,对原代细胞IC50为1.898 mmol/L;DNA ladder试验显示VPA对原代ALL细胞的凋亡作用明显.Annexin-V-FITC/PI检测1.0、2.0、4.0 mmol/LVPA组的凋亡率分别是(5.80±0.65)％、(48.46±2.49)％、(76.45±2.98)％,与未用药组的(0.44±0.04)％比较差异有统计学意义(P＜0.05).原代ALL细胞经VPA作用后,p15INK4B甲基化水平明显下降,2.0、4.0 mmol/LVPA组与未用药组相比,p15 mRNA表达水平增强,差异有统计学意义(P＜0.05).结论 VPA通过使p15INK4B基因去甲基化,促进p15基因表达上调,从而促进原代ALL细胞凋亡.
目的 探討丙戊痠鈉(VPA)對急性淋巴細胞白血病(ALL)原代細胞凋亡現象與去甲基化機製之間的聯繫.方法 選取10例AIL患者原代細胞,MTT法檢測VPA對ALL原代細胞增殖抑製作用,DNA ladder試驗觀察細胞凋亡,Annexin-V-FITC/PI檢測細胞早期凋亡,半巢式甲基化特異PCR檢測p15INK4B基因甲基化,反轉錄PCR方法檢測p15基因錶達水平.實驗數據採用SPSS 16.0統計軟件處理.結果 VPA可明顯抑製細胞增殖,對原代細胞IC50為1.898 mmol/L;DNA ladder試驗顯示VPA對原代ALL細胞的凋亡作用明顯.Annexin-V-FITC/PI檢測1.0、2.0、4.0 mmol/LVPA組的凋亡率分彆是(5.80±0.65)％、(48.46±2.49)％、(76.45±2.98)％,與未用藥組的(0.44±0.04)％比較差異有統計學意義(P＜0.05).原代ALL細胞經VPA作用後,p15INK4B甲基化水平明顯下降,2.0、4.0 mmol/LVPA組與未用藥組相比,p15 mRNA錶達水平增彊,差異有統計學意義(P＜0.05).結論 VPA通過使p15INK4B基因去甲基化,促進p15基因錶達上調,從而促進原代ALL細胞凋亡.
목적 탐토병무산납(VPA)대급성림파세포백혈병(ALL)원대세포조망현상여거갑기화궤제지간적련계.방법 선취10례AIL환자원대세포,MTT법검측VPA대ALL원대세포증식억제작용,DNA ladder시험관찰세포조망,Annexin-V-FITC/PI검측세포조기조망,반소식갑기화특이PCR검측p15INK4B기인갑기화,반전록PCR방법검측p15기인표체수평.실험수거채용SPSS 16.0통계연건처리.결과 VPA가명현억제세포증식,대원대세포IC50위1.898 mmol/L;DNA ladder시험현시VPA대원대ALL세포적조망작용명현.Annexin-V-FITC/PI검측1.0、2.0、4.0 mmol/LVPA조적조망솔분별시(5.80±0.65)％、(48.46±2.49)％、(76.45±2.98)％,여미용약조적(0.44±0.04)％비교차이유통계학의의(P＜0.05).원대ALL세포경VPA작용후,p15INK4B갑기화수평명현하강,2.0、4.0 mmol/LVPA조여미용약조상비,p15 mRNA표체수평증강,차이유통계학의의(P＜0.05).결론 VPA통과사p15INK4B기인거갑기화,촉진p15기인표체상조,종이촉진원대ALL세포조망.
Objective To investigate the relation between demethylation effect and apoptosis of valproate (VPA) in primary acute lymphoblastic leukemia (ALL) cells in vitro.Methods 10 cases of ALL patients was choosed to acquire leukemia cells.Cell growth curve were assessed by the MTT assay,the apoptosist of primary ALL was analyzed with DNA Ladder and Annexin-V-FITC/PI by flow cytometry.The expression methylation level of p15 was detected by hn-MSPCR,and p15mRNA were detected by RT-PCR.All dates was analyzed by SPSS16.0.Results The 50 ％ inhibition rate of VPA were 1.898 mmol/L to primary ALL cells assayed by MTT respectively.DNA ladder showed the apoptosis of primary ALL cells increased by adding VPA dose.Annexin-V-FITC/PI tests showed that the apoptosis percentage of primary ALL cells were (0.44±0.04) ％ in control group,(5.80±0.65) ％ in 1.0 mmol/L VPA group,(48.46±2.49) ％ in 2.0 mmol/L VPA group,(76.45±2.98) ％ in 4.0 mmol/L VPA group,the apoptosis percentage increased significantly (P ＜ 0.05).The demethylation of p15 INK4B gene decreased by adding VPA dose,the expression of p15 mRNA expression increased significantly compared with control group by RT-PCR (P ＜ 0.05).Conclusion It is found that VPA could induce demethylation of p15 INK4B gene,which could upregulate the p15 mRNA expression,due to the apoptosis of primary ALL cells.
