人细胞周期蛋白C在儿童急性淋巴细胞性白血病中的表达及对白血病细胞增殖的影响

Expression of human cyclin C gene in childhood acute lymphocyte leukemia and its effect on leukemic cell proliferation

目的探讨人细胞周期蛋白C(human cyclin C,CCNC)在儿童急性淋巴细胞性白血病(acute lympho-cyte leukemia,ALL)中的表达,并初步研究CCNC在体外对白血病细胞增殖的影响。方法实时荧光定量PCR法检测CCNC在初治ALL、白血病细胞系及正常对照组中的表达,用脂质体Lifefectamine2000将真核表达载体PCMV-2B-CCNC转染至人急淋细胞系6T-CEM细胞株中,Western-blot检测转染后的细胞CCNC蛋白的表达量,流式细胞分析CCNC对白血病细胞增殖的作用。结果CCNC在对照组小儿、初治小儿ALL骨髓及6T-CEM细胞系中均有表达,在初治ALL中表达强度降低(P<0.05)。将PCMV-2B-CCNC转染至6T-CEM细胞中,Western-blot检测到转染后细胞CCNC蛋白表达量增高,流式细胞结果表明在转染后细胞中,Go/G1峰前出现一个亚二倍体峰,细胞凋亡指数有所增高(P<0.05)。结论CCNC在初治小儿ALL中出现较低的表达强度,在转染人白血病细胞后,具有诱导白血病细胞凋亡作用,有可能与儿童ALL的发病相关。

Objective To study the expression of human cyclin C （CCNC） gene in childhood acute lymphocyte leukemia （ALL） and to study its effect on leukemic cell proliferation in vitro. Methods Real -time -fluorescence quantitative PCR was used to quantitatively analyze the expression of CCNC. PCMV -2B/CCNC was subsequently transfected into the ALL 6T - CEM cell line using lipofactamine 2000. Western - blot was applied to detect the expression of CCNC of transfected cell and the changes of cell proliferation were observed by flow cytometry. Results CCNC could be detected in myeloid tissue of normal children and children with ALL and the expression was lower in children with ALL （ P 〈 0.05）. Western - blot proved CCNC had been transfected into the ALL 6T - CEM cell line. Flow cytometry showed that the apoptotic index of transfected cell increased （P 〈 0. 05）. Conclusion The expression of CCNC gene decreased in children with ALL and it might carry out a possible growth suppressive activity to leukemic cell in vitro, indicate its association with childhood ALL.