前B急性淋巴细胞白血病细胞Sup-B15对化疗药物敏感性及侵袭力的研究

Chemosensitivity and angiogenic potency of pre-B acute lymphoblastic leukemia cell line Sup-B15

目的 探讨Ik6阳性表达的前B急性淋巴细胞白血病细胞Sup-B15对化疗药物的敏感性及侵袭力,并与国内常用的细胞模型Nalm-6进行比较研究.方法 采用巢式反转录-PCR及Western blot法检测细胞Ik6的表达;应用CCK-8法检测化疗药物长春新碱（VCR）、柔红霉素（DNR）、左旋门冬酰胺酶（L-Asp）及地塞米松（Dex）对细胞增殖的影响;采用Transwell小室检测细胞的迁移和侵袭力;应用实时荧光定量PCR法检测血管新生相关基因VEGF、Flt-1、PlGF、Ang1、Ang2和IGF-1的表达.结果 Nalm-6细胞中未检测到Ik6的表达.相比较于Nalm-6细胞,Sup-B15细胞生长速度较慢.4种化疗药物均能以时间和剂量依赖方式抑制两细胞的增殖,Sup-B15细胞对DNR、L-Asp和Dex更敏感,IC50分别是Nalm-6细胞的9.5倍、1.1倍和1.5倍,但是对VCR的敏感性不及Nalm-6细胞.Transwell小室实验显示迁移或侵袭到下室的Sup-B15细胞较少.血管新生相关基因在Sup-B15细胞中相对低表达.结论 Sup-B15细胞相比较于Nalm-6细胞,是一种对化疗药物更敏感、侵袭力较弱的前B急性淋巴细胞系.

Objective It has been reported that Ik6 overexpression is associated with poor outcome of childhood pre-B acute lymphoblastic leukemia and Sup-B15 cell line is Ik6 positive.There were fewer studies on the Sup-B15 in the country,so the aim of this research is to explore chemosensitivity and angiogenic potency of pre-B acute lymphoblastic leukemia cell line Sup-B15 associated with Ik6 positive expression through making a comparison between Sup-B15 and Nalm-6 cell line.Methods The Ik6 mRNA and protein expression were detected by nested reverse transcription polymearse chain reaction and Western blot.The effect of vincristine,daunorubicin,L-asparaginase and dexamethasone on the proliferation of cells was measured by using CCK-8 assay.The migration and invasion of cells were detected by transwell assay and mRNA expression of angiogenesis genes （VEGF,Flt-1,PlGF,IGF-1,Ang1 and Ang2） was measured by real-time PCR.Results No expression of Ik6 was detected in Nalm-6 cell line.Compared with Nalm-6,Sup-B15 cell line had a lower rate of growth.The 4 drugs could inhibit the proliferation of cells in a dose-and time-depen-dence manner.Sup-B15 was more sensitive to daunorubicin,L-asparaginase and dexamethasone,and the folds of respective IC50 value were 9.5,1.1 and 1.5.But Sup-B15 was more resistant to vincristine than Nalm-6.The result of transwell assay showed that less Sup-B15 cells transmigrated to the lower chamber.A significant lower expression level of angiogenesis genes was detected in Sup-B15 cells.Conclusion These findings suggest that Sup-B15 is more sensitive to chemotherapy and less angiogenic compared with Nalm-6.