长春新碱逆转骨髓间充质干细胞对白血病儿童门冬酰胺酶耐药的研究  

Vincristine Reverses the Resistance of Bone Marrow Mesenchy-mal Stem Cells to Asparaginase in Children with Leukemia

作　　者：徐清云[1] 卢婕伦 曾晓珍 卫风桂 吴泽霖[1] 邹亚伟[1] XU Qingyun;LU Jielun;ZENG Xiaozhen(Department of Pediatrics,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou City,Guangdong Province 510000)

机构地区：[1]广州医科大学附属第一医院儿科,广东省广州市510000

出　　处：《医学理论与实践》2022年第5期744-748,共5页The Journal of Medical Theory and Practice

基　　金：广东省自然科学基金项目(2016A030313654)。

摘　　要：目的:评估骨髓间充质干细胞(MSCs)对急性淋巴细胞白血病(ALL)患儿左旋门冬酰胺酶(L-asp)治疗的影响,并探讨长春新碱(VCR)能否逆转其影响及机制。方法:体外分离培养扩增ALL儿童MSCs原代细胞,建立MSCs-Jurkat共培养模型,应用MTT法检测Jurkat细胞L-asp的IC_(50)、流式细胞仪测定Jurkat细胞凋亡率及RT-PCR检测其ASNS mRNA表达。结果:(1)经L-asp处理后,单独培养及共培养组中Jurkat细胞凋亡率及ASNS mRNA表达水平均升高(P<0.05)。(2)共培养组中Jurkat细胞的凋亡率较Jurkat细胞单独培养低(P<0.01),ASNS mRNA表达水平降低(P<0.05);经L-asp处理后,共培养组中Jurkat细胞的凋亡率也比Jurkat单独培养低(P<0.01),但ASNS的mRNA表达水平无明显变化(P>0.05)。(3)VCR预处理MSCs后,共培养组中Jurkat细胞凋亡率升高(P<0.01),ASNS的mRNA表达水平无明显变化(P>0.05)。结论:MSCs与Jurkat细胞共培养可使Jurkat细胞凋率下降,抑制Jurkat细胞ASNS mRNA的表达。在MSCs支持下,L-asp对Jurkat细胞仍有作用,MSCs只可部分抑制L-asp细胞毒性。VCR可部分逆转MSCs的作用,使共培养模型中Jurkat细胞株凋亡增加,其机制可能是通过部分抑制MSCs ASNS mRNA表达。ABSTRACT Objective:To evaluate the impact of bone marrow mesenchymal stem cells(MSCs)on L-asp therapy in children with ALL,and to explore whether VCR can reverse its influence and mechanism.Methods:The primary cells of MSCs from ALL children were isolated,cultured and expanded in vitro,and MSCs-Jurkat co-culture model was established.The IC_(50) of L-asp of Jurkat cells was detected by MTT method,the apoptosis rate of Jurkat cells was measured by flow cytometry,and the ASNS mRNA expression was measured by RT-PCR.Results:(1)After treatment with L-asp,the apoptosis rate and ASNS mRNA expression of Jurkat cells in the single culture and co-culture groups increased(P<0.05).(2)The apoptosis rate of Jurkat cells in the co-culture group was lower than that of Jurkat cells cultured alone(P<0.01),and the ASNS mRNA expression level was reduced(P<0.05);after treatment with L-asp,the apoptosis of Jurkat cells in the co-culture group was also lower than Jurkat cells culture alone(P<0.01),but the mRNA expression level of ASNS did not change significantly(P>0.05).(3)After VCR pretreatment of MSCs,the apoptosis rate of Jurkat cells in the co-culture group increased(P<0.01),but the mRNA expression level of ASNS did not change significantly(P>0.05).Conclusion:Co-culture of MSCs and Jurkat cells can reduce the apoptotic rate of Jurkat cells and inhibit the expression of ASNS mRNA in Jurkat cells.With the support of MSCs,L-asp still has an effect on Jurkat cells,and MSCs can only partially inhibit L-asp cytotoxicity.VCR can partially reverse the effect of MSCs and increase the apoptosis of Jurkat cell line in the co-culture model,which may be related to the partial inhibition of mRNA expression of ASNS in MSCs.

关 键 词：骨髓间充质干细胞 门冬酰胺合成酶 白血病 左旋门冬酰胺酶 长春新碱 

分 类 号：R725.5[医药卫生—儿科]