[1]邓怡,杨世杰,贾延辉,等.Cx43修饰人脐血源基质细胞输注阻抑白血病MRD小鼠造血干细胞移植后复发[J].第三军医大学学报,2017,39(09):813-820.
 Deng Yi,Yang Shijie,Jia Yanhui,et al.Cx43 modified human umbilical cord blood-derived stromal cells prevent relapse in minimal residual disease model of mice after hematopoietic stem cells transplantation[J].J Third Mil Med Univ,2017,39(09):813-820.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第09期
页码:
813-820
栏目:
基础医学
出版日期:
2017-05-15

文章信息/Info

Title:
Cx43 modified human umbilical cord blood-derived stromal cells prevent relapse in minimal residual disease model of mice after hematopoietic stem cells transplantation
作者:
邓怡杨世杰贾延辉高蕾张诚刘耀司维柯张曦
第三军医大学新桥医院血液科,全军血液病中心;第三军医大学西南医院临床血液学教研室
Author(s):
Deng Yi Yang Shijie Jia Yanhui Gao Lei Zhang Cheng Liu Yao Si Weike Zhang Xi

Department of Hematology, Center of Hematology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037; Department of Clinical Hematology, Faculty of Laboratory Medicine, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China

关键词:
白血病复发微小残留病Cx43人脐血源基质细胞微环境
Keywords:
leukemia recurrence minimal residual disease connexin 43 human umbilical cord bloodderived stromal cells microenvironment
分类号:
R-332; R457; R733.7
文献标志码:
A
摘要:

目的     观察间隙连接蛋白43(connexin 43,Cx43)修饰人脐血源基质细胞(human umbilical cord blood-derived stromal cells,hUCBDSCs)体外对L615小鼠白血病细胞株凋亡以及在体对白血病微小残留病(minimal residual disease,MRD)小鼠疾病进展的影响。方法     通过Cx43过表达腺病毒(Ad-Cx43-GFP)上调hUCBDSCs中Cx43表达,体外构建L615+Cx43+hUCBDSCs共培养模型,检测其对L615细胞凋亡的影响。建立L615细胞低瘤负荷的MRD小鼠模型,分为骨髓(bone marrow,BM)移植组和Cx43+hUCBDSCs+BM移植组进行移植,以正常L615小鼠作为对照,检测移植后外周血象、骨髓涂片、组织病理及骨髓Cx43表达变化等。结果     Ad-Cx43-GFP能够在mRNA和蛋白水平显著上调hUCBDSCs中Cx43表达。L615+Cx43+hUCBDSCs移植组L615细胞凋亡比例较对照组显著升高[(8.93±1.24)% vs(3.53±0.13)%,P<0.01]。对MRD小鼠移植后,Cx43+hUCBDSCs+BM移植组外周血WBC和PLT恢复更快,17 d时接近正常水平,而BM移植组外周血WBC和PLT恢复延迟,17 d时低于正常水平;17 d时,Cx43+hUCBDSCs+BM移植组骨髓涂片原始细胞比例较BM移植组显著降低[(7.67±1.25)% vs (56.33±1.25)%,P<0.01];与BM移植组比较,Cx43+hUCBDSCs+BM移植组肝、脾、骨髓的白血病浸润程度较低,同时骨髓中Cx43蛋白表达增加。结论     上调hUCBDSCs中Cx43表达能在体外促进L615细胞凋亡,Cx43+hUCBDSCs+BM联合移植能够促进MRD小鼠外周血WBC和PLT恢复,阻抑MRD小鼠移植后复发。

Abstract:

Objective     To investigate the effect of connexin 43 (Cx43) modified human umbilical cord bloodderived stromal cells (hUCBDSCs) on the apoptosis of mouse leukemia cell line L615s and on pathoprogression in minimal residual disease (MRD) model of mice. Methods     Adenoviral plasmid Ad-Cx43-GFP was used to up-regulate the Cx43 expression in hUCBDSCs. The apoptotic rate of L615 cells co-cultured with Cx43-hUCBDSCs was determined by Annexin V/PI apoptosis detection kit. A mouse MRD model was generated through tail venous injection of with 1×105 GFP-L615 cells. Bone marrow (BM) and Cx43+hUCBDSCs+BM transplantation were performed in the MRD mice, and the model mice served as control. Peripheral blood cell count, BM pathological smears, histopathology and the expression of Cx43 in the BM were examined. Results     Ad-Cx43-GFP transfection significantly increased the expression of Cx43 in the hUCBDSCs at both mRNA and protein levels. The apoptotic rate of L615 cells was significantly higher in the Cx43+hUCBDSCs co-culture model than L615 cells cultured alone [(8.93±1.24)% vs (3.53±0.13)%, P<0.01]. Cx43+hUCBDSCs+BM transplantation resulted in obvious recoveries in the peripheral white blood cell count (WBC) and platelet count (PLT), with the counts almost reaching normal values in 17 d later. Whereas, the BM transplantation group showed delayed recoveries in peripheral WBC and PLT, and the counts were still lower than those of normal group in 17 d. The count of different bone marrow cells in BM pathological smears showed that the percentage of original cells was obviously decreased in the Cx43+hUCBDSCs+BM group than the BM group [(7.67±1.25)% vs (56.33±1.25)%, P<0.01]. The pathological observation showed an obviously lower infiltration of leukemia cells in the liver, spleen and BM of the Cx43+hUCBDSCs+BM group than that in the BM group. Furthermore, increased Cx43 in the BM was observed in the Cx43+hUCBDSCs+BM group than with the BM group. The expression of Cx43 in the bone marrow was enhanced in the Cx43+hUCBDSCs+BM group. Conclusion    Up-regulation of Cx43 in the hUCBDSCs can induce apoptosis in L615 cells. Combined Cx43 modified hUCBDSCs and hematopoietic stem cells transplantation can promote peripheral WBC and PLT recovery and prevent the relapse of MRD mice.

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更新日期/Last Update: 2017-05-04