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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,h UCBDSCs)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h UCBDSCs中Cx43表达,体外构建L615+Cx43+h UCBDSCs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+h UCBDSCs+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h UCBDSCs中Cx43表达。L615+Cx43+h UCBDSCs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+h UCBDSCs+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+h UCBDSCs+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+h UCBDSCs+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h UCBDSCs中Cx43表达能在体外促进L615细胞凋亡,Cx43+h UCBDSCs+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。
Objective To observe the effects of connexin 43 (Cx43) on human umbilical cord blood-derived stromal cells (hUCBDSCs) in vitro on the apoptosis of leukemia cell line L615 and on the development of leukemia minimal residual disease (minimal residual disease, MRD) disease in mice. Methods The expression of Cx43 in UCBDSCs was induced by Cx43 overexpression adenovirus (Ad-Cx43-GFP), and the L615 + Cx43 + h UCBDSCs co-culture model was constructed in vitro. The MRD mouse model with low tumor burden of L615 cells was established and divided into bone marrow (BM) transplantation group and Cx43 + h UCBDSCs + BM transplantation group. Normal L615 mice were used as control to detect the peripheral blood, bone marrow Smears, histopathology and bone marrow Cx43 expression changes. Results Ad-Cx43-GFP significantly up-regulated Cx43 expression in UCBDSCs at mRNA and protein levels. The apoptosis ratio of L615 cells in L615 + Cx43 + h UCBDSCs transplantation group was significantly higher than that in control group [(8.93 ± 1.24)% vs (3.53 ± 0.13)%, P <0.01]. After transplanted into MRD mice, WBC and PLT recovered faster in peripheral blood of Cx43 + h UCBDSCs + BM transplantation group, approaching the normal level on the 17th day, while the recovery of WBC and PLT in the peripheral blood of BM transplantation group was delayed, and below the normal level on the 17th day ; On the 17th day, the ratio of bone marrow smear blasts in Cx43 + h UCBDSCs + BM group was significantly lower than that in BM group [(7.67 ± 1.25)% vs (56.33 ± 1.25)%, P <0.01] , Cx43 + h UCBDSCs + BM transplantation group liver, spleen, bone marrow leukemia infiltration is low, while bone marrow Cx43 protein expression increased. Conclusion Up-regulation of Cx43 expression in h UCBDSCs can promote the apoptosis of L615 cells in vitro. Cx43 + h UCBDSCs + BM transplantation can promote the recovery of WBC and PLT in peripheral blood of MRD mice and inhibit the recurrence of MRD mice after transplantation.