Effects of Anti-CXCR4 Monoc lonal Antibody on Proliferation of Human Acute Myelocytic Leukemia Cell Line HL-60 Co-Cultured with Normal Bone Marrow Stromal Cells
-
摘要: 目的 :本文通过抗CXCR4单克隆抗体12G5阻断SDF-1活性,检测增殖相关指标,探讨SDF-1在维持HL-60细胞存活、增殖中的作用。 方法 :培养HL-60细胞并与正常骨髓基质细胞共培养,采用12G5(10μg/ml)阻断SDF-1生物学活性。孵育24h后,通过流式细胞仪观察HL-60细胞增殖周期的变化,免疫组化染色检测其PCNA蛋白表达的变化,并通过台盼蓝拒染观察白血病细胞存活、增殖的变化。 结果 :1)实验纽中G0/G1期细胞比例为(56.71±4.9)%,S期为(32.42±4.1)%,G2/M期为(10.87±5.2)%;对照组中G0/G1期细胞比例为(43.63±2.2)%,S期为(45.86±3.7)%,G2/M期为(10.51±2.6)%。2)台盼蓝拒染显示,12G5抑制HL-60细胞的存活及增殖。3)免疫组化染色结果显示,实验组及对照组中PCNA阳性细胞率分别为(42.1±3.9)%和(67.4±8.5)%。 结论 :SDF-1可在一定程度上维持HL-60细胞的存活及增殖进程。
-
关键词:
- SDF-1/CXCR4 /
- 单克隆抗体 /
- 基质细胞 /
- 骨髓 /
- 细胞增殖
Abstract: Objective :To study the effect of SDF-1 in proliferation of acute myeloeytic leukemia cells HL-60,the correlated index was analyzed when SDF-1 activity was blocked by anti-CXCR4 monoclonal antibody 12G5. Methods :HL-60 cells were cultured and co-cultured with normal stromal cells.24h after incubation with 12G5 (10μg/ml),cell cycle and PCNA expressions were detected with flow cytometry and immunohistochemical technique,respectivly.By means of rypan blue exclusion,cell survival rates were observed and growth curve was protracted. Results :The results were as follows:l) cell cycle in two groups were:experiment group G0/Gl (56.71±4.9)% ,S (32.42±4.1)%,G2/M (10.87±5.2)%, control group G0/Gl (43.63±2.2)%, S (45.86±3.7)%, G2/M (10.51±2.6)%, t-test analysis showed that 12GS induced more cells entering G0/Gl,phase.2) survival rates of HL-60 cells decreased markedly compared to control group,and cell proliferation slow down.3)immunohistoehemical technique shown: PCNA positive rates in experiment group and control group were (42.1土3.9)% and (67.4土8.5)% respectively,t-test analysis showed that 12GS down-regulated PCNA expressions. Conclusion :SDF-1 can keep survival and proliferation course of HL-60 cell at certain level.-
Keywords:
- SDF-1/CXCR4 /
- Monoclonal antibody /
- Stromal cell /
- Marrow /
- Cell Proliferation
-
-
[1] 1 Mudry RE, Fortney JE,York T, et la.Stromal cells regulate survival of B-linege leukemic cells during chemotherapy [J].Blood,2000,96(15):1926-1932
2 付相建,王苹,彭贤贵,等.急性白血病骨髓基质细胞对HL-60细胞增殖、分化的影响[J].重庆医学,2003,32(8):10l6-1018
3 Burger JA,Bonni A,West AE, et la. Chemokine receptors and stromal cells in the homing and homeostasis of chronic lymphoeycytic leukemia B cells [J].Leuk lymphoma,2002,43(3):461-466
4 Endres MJ,Clapham PR,Marsh M,et la.CD4-independent infection by HIV-2 is mediated by fusin/CXCR4 [J].Cell,1996,87(4):745-756
5 郭朝华,罗成基,孔佩艳,等.放烧复合伤小鼠骨髓基质细胞对粒单系祖细胞生长的影响[J].中国实验血液学杂志,1999,7(:)l65-68
6 Burger AJ,Tsukada N,Burger M,et al. Blood-derived-nurse like cells protect chronic lymphocytic leukemia B cells from spontaneous apoptosis through SDF-1[J],Blood 2000,96(8):2655-2663
7 Mohle R,Schittenhel M,Failenschmid C,et al.Functional response of leukemic blasts to SDF-1 correlates with preferential expression of the chemokine receptor CXCR4 in acute myelomonocytic and lymphoblasdtic leukemia [J].Br J Haematol,2000,110(3):563-572
8 Bradstock KF,Makrynikola V,Bianchi A, et al.Effects of the chemokine stromal cell-derived-factor-l on the migration localization of precursor-B acute lymphoblasdtic leukemia cells within bone marrow stromal layers[J].Leuk, 2000,14(5):882-888
9 Lee Y,Gotoh A,Kwon HJ,et la.Enhancement of intracellular signaling ssociated with hematopoietic progenitor cell survival in response to SDF-1/CXCL12 in synergy with other cytokines [J].Blood,2002,99,(12):4307-4317
10 Tsurusawa M,Ito M,Zha Z,et la. Cell-cycle-associated expression of proliferating cell nuclear antigen and Ki-67 reactive antigen of bone marrow blast cells in childhood acute leukemia [J].Luek,1992,6(7):669-674
计量
- 文章访问数: 2
- HTML全文浏览量: 0
- PDF下载量: 1
下载:
