Isolation and characterization of a CD34+ sub-clone in B-cell lymphoma.
Isolation and characterization of a CD34+ sub-clone in B-cell lymphoma.
复制标题
DOI：
10.18632/oncotarget.27415
复制
发表时间：
2020-01-14
期刊：
影响因子：
--
通讯作者：
Gabali, Ali M
中科院分区：
文献类型：
--
作者：
Al-Katib, Ayad M;Ebrahim, Abdul Shukkur;Gabali, Ali M
非霍奇金淋巴瘤（NHL）是美国最常见的血液恶性肿瘤。尽管对初始治疗有反应并达到完全缓解（CR），但许多类型仍然无法治愈。先进的实验室技术，如多色流式细胞术（FCM）和聚合酶链反应（PCR）已经证明了治疗后罕见的恶性细胞群的持久性。然而，该种群的功能和生物学特性尚未阐明。建立的b -淋巴瘤细胞系（B-NHL）和患者来源的样本（PDS）采用8色流式细胞仪进行分析。CD34+亚群通过体外暴露于2-氯脱氧腺苷（2-CdA）和CD34磁珠进行富集。通过核型分析和阵列比较基因组杂交（aCGH）对细胞组分进行遗传分析。通过短期体外化疗暴露来测定对化疗的敏感性。采用软琼脂集落形成法测定克隆原性，碘化丙啶DNA染色法和流式细胞仪测定增殖能力。流式细胞术显示，在B-NHL细胞系（3 / 3）和PDS细胞系（8 / 8）中，存在表达CD34的单型b细胞的微小亚克隆。通过暴露于2-CdA，该亚群在体外富集高达50倍，通过CD34磁珠柱分离纯度高达80%。除CD34表达外，该群体表达与亲本细胞相同的表型和基因型，但与CD34-群体相比，其增殖性更强，Hoechst 33342阳性，克隆性更强，耐化疗。分离的CD34+单型b细胞可能有助于某些NHL对治疗的耐药，应该成为潜在的NHL新药的靶点。
Non-Hodgkin's lymphoma (NHL) is the most common hematological malignancy in the US. Many types remain incurable despite response to initial therapy and achievement of complete remission (CR). Advanced laboratory techniques like multicolor flow cytometry (FCM) and polymerase chain reaction (PCR) have demonstrated persistence of rare malignant cell population post therapy. However, the functional and biological characteristics of this population have not been elucidated. Established B-lymphoma cell lines (B-NHL) and patient-derived samples (PDS) were analyzed using 8-color FCM. CD34+ sub-population was enriched using in vitro exposure to 2-chlorodeoxyadenosine (2-CdA) and by CD34 magnetic beads. Genetic analysis of cell fractions was done by karyotyping and array comparative genomic hybridization (aCGH). Sensitivity to chemotherapy was assayed by short-term in vitro exposure to chemotherapy. Clonogenicity was determined by soft agar colony formation assay, and proliferation was determined using DNA staining with propidium iodide and FCM. FCM demonstrated the presence of a minute sub-clone of monotypic B-cells that express CD34 in B-NHL cell lines (3 of 3) and in PDS (8 of 8). This sub-population enriched up to 50 fold in vitro by exposure to 2-CdA and up to 80% purity by CD34 magnetic bead column isolation. Except for CD34 expression, this population expressed identical phenotype and genotype to parent cells, but was more proliferative, Hoechst 33342-positive, clonogenic, and resistant to chemotherapy compared with the CD34- population. The isolated CD34+ monotypic B-cells may contribute to resistance of certain NHL to treatment and should be targeted by potential new drugs for NHL.