Department of Medicine, Queen Mary Hospital, University of Hong Kong, Room 419, Block K Pokfulam Road, Hong Kong, Hong Kong.
Cancer Cell Int. 2013 Dec 12;13(1):122. doi: 10.1186/1475-2867-13-122.
A new human myeloma cell line, MMLAL, was established from the myelomatous pleural effusion of a 73-year-old Chinese patient suffering from symptomatic International stage III IgG/lambda myeloma. After a brief period of complete remission, he developed aggressive systemic relapse complicated by malignant pleural effusion with exclusive plasma cell infiltration. His disease remained chemo-refractory, and died six months after relapse.
Purified mononuclear cells from the pleural effusion of the patient were cultured in the presence of IL-6. Continually growing cells were characterized by morphological, immunophenotypic, cytogenetic, fluorescence in situ hybridization (FISH) and TP53 mutation analyses. Cell proliferation was measured and compared with other myeloma cell lines by cell counting at day 3, 6, 9, and 12. Drug resistance against bortezomib, a proteasome inhibitor approved as a frontline chemotherapy for eligible myeloma patients, was evaluated and compared with other myeloma cell lines by MTT assay.
Immunophenotypic analysis of the myeloma cells confirmed strong expression of plasma cell markers CD38 and CD138 but not T-cell or natural killer-cell marker CD56. Cytogenetic analysis of the myeloma cells showed a hypodiploid composite karyotype including loss of chromosome 13 and 17 or deletion of the short arm of chromosome 17, i.e. del(17p), in the form of isochromosome 17q10. FISH confirmed a hypodiploid karyotype with TP53 deletion but absence of t(4;14). Sequencing analysis of the TP53 gene indicated absence of mutation. Cell counting revealed that the maximum viable cell density was about 2.5 X 106 cells/ml. Upon bortezomib treatment, MTT assay reported an IC50 of 72.17nM, suggesting a strong bortezomib resistance.
A hypodiploid with loss of chromosome 13 and loss or del(17p) human myeloma cell line, MMLAL, was established from the pleural effusion of a Chinese myeloma patient.
从一名 73 岁患有症状性国际分期 III 期 IgG/λ 骨髓瘤的中国患者的浆细胞瘤性胸腔积液中建立了一个新的人类骨髓瘤细胞系 MMLAL。在短暂的完全缓解后，他发生了侵袭性全身复发，伴有恶性胸腔积液，仅浆细胞浸润。他的疾病仍然对化疗有抗药性，复发后 6 个月死亡。
从患者的胸腔积液中纯化单核细胞，在 IL-6 的存在下培养。通过形态学、免疫表型、细胞遗传学、荧光原位杂交（FISH）和 TP53 突变分析来描述不断生长的细胞。通过细胞计数在第 3、6、9 和 12 天比较细胞增殖，并与其他骨髓瘤细胞系进行比较。通过 MTT 测定评估并比较硼替佐米（一种已被批准用于符合条件的骨髓瘤患者的一线化疗药物的蛋白酶体抑制剂）对骨髓瘤细胞的耐药性，与其他骨髓瘤细胞系进行比较。
骨髓瘤细胞的免疫表型分析证实了浆细胞标志物 CD38 和 CD138 的强烈表达，但不表达 T 细胞或自然杀伤细胞标志物 CD56。骨髓瘤细胞的细胞遗传学分析显示，低倍体复合核型包括 13 号染色体和 17 号染色体缺失或 17 号染色体短臂缺失，即 17q10 等臂染色体。FISH 证实低倍体核型存在 TP53 缺失但无 t(4;14)。TP53 基因测序分析表明无突变。细胞计数显示最大活细胞密度约为 2.5×106 个/ml。硼替佐米治疗后，MTT 测定报告 IC50 为 72.17nM，表明存在强烈的硼替佐米耐药性。
从一名中国骨髓瘤患者的胸腔积液中建立了一个低倍体、缺失 13 号染色体和缺失或 del(17p)的人类骨髓瘤细胞系 MMLAL。
