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Establishment of a bortezomib-resistant Chinese human multiple myeloma cell line: MMLAL

机译:耐硼替佐米的中国人多发性骨髓瘤细胞株MMLAL的建立

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Background 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. Methods 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. Results 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. Conclusion 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.
机译:背景从患有症状的国际III期IgG / lambda骨髓瘤的73岁中国患者的骨髓瘤性胸腔积液建立了新的人类骨髓瘤细胞系MMLAL。在短暂的完全缓解后,他发展为侵袭性全身复发,并伴有恶性胸腔积液和独家浆细胞浸润。他的疾病仍然是化学难治性的,并在复发六个月后死亡。方法在IL-6存在下培养患者胸腔积液中纯化的单核细胞。通过形态,免疫表型,细胞遗传,荧光原位杂交(FISH)和TP53突变分析来表征持续生长的细胞。在第3、6、9和12天测量细胞增殖并通过计数来与其他骨髓瘤细胞系进行比较。评估了对硼替佐米的耐药性,硼替佐米是一种批准用于符合条件的骨髓瘤患者的一线化疗的蛋白酶体抑制剂,并与其他骨髓瘤进行了比较MTT法检测细胞系。结果骨髓瘤细胞的免疫表型分析证实浆细胞标志物CD38和CD138的强表达,但T细胞或自然杀伤细胞标志物CD56的强表达。骨髓瘤细胞的细胞遗传学分析显示为二倍体复合核型,包括13号和17号染色​​体缺失或17号染色​​体的短臂缺失,即del(17p),呈等染色体17q10形式。 FISH证实具有TP53缺失但不存在t(4; 14)的二倍体核型。 TP53基因的测序分析表明没有突变。细胞计数表明最大活细胞密度约为2.5 X 10 6 细胞/ ml。硼替佐米治疗后,MTT分析显示IC 50 为72.17nM,表明强烈的硼替佐米耐药性。结论从中国一名骨髓瘤患者的胸腔积液中建立了一个二倍体,该染色体具有第13号染色体缺失和del(17p)缺失或人类骨髓瘤细胞株MMLAL。

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