LBPO.ET04 · 实验与分子治疗 · Late-Breaking
恢复端粒依赖性Hayflick极限以治疗血液系统恶性肿瘤
Restoring a telomere dependent Hayflick limit to treat hematological malignancies
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：尽管肿瘤细胞的端粒极短，但其持续增殖的后果是允许基因组不稳定性持续存在并传播至下一代细胞。基因组不稳定性是癌症的一个促成性标志。与正常细胞形成鲜明对比的是，肿瘤细胞通过激活端粒维持通路逃脱了谱系依赖性有限寿命的Hayflick极限。这些通路使细胞得以持续分裂，绕过Hayflick极限。癌细胞使用两种端粒维持通路来维持端粒长度。端粒酶激活是最常见的通路，为所有肿瘤细胞的85%所使用，而端粒替代延长（ALT）为其余部分所使用。一些肿瘤（尽管罕见）表现出两种通路的共激活。
方法：在我们的研究中使用了多种血液系统癌细胞系，包括：RPMI8226、MM.1R-CRL-2975（多发性骨髓瘤），MDS-L、MDS-92（骨髓增生异常综合征），OCI-AML3、Jurkat（急性白血病），以及人原代成纤维细胞（GL 51/92 Roe）。所用抑制剂为BIBR1532（72小时）以抑制端粒酶通路，Trabectedin（72-144小时）以抑制ALT通路。通过使用NucGreen™Dead488和台盼蓝排斥法的活细胞成像检测细胞死亡。
结果：在所有血液系统癌细胞系中，连续抑制两种端粒维持通路显示，经BIBR1532治疗72小时后细胞活力率显著下降。加入Trabectedin额外治疗3天后细胞死亡增加，在癌细胞中达到90%。相比之下，低传代和高传代的正常原代成纤维细胞（GL51/92 Roe）未表现出细胞死亡。
结论：在多种血液系统癌细胞系中使用小分子对两种端粒维持通路的序贯抑制显示细胞活力下降，而原代成纤维细胞的存活未受显著影响。未来使用原代癌细胞的研究有望证实这些结果，并为血液系统恶性肿瘤的治疗选择铺平道路。
关键词：肿瘤；基因组不稳定性；端粒酶；端粒替代延长（ALT）；细胞系；骨髓增生异常综合征（MDS）；急性白血病（AL）；多发性骨髓瘤；小分子
查看英文原文 English abstract
Background: The consequence of continued cell proliferation of tumor cells despite their critically short telomeres allows for ongoing genomic instability and its propagation to the next generation of cells. Genomic instability is an enabling hallmark of cancer . In stark contrast to normal cells, tumor cells escape the Hayflick limit of lineage-dependent limited life span through the activation of telomere maintenance pathways. These pathways allow cells to continue dividing, bypassing the Hayflick limit. Two telomere maintenance pathways are used by cancer cells to maintain telomere length. Telomerase activation is the most common pathway and is used by 85% of all tumor cells, while alternative lengthening of telomere (ALT) is used by the remainder. Some tumors, albeit rare, show the co-activation of both pathways.
Methods: We use in our study multiple hematological cancer cell lines including :RPMI8226, MM.1R-CRL-2975 (Multiple Myeloma), MDS-L, MDS-92 (Myelodysplastic syndromes), OCI-AML3, Jurkat (Acute Leukemia), and human primary fibroblasts (GL 51/92 Roe). The inhibitions used BIBR1532 (72 hours) to inhibit the telomerase pathway and Trabectedin(72-144hours) to inhibit the ALT pathway. Cell death was assayed by live cell imaging using NucGreen™Dead488 and trypan blue exclusion.
Results: Consecutive inhibition of both telomere maintenance pathways shows a significant decrease of cell viability rate over 72h of treatment with BIBR1532 in all hematological cancer cell lines. Cell death is increased by adding Trabectedin for an additional 3 days period and reaches 90 % in cancer cells. In contrast, the normal primary fibroblasts (GL51/92 Roe) at low and high passages do not show cell death.
Conclusion: The sequential inhibition of both telomere maintenance pathways using small molecules in multiples hematological cancer cell lines demonstrates a decrease in cell viability while primary fibroblasts show no significant effects on survival. Future studies using primary cancer cells are expected to confirm these results and to pave the way for cancer treatment options in hematological.
Keywords: neoplasia; genome instability; telomerase; alternative of lengthening telomere (ALT); cell lines; myelodysplastic syndromes (MDS); acute leukemia (AL); multiple myeloma; small molecules
利益披露 Disclosure
H. Sow, None..
S. Mai, None..
M. Gadji, None.