Analysis to Estimate Genetic Variations in the Idarubicin-Resistant Derivative MOLT-3.
Analysis to Estimate Genetic Variations in the Idarubicin-Resistant Derivative MOLT-3.
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DOI：
10.3390/ijms18010012
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发表时间：
2016-12-22
影响因子：
5.6
通讯作者：
Kobayashi H
中科院分区：
文献类型：
--
作者：
Komiyama T;Ogura A;Hirokawa T;Zhijing M;Kamiguchi H;Asai S;Miyachi H;Kobayashi H
基因改变是导致急性白血病细胞耐药性的一种公认机制。充分了解这些细胞的耐药机制将有助于更有效的化疗。在本研究中，我们通过完整的线粒体和核DNA分析，研究了人急性白血病细胞系MOLT-3及其伊达比星耐药衍生物MOLT-3/IDR的耐药机制。我们确定了这两种细胞系之间的遗传差异。ND 3突变位点（p.Thr61Ile）是MOLT-3/IDR细胞所特有的。此外，我们通过CGH阵列分析确定了5个具有遗传改变的候选基因，包括GALNT 2。GALNT 2外显子的测序显示，在MOLT-3/IDR细胞中终止密码子内存在G1716 K突变，但在MOLT-3细胞中不存在。这种突变导致GALNT 2编码的蛋白质中增加了18个氨基酸。使用实时PCR，我们确定该基因的表达值为0.35。蛋白质结构预测证实了MOLT-3/IDR细胞中GALNT 2的结构变化，对应于突变位点。我们推测该突变可能与艾达龙耐药有关。
Gene alterations are a well-established mechanism leading to drug resistance in acute leukemia cells. A full understanding of the mechanisms of drug resistance in these cells will facilitate more effective chemotherapy. In this study, we investigated the mechanism(s) of drug resistance in the human acute leukemia cell line MOLT-3 and its idarubicin-resistant derivative MOLT-3/IDR through complete mitochondrial and nuclear DNA analyses. We identified genetic differences between these two cell lines. The ND3 mutation site (p.Thr61Ile) in the mitochondrial DNA sequence was unique to MOLT-3/IDR cells. Moreover, we identified five candidate genes harboring genetic alterations, including GALNT2, via CGH array analysis. Sequencing of the GALNT2 exon revealed a G1716K mutation present within the stop codon in MOLT-3/IDR cells but absent from MOLT-3 cells. This mutation led to an additional 18 amino acids in the protein encoded by GALNT2. Using real-time PCR, we determined an expression value for this gene of 0.35. Protein structure predictions confirmed a structural change in GALNT2 in MOLT-3/IDR cells that corresponded to the site of the mutation. We speculate that this mutation may be related to idarubicin resistance.