| 摘要: | BT284是一甲醯胺衍生物，能夠抑制人類骨髓性血癌細胞HL-60之增生作用。本篇研究中，利用人類慢性骨髓性血癌細胞株K562能細胞分化與表現BCR-ABL 融合基因之特性，探討BT284影響K562分化的分子機制。先測試BT284對K562之細胞存活率，藥物之IC50約略於8-10 nM之間，故選用0-10 nM的濃度進行實驗。同時選用all-trans retinoic acid作為對照組藥物，利用May-Grunwald/Giemsa染色觀察細胞型態，以及使用四唑氮藍試驗檢測K562細胞在BT284處理過後是否進行細胞分化。在確認過BT284能夠促使K562細胞進行分化，觀察BT284同樣是否能影響血球分化相關基因之表現。選出紅血球、巨核細胞以及其他血球分化相關之基因: gata-1、nf-e2、gata-2、c-mpl 與c-myc，利用一步驟即時定量聚合?連鎖反應測定上述基因之mRNA表現。在BT284影響下，gata-1、gata-2、c-myc之mRNA表現增加，BT284同時也能抑制bcr-abl之mRNA表現。當BT284之濃度低於IC50時，能夠促使K562細胞進行分化。而癌細胞若進行分化，可能會伴隨著細胞凋亡。故此，未來BT284可成為抗血癌之藥物。
BT284 is a carboxamide analog which can inhibit the proliferation in human promyelocytic leukemia HL-60cell. To understand the molecular mechanisms of BT284 in human erythromyeloblastoid leukemia K562 cells, the differentiation in BT284 treated K562 cells are investigated. K562 cells were incubated with different concentrations of BT284 to explore the toxicity of BT284 by using MTT assay. After 24 h treatment of BT284, cell morphology changes were observed with May-Grunwald/Giemsa stain. Meanwhile, the effect of BT284 on differentiation was evaluated by real-time polymerase chain reaction and Nitro-blue tetrazolium (NBT) assay, respectively. IC50 of BT284 is around 10 nM in K562 for 24 h treatment. K562 cells underwent into differentiation with all trans retinoic acid and slightly morphology changes with different concentrations (0, 2, 4, 6, 8 and 10 nM) of BT284 by May-Grunwald/Giemsa stain. BT284 induced cell differentiation detected by NBT assay. Furthermore, expression levels of erythoid and megakaryocytic relative genes including gata-1, nf-e2, gata-2, c-mpl and c-myc were examined in BT-284 treated K562 cells. gata-1, gata-2 and c-myc were up-regulated by BT284. Since the K562 cell line is positive for the bcr-abl fusion gene, mRNA level of bcr-abl was assessed in BT-284 treated K562 cells. Surprisingly, data showed the inhibitory effect of BT284. In conclusion, cell morphology changed and differentiated under IC50 of BT284. Once differentiated, cells are triggering apoptosis. Afterwards, BT284 can be a candidate for treatment of cancer either in alone or in combination with other anti-cancer drugs. |
