School of Biomedicine, Faculty of Health Sciences, Universiti Sultan Zainal Abidin (UniSZA), Terengganu 21300, Malaysia.
Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan 16150, Malaysia.
Asian Pac J Cancer Prev. 2021 Dec 1;22(12):3959-3965. doi: 10.31557/APJCP.2021.22.12.3959.
BCR ABL oncogene encodes the BCR-ABL chimeric protein, which is a constitutively activated non-receptor tyrosine kinase. The BCR-ABL oncoprotein is a key molecular basis for the pathogenesis of chronic myeloid leukemia (CML) via activation of several downstream signaling pathways including JAK/STAT pathway. Development of leukemia involves constitutive activation of signaling molecules including, JAK2, STAT3, STAT5A and STAT5B. Thymoquinone (TQ) is a bioactive constituent of Nigella sativa that has shown anticancer properties in various cancers. The present study aimed to evaluate the effect of TQ on the expression of BCR ABL, JAK2, STAT3, STAT5A and STAT5B genes and their consequences on the cell proliferation and apoptosis in K562 CML cells.
BCR-ABL positive K562 CML cells were treated with TQ. Cytotoxicity was determined by Trypan blue exclusion assay. Apoptosis assay was performed by annexin V-FITC/PI staining assay and analyzed by flow cytometry. Transcription levels of BCR ABL, JAK2, STAT3, STAT5A and STAT5B genes were evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Protein levels of JAK2 and STAT5 were determined by Jess Assay analysis.
TQ markedly decreased the cell proliferation and induced apoptosis in K562 cells (P < 0.001) in a concentration dependent manner. TQ caused a significant decrease in the transcriptional levels of BCR ABL, JAK2, STAT3, STAT5A and STAT5B genes (P < 0.001). TQ induced a significant decrease in JAK2 and STAT5 protein levels (P < 0.001).
our results indicated that TQ inhibited cell growth of K562 cells via downregulation of BCR ABL/ JAK2/STAT3 and STAT5 signaling and reducing JAK2 and STAT5 protein levels.
BCR ABL 癌基因编码 BCR-ABL 嵌合蛋白，该蛋白是一种组成性激活的非受体酪氨酸激酶。BCR-ABL 癌蛋白是慢性髓系白血病（CML）发病机制的关键分子基础，通过激活包括 JAK/STAT 途径在内的几个下游信号通路。白血病的发展涉及包括 JAK2、STAT3、STAT5A 和 STAT5B 在内的信号分子的组成性激活。姜黄素（TQ）是黑种草中的一种生物活性成分，已在各种癌症中显示出抗癌特性。本研究旨在评估 TQ 对 BCR ABL、JAK2、STAT3、STAT5A 和 STAT5B 基因表达的影响及其对 K562 CML 细胞增殖和凋亡的影响。
用 TQ 处理 BCR-ABL 阳性 K562 CML 细胞。用台盼蓝排除试验测定细胞毒性。用 Annexin V-FITC/PI 染色法进行凋亡检测，并通过流式细胞术进行分析。通过逆转录定量聚合酶链反应（RT-qPCR）评估 BCR ABL、JAK2、STAT3、STAT5A 和 STAT5B 基因的转录水平。通过 Jess 分析测定 JAK2 和 STAT5 蛋白水平。
TQ 显著降低 K562 细胞的增殖并诱导细胞凋亡（P < 0.001），呈浓度依赖性。TQ 导致 BCR ABL、JAK2、STAT3、STAT5A 和 STAT5B 基因的转录水平显著降低（P < 0.001）。TQ 诱导 JAK2 和 STAT5 蛋白水平显著降低（P < 0.001）。
我们的结果表明，TQ 通过下调 BCR ABL/JAK2/STAT3 和 STAT5 信号通路和降低 JAK2 和 STAT5 蛋白水平来抑制 K562 细胞的生长。
