Department of Laboratory Medicine, Tokyo Medical and Dental University, Tokyo, Japan.
Department of Laboratory Medicine, Tokyo Medical and Dental University, Tokyo, Japan
Anticancer Res. 2022 Apr;42(4):1757-1761. doi: 10.21873/anticanres.15652.
BACKGROUND/AIM: TYRO3 is a member of the TAM family (TYRO3, AXL, and MERTK) of receptor tyrosine kinases. While the roles of activated AXL and MERTK in the growth of leukaemia cells have been reported, the effect of TYRO3 has not been determined. Therefore, we examined the effects of TYRO3 knockdown on the growth of leukaemia cell lines.
Three human leukaemia cell lines (AA derived from pure erythroid leukaemia, OCI/AML2, and K562), which express TYRO3 protein were used in this study. To induce TYRO3 knockdown, small interfering RNA (siRNA) against TYRO3 was transfected using an electroporation system. Cell growth was assessed by a colorimetric assay. The expression levels and activation of various signalling proteins were examined by immunoblotting. Changes in comprehensive gene expression after TYRO3 knockdown were examined by microarray analysis.
TYRO3 knockdown suppressed cell growth in the leukaemia cell lines tested. Additionally, the knockdown suppressed phosphorylation of signal transducer and activator of transcription-3 in AA cells, and extracellular signal-regulated kinase (ERK) 1/2 in AA and OCI/AML2 cells; both are downstream molecules of TYRO3 signalling. TYRO3 knockdown also suppressed the expression of survivin in all the cell lines. TYRO3 knockdown potently suppressed TYRO3 mRNA expression but not that of AXL and MERTK. Furthermore, TYRO3 knockdown suppressed cyclin D1 mRNA expression, which is a downstream molecule of ERK.
TYRO3 plays a role in leukaemia cell growth and is a potential therapeutic target for leukaemia.
背景/目的：TYRO3 是受体酪氨酸激酶 TAM 家族（TYRO3、AXL 和 MERTK）的成员。虽然已报道激活的 AXL 和 MERTK 在白血病细胞生长中的作用，但 TYRO3 的作用尚未确定。因此，我们研究了 TYRO3 敲低对白血病细胞系生长的影响。
本研究使用了三种表达 TYRO3 蛋白的人白血病细胞系（源自纯红细胞白血病的 AA、OCI/AML2 和 K562）。为了诱导 TYRO3 敲低，使用电穿孔系统转染针对 TYRO3 的小干扰 RNA（siRNA）。通过比色法评估细胞生长。通过免疫印迹检查各种信号蛋白的表达水平和激活情况。通过微阵列分析检查 TYRO3 敲低后综合基因表达的变化。
TYRO3 敲低抑制了所测试的白血病细胞系的细胞生长。此外，敲低抑制了 AA 细胞中转录激活因子 3 的磷酸化，以及 AA 和 OCI/AML2 细胞中细胞外信号调节激酶 1/2（ERK1/2）的磷酸化；两者都是 TYRO3 信号的下游分子。TYRO3 敲低还抑制了所有细胞系中生存素的表达。TYRO3 敲低强烈抑制 TYRO3 mRNA 表达，但不抑制 AXL 和 MERTK 的表达。此外，TYRO3 敲低还抑制了 ERK 的下游分子 cyclin D1 mRNA 的表达。
TYRO3 在白血病细胞生长中发挥作用，是白血病的潜在治疗靶点。
