Institute of Biomedicine, Department of Cellular Biology, Jinan University, No. 601 Huangpu Ave West, Shipai Street, Tianhe District, Guangzhou, Guangdong, 510632, China.
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China.
Biol Direct. 2024 Jun 24;19(1):49. doi: 10.1186/s13062-024-00490-1.
Most patients with acute myeloid leukemia (AML) eventually develop drug resistance, leading to a poor prognosis. Dysregulated long gene non coding RNAs (lincRNAs) have been implicated in chemoresistance in AML. Unfortunately, the effects of lincRNAs which participate in regulating the Adriamycin (ADR) resistance in AML cells remain unclear. Thus, the purpose of this study is to determine LINC00987 function in ADR-resistant AML.
In this study, ADR-resistant cells were constructed. LINC00987, miRNAs, and HMGA2 mRNA expression were measured by qRT-PCR. P-GP, BCRP, and HMGA2 protein were measured by Western blot. The proliferation was analyzed by MTS and calculated IC50. Soft agar colony formation assay and TUNEL staining were used to analyze cell colony formation and apoptosis. Xenograft tumor experiment was used to analyze the xenograft tumor growth of ADR-resistant AML.
We found that higher expression of LINC00987 was observed in AML patients and associated with poor overall survival in AML patients. LINC00987 expression was increased in ADR-resistant AML cells, including ADR/MOLM13 and ADR/HL-60 cells. LINC00987 downregulation reduces ADR resistance in ADR/MOLM13 and ADR/HL-60 cells in vitro and in vivo, while LINC00987 overexpression enhanced ADR resistance in MOLM13 and HL-60 cells. Additionally, LINC00987 functions as a competing endogenous RNA for miR-4458 to affect ADR resistance in ADR/MOLM13 and ADR/HL-60 cells. HMGA2 is a target of miR-4458. LINC00987 knockdown and miR-4458 overexpression reduced HMGA2 expression. HMGA2 overexpression enhanced ADR resistance, which reversed the function of LINC00987 silencing in suppressing ADR resistance of ADR/MOLM13 and ADR/HL-60 cells.
Downregulation of LINC00987 weakens ADR resistance by releasing miR-4458 to deplete HMGA2 in ADR/MOLM13 and ADR/HL-60. Therefore, LINC00987 may act as the therapeutic target for treating chemoresistant AML.
大多数急性髓系白血病（AML）患者最终会产生耐药性，导致预后不良。失调的长基因非编码 RNA（lncRNA）已被认为与 AML 中的化疗耐药有关。不幸的是，lncRNA 参与调节 AML 细胞中阿霉素（ADR）耐药性的作用仍不清楚。因此，本研究旨在确定 LINC00987 在 ADR 耐药 AML 中的功能。
在本研究中，构建了 ADR 耐药细胞。通过 qRT-PCR 测量 LINC00987、miRNAs 和 HMGA2 mRNA 的表达。通过 Western blot 测量 P-GP、BCRP 和 HMGA2 蛋白的表达。通过 MTS 分析增殖并计算 IC50。软琼脂集落形成试验和 TUNEL 染色用于分析细胞集落形成和细胞凋亡。异种移植肿瘤实验用于分析 ADR 耐药 AML 的异种移植肿瘤生长。
我们发现 AML 患者中 LINC00987 的表达较高，与 AML 患者的总生存期不良相关。ADR 耐药 AML 细胞中 LINC00987 的表达增加，包括 ADR/MOLM13 和 ADR/HL-60 细胞。LINC00987 下调降低了 ADR/MOLM13 和 ADR/HL-60 细胞在体外和体内的 ADR 耐药性，而 LINC00987 过表达增强了 MOLM13 和 HL-60 细胞的 ADR 耐药性。此外，LINC00987 作为 miR-4458 的竞争性内源 RNA 发挥作用，影响 ADR/MOLM13 和 ADR/HL-60 细胞的 ADR 耐药性。HMGA2 是 miR-4458 的靶标。LINC00987 下调和 miR-4458 过表达降低了 HMGA2 的表达。HMGA2 过表达增强了 ADR 耐药性，这逆转了 LINC00987 沉默在抑制 ADR/MOLM13 和 ADR/HL-60 细胞 ADR 耐药性中的作用。
下调 LINC00987 通过释放 miR-4458 耗尽 ADR/MOLM13 和 ADR/HL-60 中的 HMGA2，从而减弱 ADR 耐药性。因此，LINC00987 可能作为治疗化疗耐药性 AML 的治疗靶点。
