The inhibition of MARK2 suppresses cisplatin resistance of osteosarcoma stem cells by regulating DNA damage and repair
The inhibition of MARK2 suppresses cisplatin resistance of osteosarcoma stem cells by regulating DNA damage and repair
复制标题
抑制MARK2通过调节DNA损伤和修复来抑制骨肉瘤干细胞的顺铂耐药性
DOI：
10.1016/j.jbo.2020.100290
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发表时间：
2020-08-01
影响因子：
3.4
通讯作者：
Li, Jianmin
中科院分区：
文献类型：
--
作者：
Xu, Liang;Sun, Zhengkao;Li, Jianmin
目的：本研究旨在探讨MARK 2在CD 133（+）MG-63和MNNG/HOS细胞顺铂耐药模型中的作用及其机制。CCK-8法测定细胞活性。采用siRNA下调微管亲和力调节激酶2（MARK 2）表达。采用免疫荧光法和RT-qPCR法检测MARK 2和DNA-PKcs在蛋白和mRNA水平的表达。Western blot检测MARK 2、gamma H2 AX（S139）、DNA-PKcs、Phospho-PI 3 Kinase p85（Tyr 458）、Akt、phospho-Akt（T308）抗体、mTOR、phospho-mTOR（Ser 2448）抗体水平。与CD 133- MG-63细胞相比，CD 133（+）MG-63细胞对顺铂的耐药性明显增强，MARK 2表达水平升高，DNA-PKcs和强有力的DNA损伤修复能力（p
Objective: This study aims to explore the role of MARK2 in chemotherapeutic resistance and potential mechanism within cisplatin resistance models of CD133(+) MG-63 and MNNG/HOS cells.Methods: CD133(-) and CD133(+) MG-63 and MNNG/HOS cells were differentiated and obtained by MACS (Magnetic bead sorting). Cell activity was determined by CCK-8 assay. siRNA was employed to down regulate the Microtubule Affinity Regulated Kinase 2 (MARK2) expression. Immunofluorescence detection and RT-qPCR were used to measure the expressions of MARK2 and DNA-PKcs at both protein and mRNA levels. Western blot was applied to test the levels of MARK2, gamma H2AX (S139), DNA-PKcs, Phospho-PI3 Kinase p85 (Tyr458), Akt, phosphoAkt (T308) antibodies, mTOR, phospho-mTOR (Ser2448).Results: Compared with CD133- MG-63 cells, CD133(+) MG-63 cells showed significantly strong cisplatin resistance, with high levels of MARK2, DNA-PKcs and potent DNA damage repair ability (p