Chen Li, Zhao Hongmian, Wang Chao, Hu Ning
Department of Hematology, Huaihe Hospital of Henan University No. 115 Ximen Street Kaifeng 475000 Henan China
RSC Adv. 2019 Apr 8;9(19):10897-10904. doi: 10.1039/c9ra00306a. eCollection 2019 Apr 3.
Taurine-upregulated gene 1 (TUG1) has been reported as an oncogenic long non-coding RNA (lncRNA) in acute myeloid leukemia (AML). Nevertheless, the roles and molecular mechanism of TUG1 in drug resistance of AML cells are still unclear. Glycolysis level was evaluated by detecting glucose consumption and lactate production. qRT-PCR and Western blot were performed to detect TUG1, hexokinase2 (HK2) and pyruvate kinase isoenzyme M2 (PKM2) expressions. Adriamycin (ADR) cytotoxicity and apoptosis were assessed by MTT assay and flow cytometry, respectively. The changes of the protein kinase B (Akt) pathway were determined by Western blot analysis of phosphorylated-Akt (p-Akt) (ser473) and Akt. Our results showed that glycolysis was increased in HL60/ADR cells, as evidenced by the elevated glucose consumption and lactate production, as well as the increased HK2 and PKM2 expressions at mRNA and protein levels. TUG1 was up-regulated in HL60/ADR cells and TUG knockdown inhibited glycolysis. TUG1 knockdown enhanced ADR-induced cytotoxicity and apoptosis in HL60/ADR cells. TUG1 knockdown inhibited the Akt pathway and activation of the Akt pathway by 740Y-P attenuated the effects of TUG1 knockdown on ADR-induced cytotoxicity and apoptosis, as well as glycolysis in HL60/ADR cells. Taken together, TUG1 knockdown enhances adriamycin cytotoxicity in HL60/ADR cells inhibiting the glycolysis by inactivating the Akt pathway.
牛磺酸上调基因1（TUG1）已被报道为急性髓系白血病（AML）中的一种致癌长链非编码RNA（lncRNA）。然而，TUG1在AML细胞耐药中的作用和分子机制仍不清楚。通过检测葡萄糖消耗和乳酸生成来评估糖酵解水平。采用qRT-PCR和蛋白质免疫印迹法检测TUG1、己糖激酶2（HK2）和丙酮酸激酶同工酶M2（PKM2）的表达。分别通过MTT法和流式细胞术评估阿霉素（ADR）的细胞毒性和细胞凋亡。通过对磷酸化Akt（p-Akt）（ser473）和Akt进行蛋白质免疫印迹分析来确定蛋白激酶B（Akt）信号通路的变化。我们的结果表明，HL60/ADR细胞中的糖酵解增加，这通过葡萄糖消耗和乳酸生成的增加以及HK2和PKM2在mRNA和蛋白质水平的表达增加得到证明。TUG1在HL60/ADR细胞中上调，敲低TUG可抑制糖酵解。敲低TUG1可增强ADR诱导的HL60/ADR细胞的细胞毒性和细胞凋亡。敲低TUG1可抑制Akt信号通路，而740Y-P激活Akt信号通路可减弱敲低TUG1对ADR诱导的细胞毒性、细胞凋亡以及HL60/ADR细胞糖酵解的影响。综上所述，敲低TUG1可增强阿霉素对HL60/ADR细胞的细胞毒性，通过使Akt信号通路失活来抑制糖酵解。
