塞来昔布减低HL-60和HL-60A细胞活力、诱导凋亡及抑制自噬
发布时间：2018-08-19 11:44
【摘要】：目的:探讨塞来昔布对急性髓细胞白血病(AML)HL-60细胞和HL-60A细胞的活力、凋亡及自噬的影响。方法:用不同浓度的(0、20、40、60、80和100μmol/L)塞来昔布作用于HL-60细胞和HL-60A细胞,24h、48h和72h后用MTT法检测细胞活力。用流式细胞术检测塞来昔布作用HL-60细胞和HL-60A细胞24 h后的凋亡率。用Western blot法检测凋亡相关蛋白cleaved caspase-3、cleaved PARP,自噬相关蛋白LC3、P62,以及mTOR信号途径相关蛋白。结果:塞来昔布作用于HL-60细胞24h、48h和72h的IC_(50)分别为49.4μmol/L、32.0μmol/L和25.1μmol/L,对于HL-60A细胞,相应的IC_(50)分别是69.1μmol/L、42.5μmol/L和29.6μmol/L。塞来昔布作用24h后,流式细胞术检测显示HL-60细胞中Annexin-V~+PI~-、Annexin-V~+PI~+及Annexin-V~-PI~+细胞的比例增多;HL-60A细胞中Annexin-V~+PI~-及Annexin-V~+PI~+细胞的比例增多。Western blot实验结果显示塞来昔布作用后,cleaved caspase-3和cleaved PARP的蛋白水平增高,提示该凋亡作用是通过caspase途径的。自噬相关蛋白LC3Ⅱ及P62的表达均增加,mTOR、p-mTOR以及下游的4-EBP、p-4-EBP的蛋白水平没有变化,说明塞来昔布能够抑制AML细胞自噬,该作用与mTOR途径无关。结论:塞来昔布对HL-60细胞和HL-60A细胞活力的抑制作用呈浓度以及时间依赖性,该作用与塞来昔布诱导细胞凋亡及坏死有关。塞来昔布能够通过非mTOR依赖途径抑制AML细胞自噬,有望联合应用于AML的治疗,有助于增强某些引起保护性自噬的化疗药物的细胞毒作用。
[Abstract]:AIM: To investigate the effect of celecoxib on the viability, apoptosis and autophagy of HL-60 cells and HL-60A cells in acute myeloid leukemia (AML). Methods: HL-60 cells and HL-60A cells were treated with celecoxib at different concentrations (0,20,40,60,80 and 100 micromol/L). Cell viability was measured by MTT at 24, 48 and 72 hours after treatment. The apoptosis rates of HL-60 cells and HL-60A cells were measured by Western blot. Apoptosis-related proteins cleaved caspase-3, cleaved PARP, autophagy-related proteins LC3, P62 and mTOR signaling pathway-related proteins were detected. Results: The IC_ (50) of HL-60 cells treated with celecoxib for 24 h, 48 h and 72 h were 49.4, 32.0 and 25.1 micromol/L, respectively. Flow cytometry showed that the proportion of Annexin-V~+PI~-, Annexin-V~+PI~+ and Annexin-V~+PI~+ cells increased in HL-60A cells, and the proportion of Annexin-V~+PI~- and Annexin-V~-PI~+ cells increased in HL-60A cells. The results showed that cleaved caspase-3 and cleaved PARP protein levels increased after celecoxib treatment, suggesting that the apoptosis was mediated by caspase pathway. The expressions of autophagy-associated proteins LC3 II and P62 increased, while the levels of mTOR, p-mTOR and downstream 4-EBP, p-4-EBP protein did not change, indicating that celecoxib could inhibit AML cells from autophagy. Conclusion: Celecoxib inhibits the activity of HL-60 cells and HL-60A cells in a concentration-and time-dependent manner, which is related to celecoxib-induced apoptosis and necrosis. Enhance the cytotoxicity of some chemotherapeutic drugs that cause protective autophagy.
【作者单位】： 中山大学附属第三医院输血科;中山大学附属第三医院血液科;
【基金】：广东省自然科学基金资助项目(No.2014A030313138);广东省自然科学基金博士启动基金资助项目(No.2014A030310292)
【分类号】：R733.7
，
本文编号：2191558
[Abstract]:AIM: To investigate the effect of celecoxib on the viability, apoptosis and autophagy of HL-60 cells and HL-60A cells in acute myeloid leukemia (AML). Methods: HL-60 cells and HL-60A cells were treated with celecoxib at different concentrations (0,20,40,60,80 and 100 micromol/L). Cell viability was measured by MTT at 24, 48 and 72 hours after treatment. The apoptosis rates of HL-60 cells and HL-60A cells were measured by Western blot. Apoptosis-related proteins cleaved caspase-3, cleaved PARP, autophagy-related proteins LC3, P62 and mTOR signaling pathway-related proteins were detected. Results: The IC_ (50) of HL-60 cells treated with celecoxib for 24 h, 48 h and 72 h were 49.4, 32.0 and 25.1 micromol/L, respectively. Flow cytometry showed that the proportion of Annexin-V~+PI~-, Annexin-V~+PI~+ and Annexin-V~+PI~+ cells increased in HL-60A cells, and the proportion of Annexin-V~+PI~- and Annexin-V~-PI~+ cells increased in HL-60A cells. The results showed that cleaved caspase-3 and cleaved PARP protein levels increased after celecoxib treatment, suggesting that the apoptosis was mediated by caspase pathway. The expressions of autophagy-associated proteins LC3 II and P62 increased, while the levels of mTOR, p-mTOR and downstream 4-EBP, p-4-EBP protein did not change, indicating that celecoxib could inhibit AML cells from autophagy. Conclusion: Celecoxib inhibits the activity of HL-60 cells and HL-60A cells in a concentration-and time-dependent manner, which is related to celecoxib-induced apoptosis and necrosis. Enhance the cytotoxicity of some chemotherapeutic drugs that cause protective autophagy.
【作者单位】： 中山大学附属第三医院输血科;中山大学附属第三医院血液科;
【基金】：广东省自然科学基金资助项目(No.2014A030313138);广东省自然科学基金博士启动基金资助项目(No.2014A030310292)
【分类号】：R733.7
，
本文编号：2191558
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2191558.html
最近更新
教材专著
