张 健,谢 涛,李恒元.自噬蛋白LC3B参与伏立诺他联合顺铂对骨肉瘤细胞增殖抑制及凋亡的研究[J].肿瘤学杂志,2018,24(5):443-448.
自噬蛋白LC3B参与伏立诺他联合顺铂对骨肉瘤细胞增殖抑制及凋亡的研究
Autophagy-related Protein LC3B Contribute to the Effects of Suberoylanilide Hydroxamic Acid Combined with Cisplatin on Proliferation and Apoptosis of Osteosarcoma Cells
投稿时间:2017-12-22  
DOI:10.11735/j.issn.1671-170X.2018.05.B007
中文关键词:  自噬  伏立诺他  骨肉瘤  细胞周期阻滞
英文关键词:autophagy  suberoylanilide hydroxamic acid  osteosarcoma  cell cycle arrest
基金项目:国家自然科学基金青年项目(81602355)
作者单位
张 健 杭州市第一人民医院南京医科大学附属杭州医院 
谢 涛 杭州市第一人民医院南京医科大学附属杭州医院 
李恒元 浙江大学医学院附属第二医院 
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中文摘要:
      摘 要:[目的] 探讨伏立诺他( suberoylanilide hydroxamic acid,SAHA)联合顺铂(cisplatin,CDDP)对骨肉瘤细胞的增殖抑制及其可能机制。[方法] 以骨肉瘤细胞系HOS为研究对象,通过MTS测定伏立诺他联合顺铂对骨肉瘤细胞增殖抑制的效果,运用流式细胞术(flow cytometry,FCM),透射电子显微技术(transmission electron microscope,TEM),蛋白质印迹技术(Western Blot)检测对细胞周期阻滞,自噬小体产生及凋亡与自噬相关蛋白的表达情况。[结果] 伏立诺他联合顺铂能够明显抑制骨肉瘤细胞HOS的增殖(P<0.05)。经伏立诺他联合顺铂(SAHA 1μmol/L+CDDP 4μmol/L)处理HOS细胞后,细胞周期阻滞于S期(P<0.01)。TEM结果表明联合处理组诱导了自噬小体的产生。Western Blot结果显示联合处理组使得HOS细胞LC3B蛋白的表达增加,与对照组相比差异有统计学意义(P<0.05)。在加入自噬抑制剂3-MA后,不仅降低LC3B的表达,而且Caspase3、PARP表达也降低,与对照组相比差异有统计学意义(均P<0.05)。 [结论] 伏立诺他联合顺铂通过细胞周期阻滞及诱导自噬的发生,从而发挥对骨肉瘤细胞增殖抑制及诱导凋亡的作用。
英文摘要:
      Abstract:[Objective] To explore the inhibitory effect of suberoylanilide hydroxamic acid(SAHA) combined with cisplatin(CDDP) on the proliferation of human osteosarcoma cells and the underlying mechanisms. [Methods] The osteosarcoma cell line HOS was used for test. The inhibitory effect of SAHA combined with CDDP on the proliferation of HOS cells was determined using the MTS kit. Cell cycle was monitored by flow cytometry. The presence of autophagosome was detected using transmission electron microscopy(TEM). Western-blotting was used to detect autophagy and apoptosis-related protein induced by these two drugs. The autophagy inhibitor 3-MA was added to confirm the phenomenon above. [Results] SAHA combined with CDDP could obviously inhibit the proliferation of HOS cells(P<0.05). The cell cycle was arrested at S phase after treatment with 1 μmol/L SAHA and 4 μmol/L CDDP(P<0.05). TEM demonstrated autophagosome,and Western blotting demonstrated increased expression of autophagy-related protein LC3B in HOS cells after the combination treatment(P<0.05). When the autophagy inhibitor 3-MA was added,the expression of LC3B,Caspase 3,and PARP was reduced(all P<0.05). [Conclusion] SAHA combined with CDDP inhibit proliferation and induce apoptosis of osteosarcoma cells through arresting cell cycle at S phase and inducing autophagy.
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