李纯璞;张立清;侯存强;郭冬梅;张月东;于鑫;

• 1:泰安市中心医院骨科
• 2:山东省泰安荣军医院
• 3:解放军960医院泰安院区骨科
• 4:泰安市中心医院血液内科

摘要(Abstract)：

[目的]检测乏氧和Notch通路相关蛋白在人骨肉瘤标本组织中的表达情况,并采用MG-63细胞实验研究多种相关蛋白参与骨肉瘤化疗的耐药机制。[方法]免疫组织化学方法检测HIF-1α、Notch1、MDR1/P-gp、MRP1蛋白在骨肉瘤及骨软骨瘤临床标本组织中表达情况。采用Notch si RNA转染乏氧环境中的MG-63细胞,Western blot检测相关蛋白表达情况,MTT方法检测细胞化疗药物敏感性的影响。[结果]骨肉瘤组织中HIF-1α、Notch1、MDR1/P-gp、MRP1蛋白的阳性表达率均显著高于骨软骨瘤组织,差异有统计学意义(P<0.05)。HIF-1α与Notch1之间,Notch1与MRP1之间,以及HIF-1α与MRP1之间存在显著正相关(P<0.05),而MDR1/P-gp与其他三种蛋白表达无相关性(P>0.05)。乏氧状态下MG-63细胞对顺铂、阿霉素、甲氨蝶呤或异环磷酰胺的半数抑制浓度IC50显著高于常氧状态,差异有统计学意义(P<0.05)。而乏氧环境Notch1 si RNA转染的MG-63细胞对顺铂、阿霉素、甲氨蝶呤或异环磷酰胺的半数抑制浓度IC50显著降低,差异有统计学意义(P<0.05),且Notch1-ICN和MRP1蛋白表达水平明显降低。[结论]骨肉瘤组织中HIF-1α、Notch1、MDR1/P-gp、MRP1蛋白的阳性表达率高于骨软骨瘤。乏氧条件下MG-63细胞对多种化疗药物呈耐药性,阻断Notch1可逆转乏氧诱导的骨肉瘤细胞多药耐药,提示Notch1是治疗骨肉瘤耐药的有效分子靶点。

关键词(KeyWords)： 骨肉瘤;乏氧;细胞耐药;多药耐药相关蛋白;Notch信号通路

Abstract:

Keywords:

基金项目(Foundation): 山东省医药卫生科技发展计划项目(编号:2016WS0595、2016WS0599、2018WS146);; 泰安市科技发展计划项目(编号:2016NS1055、2018NS0163)

作者(Author): 李纯璞;张立清;侯存强;郭冬梅;张月东;于鑫;

Email:

DOI:

参考文献(References)：

• [1]Efremov YR,Proskurina AS,Potter EA,et al.Cancer stem cells emergent nature of tumor emergency[J].Front Genet,2018,9544.
• [2]De Francesco EM,Maggiolini M,Musti AM.Crosstalk between Notch,HIF-1αand GPER in breast cancer EMT[J].Int J Mo Sci,2018,19(7):21-27.
• [3]Li Z,Zhu Q,Hu L,et al.Anterior gradient 2 is a binding stabilizer of hypoxia inducible factor-1αthat enhances Co Cl2-induced doxorubicin resistance in breast cancer cells[J].Cancer Sci2015,106(8):1041-1049.
• [4]Fischer C,Leithner K,Wohlkoenig C,et al.Panobinostat reduces hypoxia-induced cisplatin resistance of non-small cell lung carcinoma cells via HIF-1αdestabilization[J].Mol Cancer,2015,144.
• [5]Raninga PV,Di TG,Vuckovic S,et al.Trx R1 inhibition overcomes both hypoxia-induced and acquired bortezomib resistance in multiple myeloma through NF-кβinhibition[J].Cell Cycle.2016,15(4):559-572.
• [6]Arnason T,Harkness T.Development maintenance and reversal of multiple drug resistance:at the crossroads of TFPI1,ABC transporters and HIF1[J].Cancers(Basel),2015,7(4):2063-2082.
• [7]Pouyafar A,Zadi Heydarabad M,Aghdam SB,et al.Resveratrol potentially increased the tumoricidal effect of doxorubicin on SKOV3cancer stem cells in vitro[J].J Cell Biochem,2019,1:1-7.
• [8]Tsubaki M,Takeda T,Tomonari Y,et al.Overexpression of HIF-1αcontributes to melphalan resistance in multiple myeloma cells by activation of ERK1/2,Akt,and NF-κB[J].Lab Invest,2019,99(1):72-84.
• [9]Lin CW,Wang LK,Wang SP,et al.Corrigendum:daxx inhibits hypoxia-induced lung cancer cell metastasis by suppressing the HIF-1α/HDAC1/Slug axis[J].Nat Commun,2017,8:14502.
• [10]Liu B,Yan S,Qu L,et al.Celecoxib enhances anticancer effect of cisplatin and induces anoikis in osteosarcoma via PI3K/Akt pathway[J].Cancer Cell Int,2017,17:1.
• [11]Bocci F,Gearhart-Serna L,Boareto M,et al.Toward understanding cancer stem cell heterogeneity in the tumor microenvironment[J].Proc Natl Acad Sci USA,2019,116(1):148-157.
• [12]Hossain F,Sorrentino C,Ucar DA,et al.Notch signaling regulates mitochondrial metabolism and NF-κB activity in triple-negative breast cancer cells via IKKα-dependent non-canonical pathways[J].Front Oncol,2018,8:575.
• [13]Yang Z,Guo L,Liu D,et al.Acquisition of resistance to trastuzumab in gastric cancer cells is associated with activation of IL-6/STAT3/Jagged-1/Notch positive feedback loop[J].Oncotarget,2015,6(7):5072-5087.
• [14]Pandya K,Wyatt D,Gallagher B,et al.PKCαattenuates jagged-1-mediated notch signaling in Erb B-2-positive breast cancer to reverse trastuzumab resistance[J].Clin Cancer Res,2016,22(1):175-186.
• [15]Zou J,Li P,Lu F,et al.Notch1 is required for hypoxia-induced proliferation,invasion and chemoresistance of T-cell acute lymphoblastic leukemia cells[J].J Hematol Oncol,2013,6:3.