运用蛋白质组学方法分析尤文肉瘤患者化疗前后血液差异表达蛋白的变化  

An analysis of differentially expressed proteins in blood in Ewing sarcoma patients before and after chemotherapy by proteomics

作　　者：张杰[1] 田征[1] 王翀[1] 宋兴华[1] 曹力[1] 艾克拜尔·尤努斯[1] 周义军[1] 吾斯曼·拜都拉 ZHANG Jie;TIAN Zheng;WANG Chong(Department of Bone Tumor Surgery,First Affiliated Hospital,Xinjiang Medical University,Urumqi 830000,CHINA)

机构地区：[1]新疆医科大学第一附属医院骨肿瘤外科,新疆乌鲁木齐830000

出　　处：《江苏医药》2021年第7期697-700,707,共5页Jiangsu Medical Journal

基　　金：新疆维吾尔自治区自然科学基金(2015211C094)。

摘　　要：目的运用蛋白质组学方法分析尤文肉瘤(ES)患者化疗前后全血样本差异表达蛋白的变化。方法收集4例ES患者化疗前(化疗前组)和化疗4个周期结束后(化疗后组)全血标本及4例自愿入组的健康人(健康组)的全血标本。ES患者均行4个周期长春新碱、阿霉素、环磷酰胺/异环磷酰胺及依托泊苷(VAC/IE方案)化疗。采用同位素标记相对和绝对定量技术联合液相串联质谱鉴定三组差异表达蛋白。Western blot技术验证三组超氧化物歧化酶1(SOD1)蛋白表达变化。结果与健康组比较,化疗前组差异表达蛋白显著上调。化疗后组相较于化疗前组下调的10种蛋白分别为:肌球蛋白轻链2、SOD1、胶原蛋白α1(Ⅰ)链、α辅肌动蛋白2、肌球蛋白轻链3、膜连蛋白A6、转胶蛋白、α1酸性糖蛋白1、酸性磷酸酶5型和胶原蛋白α2(Ⅵ)链。SOD1在化疗前组较健康组表达上调差异倍数为3.777倍,而化疗后组相较于化疗前组表达下调差异倍数为2.547倍。Western blot检测结果显示,与健康组相比,SOD1在化疗前组升高;而与化疗前组相比,化疗后组SOD1表达降低(P<0.05)。结论运用蛋白质组学分析可发现SOD1在尤文肉瘤中的特异性表达;SOD1表达增高可能在尤文肉瘤的发生、发展中发挥一定作用。Objective To analyze the differentially expressed proteins in blood in Ewing sarcoma(ES) patients before and after chemotherapy using proteomics analysis.Methods The whole blood samples were collected in 4 ES patients before chemotherapy(group A) and after 4 cycles of chemotherapy(group B) and in 4 healthy people(group C).ES patients received 4 cycles of vincristine, doxorubicin, cyclophosphamide/ifosfamide and etoposide chemotherapy(VAC/IE regimen).The isobaric tags for relative and absolute quantitation combined with liquid phase tandem mass spectrometry were used to identify differentially expressed proteins in three groups.Western blot technology was used to verify the protein expression of superoxide dismutase 1(SOD1) in three groups.Results Compared with group C,the differentially expressed proteins were up-regulated greatly in group A.Compared with group A,the down-regulated differentially expressed proteins in group B were myosin light chain 2,SOD1,collagen α1(Ⅰ) chain, α-actinin 2,myosin light chain 3,annexin A6,transfer protein, α1 acid glycoprotein 1,acid phosphatase type 5,and collagen α2(Ⅵ) chain.Compared with group C,the expression of SOD1 in group A was up-regulated by 3.777 times, which in group B was down-regulated by 2.547 times.Western blot results showed that the expression of SOD1 was more in group A than that of group C,which in group B was less than that in group A(P<0.05).Conclusion Proteomics analysis can be used to find the specific expression of SOD1 in blood in ES patients.The increased expression of SOD1 may play a role in the occurrence and development of ES.

关 键 词：尤文肉瘤 超氧化物歧化酶1 同位素标记相对和绝对定量技术 液相串联质谱分析 蛋白质免疫印迹 

分 类 号：R738[医药卫生—肿瘤]