合成肽VAR2特异性结合循环肿瘤细胞的检测及临床应用  

Detection and clinical application of VAR2 synthetic peptide specific binding to circulating tumor cells

作　　者：余宏 丁利杰 刘厚聪 王纪东 杜冀晖 YU Hong;DING Lijie;LIU Houcong;WANG Jidong;DU Jihui(Research Center for Clinical and Translational Medicine,Huazhong University of Science and Technology Union Shenzhen Hospital,Shenzhen 518052;Medical School,Shenzhen University,Shenzhen 518052,China)

机构地区：[1]华中科技大学协和深圳医院临床医学转化中心,广东深圳518052 [2]深圳大学医学部,广东深圳518052

出　　处：《基础医学与临床》2023年第11期1662-1667,共6页Basic and Clinical Medicine

基　　金：深圳市基础研究专项(自然科学基金)(JCYJ20190809105001757)。

摘　　要：目的探讨疟疾蛋白(VAR2)合成肽特异性结合循环肿瘤细胞(CTCs)的检测及临床应用。方法选择疟疾蛋白VAR2CSA功能结构域肽段序列合成VAR2多肽;基于纳米微流控技术富集CTCs,利用VAR2合成肽对富集细胞进行检测,建立一种新型CTCs检测方案。用细胞免疫荧光染色、流式细胞测量术及肿瘤细胞掺入回收实验,分析VAR2合成肽与不同组织来源肿瘤细胞特异性结合、回收效率,并与免疫荧光原位杂交(imFISH)检测法进行比较。选取22例结直肠癌患者与22名健康者,利用VAR2合成肽检测两者外周血CTCs。比较不同TNM分期患者CTCs阳性检出率。结果合成肽VAR2可以与人结肠癌细胞系SW620、人乳腺癌细胞系MCF7和人食管鳞癌细胞系KYSE180不同组织来源的肿瘤细胞特异性结合,合成肽VAR2检测100个SW620、MCF7、KYSE180肿瘤细胞掺入3 mL外周血中的回收率分别为82.8%±8.41%、56.2%±3.57%、86.6%±8.19%,与imFISH法检测法比较,差异无统计学意义。合成肽VAR2检测50、100、200个3种不同肿瘤细胞的回收率与掺入肿瘤细胞的数量没有明显相关性。合成肽VAR2检测临床结直肠癌患者外周血样本CTCs阳性率为63.6%,高于健康受试者组CTCs阳性率的0.0%,同时与侵袭深度、淋巴结转移、远处转移及临床分期相关,差异有统计学意义(P<0.05)。结论合成肽VAR2可实现对不同组织来源、非抗体依赖性的特异性CTCs检测,对于肿瘤患者临床分期、病情评估具有一定的临床应用价值。Objective To investigate the detection and clinical application studies based on malaria protein VAR2CSA(VAR2)synthetic peptide specifically binding to circulating tumor cells(CTCs).Methods The peptide sequence of the functional structural domain of malaria protein VAR2CSA was selected to synthesize VAR2 that enriched CTCs based on nano-microfluidic technology,and used VAR2 synthetic peptide to detect enriched cells to establish a novel detection protocol for CTCs.Cellular immuno-fluorescence staining,flow cytometry and tumor cell doping recovery assays were used to analyze the specific binding and recovery efficiency of VAR2 synthetic peptide with tumor cells of different tissue sources and compare with immuno-staining-fluorescence in situ hybridization(imFISH)assay.Twenty-two colorectal cancer patients and twenty-two healthy individuals were selected,and the VAR2 synthetic peptide was used to detect peripheral blood CTCs in both.The VAR2 synthetic peptide was used to compare the positive detection rate of CTCs in patients with different TNM stages.Results The VAR2 synthetic peptide bound specifically to tumor cells collected from different tissue sources of human colon cancer cell line SW620,human breast cancer cell line MCF7 and human esophageal squamous carcinoma cell line KYSE180.The VAR2 synthetic peptide detected 100 SW620,MCF7 and KYSE180 tumor cells mixed into 3 mL of peripheral blood with recovery rates of 82.8%±8.41%,56.2%±3.57%,86.6%±8.19%,which were not significantly different from the imFISH assay.The VAR2 synthetic peptide assay recoveries of 50,100,and 200 SW620 tumor cells were 80.7%,82.8%,and 84.7%,respectively,which did not correlate significantly with the number of adulterated tumor cells.The VAR2 synthetic peptide detected 63.6%positive CTCs in peripheral blood samples from patients with clinical colorectal cancer,which was higher than the 0.0%positive CTCs rate in the group of healthy subjects,and also correlated with depth of invasion,lymph node metastasis,distant metastasis,and clinical

关 键 词：循环肿瘤细胞 疟疾蛋白VAR2CSA(VAR2) 非抗体依赖性 结直肠癌 临床分期 

分 类 号：R446[医药卫生—诊断学]