Xiao Min, Hong Zhenya, Sun Lishi, Wu Ying, Zhang Na, Liu Yanan, Luo Danfeng, Zhou Jianfeng, Li Chunrui
Department of Hematology, Huazhong University of Science and Technology, Wuhan, 430030, China.
Department of Hematology, Hospital Affiliated to Medical College, Qingdao University, Qindao, 266003, China.
J Huazhong Univ Sci Technolog Med Sci. 2011 Oct;31(5):608. doi: 10.1007/s11596-011-0569-y. Epub 2011 Oct 25.
TMTP1, a 5-amino acid peptide NVVRQ, obtained by using the flagella peptide library screening in our previous studies, can be used for the labeling of malignant in situ and metastatic lesions, and even micro-metastases. In this study, TMTP1 was assessed for its ability to specifically target the malignant hematopoietic cells and metastatic lesions of hematological malignancies. FITC-TMTP1 was chemically synthesized. Immunofluorescence assay and competitive test were carried out to determine the specific binding capacity of TMTPl to hematological malignant cell lines, including HL60, k562, SHI-1, Jurkat, Raji, El-4 and umbilical cord blood mononuclear cells. Mononuclear cells were isolated from the bone marrow of healthy subjects and patients with chronic myeloid leukemia. Then the cells were co-cultured with TMTP1 or scrambled peptides and the binding and affinity of TMTP1 peptide to the primary cells of hematological malignancies were flow cytometrically analyzed. The binding specificity of TMTP1 to target hematological malignancies was measured in vivo by intravenous injection of FITC-conjugated TMTP1 into El-4 lymphoma-bearing mice. The results showed that TMTP1 specifically bound to the cells of a series of hematological malignancies, including HL60, k562, Jurkat, Raji, El-4 and chronic myeloid leukemia primary cells but not to bone marrow mononuclear cells from healthy subjects. By contrast, TMTP1 could bind to the metastatic foci of lymphoma originating from the EL-4 cell line while the scrambled peptide failed to do so. Moreover, the occult metastases could be identified, with high specificity, by detecting FITC-TMTP1. We are led to conclude that TMTP1, as a novel tumor-homing peptide, can serve as a marker for primary malignant and metastatic lesions for the early diagnosis of hematological malignances and a carrier of anticancer drugs for cancer treatment.
TMTP1是一种由5个氨基酸组成的肽NVVRQ，在我们之前的研究中通过鞭毛肽库筛选获得，可用于标记原位恶性病变和转移灶，甚至微转移灶。在本研究中，评估了TMTP1特异性靶向血液系统恶性肿瘤的恶性造血细胞和转移灶的能力。化学合成了FITC-TMTP1。进行免疫荧光测定和竞争试验，以确定TMTP1与血液系统恶性细胞系（包括HL60、k562、SHI-1、Jurkat、Raji、El-4）以及脐带血单个核细胞的特异性结合能力。从健康受试者和慢性髓性白血病患者的骨髓中分离单个核细胞。然后将细胞与TMTP1或乱序肽共培养，并通过流式细胞术分析TMTP1肽与血液系统恶性肿瘤原代细胞的结合和亲和力。通过向荷El-4淋巴瘤的小鼠静脉注射FITC偶联的TMTP1，在体内测量TMTP1对靶向血液系统恶性肿瘤的结合特异性。结果表明，TMTP1特异性结合一系列血液系统恶性肿瘤的细胞，包括HL60、k562、Jurkat、Raji、El-4和慢性髓性白血病原代细胞，但不结合健康受试者的骨髓单个核细胞。相比之下，TMTP1可以结合源自El-4细胞系的淋巴瘤转移灶，而乱序肽则不能。此外，通过检测FITC-TMTP1可以高特异性地识别隐匿性转移灶。我们得出结论，TMTP1作为一种新型的肿瘤归巢肽，可作为原发性恶性病变和转移灶的标志物用于血液系统恶性肿瘤的早期诊断，也可作为抗癌药物载体用于癌症治疗。
