Targeting of human squamous carcinomas by SPA470-doxorubicin immunoconjugates.
Targeting of human squamous carcinomas by SPA470-doxorubicin immunoconjugates.
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SPA470-阿霉素免疫偶联物靶向人类鳞状细胞癌。
DOI：
10.1080/1061186031000121478
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发表时间：
2003
影响因子：
4.5
通讯作者：
Sauk,JJ
中科院分区：
文献类型：
--
作者：
Hebert,C;Norris,K;Sauk,JJ
在寻求确定头颈癌和鳞状细胞癌的有利靶点的过程中，我们试图确定Hsp 47/CBP 2是否可以用作靶点以及该靶点的表达是否受缺氧的影响。此外，我们确定了针对Hsp 47/CBP 2的多柔比星（DOX）免疫缀合物是否具有高细胞毒性药物活性和抗体定向杀伤携带抗原的肿瘤靶细胞，所述多柔比星（DOX）免疫缀合物将单克隆抗体（MAbs）连接到药物的13-酮位置。使用从美国典型培养物保藏中心（ATCC）（Manassas，VA）获得的人口腔鳞状细胞癌细胞（SCC）（SCC-4、SCC-9、SCC-15和SCC-25）的已建立细胞系进行实验。此外，UMB 2细胞系是SCC-9的自发突变体，其不表达Hsp 47/CBP 2。免疫缀合物的合成通过用2 IT硫醇化MAb并使MAb与DOX-腙反应来完成。通过间接免疫荧光测定MAb-DOX缀合物与SCC细胞的结合，并使用具有Cell Quest软件的Becton Dickinson FACS扫描进行分析。在常氧和缺氧期间，使用有限稀释测定和集落存活测定来确定DOX、MAb-DOX缀合物和MAb+DOX的细胞毒性的比较。这些研究显示，用SPA 470-DOX缀合物处理2小时的SCC细胞保留了对靶向SCC细胞的原始结合活性，并且比未缀合的DOX、DOX-腙或等效MAb蛋白+DOX显著更有效。而且，SPA 47-DOX产生的细胞杀伤力与等量的游离DOX相等，并且在较低浓度下比等量的游离DOX更大。在缺氧过程中，用SPA 470-DOX处理的细胞表现出集落存活率的小幅增加和细胞毒性的降低。SPA 470-DOX缀合物靶向表达Hsp 47/CBP 2的SCC细胞。SPA 470-D 0X在缺氧或模拟缺氧的条件下有效的证明假定了SPA 470-D 0X在治疗头颈癌中的进一步用途。
In a quest to identify a favorable target for head and neck cancers and squamous cell carcinoma, we sought to determine if Hsp47/CBP2 could be used as a target and whether the expression of this target was influenced by hypoxia. Moreover, we determined if doxorubicin (DOX) immunoconjugates directed against Hsp47/CBP2 that linked monoclonal antibodies (MAbs) to the 13-keto position of the drug possessed high cytotoxic drug activity and antibody-directed killing of antigen bearing tumor target cells. Experiments were performed using established cell lines of human oral squamous carcinoma cells (SCCs) (SCC-4, -9, -15 and -25) obtained from American Type Culture Collection (ATCC) (Manassas, VA). In addition, the UMB2 cell line is a spontaneous mutant of SCC-9 that does not express Hsp47/CBP2 was also used. Synthesis of the immunoconjugates was accomplished by thiolating the MAbs with 2 IT and reacting the MAbs with the DOX-hydrazone. The binding of MAb-DOX conjugates to SCC cells was determined by indirect immunofluorescence and analyzed using a Becton Dickinson FACS scan with Cell Quest software. Comparison of the cytotoxicity of DOX, MAb-DOX conjugates and MAb+DOX were determined using a limited dilution assay and colony survival assays during normoxia and hypoxia. These studies revealed that SCC cells treated with the SPA470-DOX conjugate for 2 h retained the original binding activity for targeted SCC cells and was significantly more potent that unconjugated DOX, DOX-hydrazone or equivalent MAb protein+DOX. Also, SPA47-DOX produced equal to and at lower concentrations greater cell killing than equivalent dose of free DOX. During hypoxia cells treated with SPA470-DOX demonstrated a small increase in colony survival and a diminishment in cytotoxicity. SPA470-DOX conjugates target SCC cells that express Hsp47/CBP2. The demonstration that SPA470-DOX is effective during hypoxia or conditions that mimic hypoxia presumes the further utility of SPA470-DOX in treating head and neck cancers.