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目的改进、优化体内噬菌体展示技术,为肝癌肿瘤血管异质性研究提供稳定、高效的技术平台。方法将激光捕获显微切割(LCM)与实时聚合酶链反应(Real-time PCR)技术与噬菌体文库体内展示相结合,在裸鼠肝癌原位种植模型上筛选与肝癌肿瘤血管内皮细胞特异结合的靶向噬菌体。结果应用LCM及Real-time PCR技术,不仅成功克服了肝脏对噬菌体文库的非特异性结合,肿瘤特异性噬菌体LCI-X7被成功富集,其与肿瘤血管的亲和力分别是肝、肺、肾、脑组织的12、33、426及745倍。结论经过改进优化的体内噬菌体展示技术可为肝癌肿瘤血管异质性研究提供稳定、高效的技术平台。
Objective To improve and optimize phage display in vivo and provide a stable and efficient technology platform for the study of tumor angiogenesis in liver cancer. Methods Laser capture microdissection (LCM) and Real-time PCR (Real-time PCR) combined with in vivo display of phage display library were used to screen the hepatocarcinoma tumor vascular endothelial cells Targeted phage. Results LCM and Real-time PCR not only successfully overcome the non-specific binding of liver to phage library, but also enrich the tumor-specific phage LCI-X7. The affinity of tumor-specific LCI-X7 is liver, lung, kidney and brain 12,33,426 and 745 times of tissue. Conclusion The improved and optimized in vivo phage display technology can provide a stable and efficient technology platform for the study of hepatic tumor vascular heterogeneity.