特异性抗肝癌噬菌体单链抗体的应用研究

目的体外观察特异性抗肝癌(HCC)噬菌体单链抗体(ScFv)对HCC细胞生长和凋亡的影响.方法通过构建的特异性抗HCC噬菌体ScFv库在大肠杆菌HB2151中表达,制备游离ScFv,应用细胞培养、DNA凝胶电泳和流式细胞仪(FCM)技术检测抗体对HCC细胞生长和凋亡的影响.结果特异性抗HCC噬菌体ScFv处理的HCC细胞的生长速度明显减慢,克隆形成和附壁能力减弱,细胞存活率降低,DNA凝胶电泳可见典型凋亡梯状带;FCM检测发现抗体处理后HCC细胞凋亡率达32.6%,明显高于对照组(P＜0.01).结论应用噬菌体表面呈现技术和基因克隆技术构建的特异性抗HCC噬菌体ScFv具有抑制HCC细胞生长、促进HCC细胞凋亡的功能.

Application of phage display ScFv that against HCC specifically in vitro

Objective To observe influence of the ScFv library, which against to hepatocellular carcinoma(HCC) specifically, on HCC cell growth and apoptosis in vitro. Methods HCC cell as experimental group was treated with the supernatant of bacteria cultured with ScFv, HCC cell as control group was treated with the supernatant of bacteria cultured without ScFv, HCC cell as blank group was not treated with any supernatant. The HCC cell growth of the three groups was detected by counting the number and observing the characteristics of HCC cell cultured within seven days. The apoptosis of HCC cell was detected by means of DNA gel electrophoreses and FCM. Results The HCC cell of group one grew much slower than group two and group three, DNA ladder was observed by way of gel electrophoresis. Apoptosis detected by FCM occurred in 32.6% cells of group one, only 3.7% of group two and 2.8% of group three. Conclusion The phage display ScFv that against to HCC could inhibit HCC cells' growth and prompt HCC cells' apoptosis.$$$$