摘要
目的扩增纯化携带融合基因SLC-Her-2/neu-p53-Fc(SLC-HP-Fc)的重组腺病毒AdEasyTM-SLC-HP-Fc真核表达载体,研究其对荷黑色素瘤小鼠的免疫治疗效果。方法将重组腺病毒真核表达载体AdEasyTM-SLC-HP-Fc在人胚肾293细胞中扩增、纯化;向小鼠皮下注射表达Her-2/neu-p53融合基因的B16F10-psig-Her-2/neu-p53细胞(B16-HP细胞)建立荷黑色素瘤小鼠模型;将小鼠随机分成对照组、肌肉注射组和皮下注射组,用重组腺病毒AdEasyTM-SLC-HP-Fc真核表达载体进行治疗,观察肿瘤生长情况,测定细胞毒性活性、血清p53抗体。结果纯化后重组腺病毒AdEasyTM-SLC-HP-Fc真核表达载体滴度8×108pfu/mL。目的基因SLC-HP在小鼠体内成功表达。重组腺病毒AdEasyTM-SLC-HP-Fc真核表达载体高剂量肌肉注射能有效抑制小鼠肿瘤的生长,瘤重与对照组比较差异有统计学意义(P=0.005),抑瘤率达到98.8%,细胞毒性T淋巴细胞(CTL)的特异性杀伤力有明显提高。结论重组腺病毒AdEasyTM-SLC-HP-Fc真核表达载体高剂量肌肉注射能延长出瘤时间,抑制肿瘤的生长。
Objective To amplify and purify the recombinant adenovirus with fused gene of SLC, Her-2/neu, p53 and Fc fragment （SLC-HP-Fc） and test its immunotherapeutic effect on mice bearing melanoma. Methods The adenovirus was given to the C57BL/6 mice model bearing melanoma to test its anti-tumor effect. Results By LDH assay, we found the cytolysis activity of mice spleen lymphocytes from the mice treated with high dosage of AdEasyTM-SLC-HP-Fc by intramuscular injection was higher than that of the spleen lymphocytes of the mice from other groups. Oonclusion The fused gene transmission by the adenovirus vector is feasible. And the eukaryotic expression of the recombinant aderlovirus AdEasyTM-SLC-HP-Fc, especially with high dosage by intramuscular injection, can delay tumor emergence and inhibit tumor growth.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2014年第6期790-794,815,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
