SLC真核表达质粒的剂量-趋化效应及抑瘤效应研究

Study on dose-chemotactic and antitumor effects of SLC eukaryotic expression plasmid

目的:观察次级淋巴组织趋化因子(SLC)真核表达质粒剂量-趋化效应关系及不同剂量体内应用时对小鼠H22肝癌的治疗作用。方法:构建了小鼠SLC真核表达质粒(pSLC),体外转染检测pSLC在体外的剂量-趋化效应关系,体内不同剂量的pSLC质粒局部注射检测其表达情况及体内的剂量-趋化效应关系,并观察不同剂量的pSLC对小鼠H22肝癌治疗的剂效关系。结果:当剂量在0·5μg以下时,pSLC在体外转染的趋化效应与转染剂量呈正相关,但在0·75μg时明显下降;pSLC体内局部注射时,在所检测的200μg剂量以内,随注射质粒剂量的增加,其表达的mRNA水平逐渐增加,相应地,对淋巴细胞的趋化作用亦逐渐增强,pSLC对肿瘤的抑制作用亦随其剂量的增加而明显增强(P<0·001)。结论:体内局部转染表达pSLC,能够有效趋化聚集免疫细胞,对肿瘤产生免疫治疗效应。通过提高转染表达效果,可增加SLC趋化作用,增强肿瘤免疫治疗效应。

Objective： To investigate the dose-chemotactic effect relationship of secondary lymphoid tissue chemokine （SLC） eukaryotic expression plasmid and the dase-effect relationship of SLC plasmid on mouse H22 hepatocarcinoma. Methods： Mouse SLC eukaryotic expression plasmid（pSLC） was constructed. Dose-chemotaxis relationship of pSLC was determined by transfection of pSLC in vitro. SLC expression and dose-chemotaxis relationship of pSLC in vivo was observed after local injection with different dosage of pSLC plasmid. The therapeutical effect of different plasmid dosage on mouse H22 hepatocarcinoma was investigated. Results： Positive correlation of dose-chemotaxis relationship was observed when the transfection dosage of pSLC in vitro was below 0. 5 μg. However, the chemotactic response was obviously declined at dose 0. 75 μg. When plasmid was locally injected in vivo below the dose 200 μg, correspondingly to the increase of pSLC dosage, SLC mRNA expression level was increased, the chemotactic response of lymphocytes was augmented, and the therapeutical effect on H22 hepatocarcinoma was also obviously enhanced（P 〈 0. 001 ）. Conclusion： Antitumor immune effect could be obtained in the face of more immune cells chemoattracted by local tranofection of pSLC in vivo. The chemotactic effect could be enhanced by improving the transfection efficiency of pSLC, and thus the antitumor effect could be augmented.