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[使用聚阳离子脂质体介导转导对小鼠头颈部鳞状细胞癌进行白细胞介素-2基因治疗的研究]

[Studies of mouse interleukin-2 gene therapy for head, and neck sequamous cell carcinoma using polycationic liposome-mediated transduction].

作者信息

Yang Huan, Liu Shixi, Liang Chuanyu, Peng Wenzhen

机构信息

Department of Otolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2003 Jan;34(1):9-11, 30.

Abstract

OBJECTIVE

To investigate the immunological mechanism of mouse IL-2 gene therapy and the optimal lipid to DNA ratios of lipid-DNA complexed (lipoplexes) by using polycationic liposome-mediated Tumors were established in the transduction for head and neck squamous cell carcinoma (HNSCC).

METHODS

floor of mouth in C3H/HeJ immunocompetent mice with SCCVII cell line. Lipoplexes with various lipid to DNA ratios (L:D) and naked DNA were transducted in vivo by direct intratumoral gene transfer and in vitro. The supernatants of SCCVII cell and tumour tissues were collected for IL-2 expression by enzyme-linked immunosorbent assay. Natural killer (NK) cell activity and cytotoxic T-lymphocyte (CTL) activity were also The optimal L:D ratio for IL-2 expression in vitro was not assayed by lactate dehydrogenase method.

RESULTS

consistent with that in vivo. By use of lipoplexes with L:D = 3:1, higher IL-2 expression of tumor tissues and greater activities of NK cell and CTL of murine spleen were noted in the treated group as compared with those A comparison of naked plasmid and lipid-complexed found in naked DNA and empty plasmid (EP).

CONCLUSION

DNA revealed that lipoplexes were more effective for intratumoral gene transfer to HNSCC. The results indicate that the formulation and dosage of polycationic L:D complexes play a key role in determining the level of intratumoral transgene expression.

摘要

目的

通过使用聚阳离子脂质体介导的转导来研究小鼠白细胞介素-2(IL-2)基因治疗的免疫机制以及脂质-DNA复合物(脂质体)的最佳脂质与DNA比例,用于头颈部鳞状细胞癌(HNSCC)的肿瘤治疗。

方法

用SCCVII细胞系在C3H/HeJ免疫活性小鼠的口腔底部建立肿瘤。通过直接瘤内基因转移在体内和体外转导具有不同脂质与DNA比例(L:D)的脂质体和裸DNA。收集SCCVII细胞和肿瘤组织的上清液,通过酶联免疫吸附测定法检测IL-2表达。自然杀伤(NK)细胞活性和细胞毒性T淋巴细胞(CTL)活性也通过乳酸脱氢酶法进行检测。

结果

体外IL-2表达的最佳L:D比例与体内一致。与裸DNA和空质粒(EP)相比,使用L:D = 3:1的脂质体时,治疗组肿瘤组织中IL-2表达更高,小鼠脾脏的NK细胞和CTL活性更强。

结论

裸质粒与脂质复合DNA的比较表明,脂质体对HNSCC的瘤内基因转移更有效。结果表明,聚阳离子L:D复合物的配方和剂量在决定瘤内转基因表达水平方面起关键作用。

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