摘要
【目的】提高阳离子脂质体介导的肺癌细胞基因转染效率。【方法】采用表达E .coliβ 半乳糖苷酶的 pCMVβ报告基因质粒及组化染色评价阳离子脂质体的转染效率。优化脂质体介导肺癌细胞基因转染效率的条件包括 :脂质体∶DNA电荷比率 ,DNA转染剂量 ,阳离子脂质体的类型。【结果】当阳离子脂质体∶DNA电荷比率 (+ / - )为 3∶1时 ,pCMVβ 的剂量为4μg/孔 (2 4孔板 )时 ,阳离子脂质体DOSPER介导肺癌细胞株SPC A 1转染效率最高 ,达 (11 9± 0 9) %。【结论】优化转染条件 ,脂质体可有效地将外源基因体外导入肺癌细胞。本实验为阳离子脂质体介导肺癌细胞功能性基因的转移及进一步的肺癌基因治疗打下了基础。
Objective This experiment aims at enhancing the efficiency of cationic liposome-mediated gene transfer to cultured lung cancer cells. Methods Reporter gene expressing E.coli β-galactosidase were used in order to evaluate transfection efficiency by histochemical staining. Optimization of conditions for cationic liposome-mediated transfection of cultured lung cancer cells include: charge ratios of cationic liposome: pCMVβ(+/-),doses of transferred pCMVβ, and the types of liposomes. Results When the cationic liposome: pCMVβ charge ratio is 3∶1 and dose D(Per well of 24-well plate)/ μg of pCMVβ is 4, the highest efficiency of DOSPER mediated gene transfer into SPC-A-1 cell reachs(11 9±0 9)%. Conclusions Optimising the conditions of transfection, cationic liposomes can effectively transfer the gene into cultured lung cancer cells.The results lay the fundations of functional gene transfer to lung cancer cells and further to the gene therapy of lung cancer.
出处
《中山医科大学学报》
CSCD
2000年第6期425-428,共4页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
在读博士研究生资助项目(331226)
作者简介
李芳(1965-),女,重庆人,博士生
