摘要
选用LacZ基因作为报告基因，用电穿孔法与pSV2neo质位共转化导入人肺巨细胞癌高转移细胞株，经G418及X－gal组化法双重筛检，获得LacZ基因表达阳性的细胞克隆。经X－gal组化染色，该克隆70％细胞蓝染，再经软琼脂培养后，形成的集落蓝染率约为65％，单个集落中的细胞蓝染率可达100％，在裸小鼠皮下或尾静脉接种这一LacZ基因表达阳性的克隆细胞后，在形成的肿瘤及肺内转移灶的冰冻切片中，X－gal染色可观察到蓝染的癌细胞。但由于细胞周期不同步、遗传稳定性以及表达调控因素的影响，则可能是本实验中部分细胞染色阴性的原因。我们的实验表明，LacZ基因有可能在肿瘤转移的研究中作为一种理想标志基因。
LacZ gene chosen as report gene together with pSV2neo was cotransfected by electroporation into highly metastatic cell-line of human lung giant cell carcinoma. Undergoing G418 and X-Gal double screening, cell clone expressing LacZ was obtained. By using X-Gal double screening, cell clone expressing LacZ was obtained. By using X-Gal staining, 70% of cells of that clone became blue in colour. 65% colonies of that cell clone in soft agar culture were blue after staining by X-gal. In the single blue colony 100% of cells were blue. Tumors and lung metastasis fool were observed after inoculation of cells expressing LacZ gene subcutaneouly or intravenously. By using X-Gal staining lots of tumor cells in the frozen section specimen of tumors as well as lung metastasis fooi were observed in blue staining. The reason for negative cells staining may be due to the influence of cell cycle asynchronization. genetic stability or expression regulation factors. The present results indicate that LacZ gene can be an ideal marker for tumor metastasis study.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
1998年第2期164-167,共4页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家自然科学基金!No.38970802
No.39770820
