稳定表达TSLC1基因骨肉瘤MG63细胞系的建立及鉴定

目的 建立稳定表达人肺癌抑癌基因-1（TSLC1）骨肉瘤细胞系并对其进行鉴定.方法 脂质体转染的方法 将真核表达载体pCI-TSLC1稳定转染至MG63细胞系,经克隆化培养以及G418筛选,获得了1株稳定高表达TSLC1蛋白的细胞系.对其表达的蛋白性质进行了鉴定,对其分泌动态、细胞纯度、遗传稳定性、外源基因整合等生物学特性做了鉴定.结果 获得高表达TSLC1的稳定细胞系,命名为M8T.生物学性状研究结果 表明该转基因细胞系纯度好,遗传性状稳定,抽提细胞RNA进RT-PCR,扩增到与预期结果 大小一致的DNA片段,说明TSLC1基因成功整合到细胞基因组中.细胞系无细菌和霉菌污染.结论 成功建立了稳定表达TSLC1的骨肉瘤细胞系M8T.该细胞系遗传性质稳定,为进一步研究抑癌基因TSLC1在骨肉瘤中的作用奠定了基础.

Establishment and identification of MG63 cell line stably expressing TSLC1

Objective To establish the osteosarcoma cell subline which stably expressing TSLC1 protein and to identify its biological characteristics. Methods The recombinant plasmid pCI-TSLC1 was stably transfected into MG63 cells with Lipofectamine 2000. The positive clones were developed by selection by G418. Biological characteristics of one of the 16 cell lines, namely, the MST were studied. Results The eukaryotic expression vector pCI-TSLC1 was successfully constructed and the stable cell subline highly expressing TSLC1 protein was obtained. The genetic stability and purity of the cell population were confirmed. The kinetics and genetics of MST were stable. The DNA product amplified from total RNA of osteosarcoma cell sublines revealed that ectogenous gene TSLC1 was integrated into the genomes of the MG63 cells. The cell line MST was not contaminated by microorganisms. Conclusion The stable osteosarcoma cell sublines highly expressing TSLC1 were successfully established, which provided a basis for further exploring the roles of TSLC1 in osteosarcoma.