论文部分内容阅读
研究转染肿瘤坏死因子α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒα，ＴＮＦα）基因的胶质瘤细胞的致瘤性。方法运用ＭｏＭＬＶ逆转录病毒载体，将ＴＮＦα基因转导至人胶质瘤细胞ＳＨＧ－４４，用Ｇ４１８筛选出阳性细胞克隆，以细胞培养检测细胞生长抑制，以裸鼠接种研究其致瘤性。结果转染细胞培养上清液中ＴＮＦα含量分别为（５１９８．７±３７５７．４）和（３２１７．４±１１８０．６）ｐｇｍｌ（Ｐ＜０．０５），其生物活性达３２０．０ｕｍｌ。运用ＲＴ－ＰＣＲ检测ＴＮＦαｍＲＮＡ特异性表达。细胞培养发现：转染ＴＮＦα基因的细胞生长受抑制，Ｇ２－Ｍ期缩短而Ｇ０－Ｇ１期延长。裸鼠接种试验发现转染ＴＮＦα基因的胶质瘤细胞致瘤性下降，早期出现明显坏死。未转染ＴＮＦα基因的胶质瘤细胞混有１０％的转染ＴＮＦα基因的细胞，其生长也受到抑制。结论转染ＴＮＦα基因的胶质瘤细胞的致瘤性下降。
To study the tumorigenicity of glioma cells transfected with tumor necrosis factor α (TNFα) gene. Methods TNFα gene was transduced into human glioma cells SHG-44 using MoMLV retrovirus vector. Positive cell clones were screened out with G418. Cell growth inhibition was detected by cell culture. The tumorigenicity was studied by nude mice inoculation. Results The content of TNFα in transfected cell culture supernatant was (5198.7±3757.4) and (3217.4±1180.6) pgml (P<0.05), and its biological activity reached 320.0u. ml. The specific expression of TNFα mRNA was detected by RT-PCR. In cell culture, the growth of cells transfected with TNFα gene was inhibited, G2-M phase was shortened and G0-G1 phase was prolonged. Nude mouse inoculation test found that the tumorigenicity of glioma cells transfected with TNFα gene was decreased, and necrosis appeared early. Glioma cells not transfected with the TNFα gene were mixed with 10% of cells transfected with the TNFα gene, and their growth was also inhibited. Conclusion The tumorigenicity of glioma cells transfected with TNFα gene is decreased.