转染TNFα基因对胶质瘤细胞致瘤性的影响

StudyofCarcinogenesisofHumanGliomaCelsTransfectedwithTNFαGeneWangChen,ZhouLiangfu,ZhouFanmin,△ShenZhaozhong,△LuoJianming(DepartmentofNeurosurgery;HuashanHospital,ShanghaiMedicalUniversity,Shanghai200040,LaboratoryofMolecularBiology,TumorHospital,Shangh

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）ｐｇｍｌ（Ｐ＜０．０５），其生物活性达３２０．０ｕｍｌ。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。

Purpose To investigate growth inhibition of TNFα gene transfected human glioma cells. Methods We transduced the TNFα gene into human glioma cells(SHG-44)by the Molony murine leukemia virus-derived retroviral vector.The transfected clones selected by G418 were in vitro cultured for the study for growth inhibition.Nude mouse inoculation was to study the carcinogensis. Results Average levels of TNFα concentration in clones culture mediums determined by an enzyme-linked immunoassay method was (5 198.7±3 757.4) and (3 217.4±1 180.6)pgml ( P <0.05).The biological activity of TNFα in the supernatants detected by a bioassay with sensitive L929 cells was 320.0uml,and TNFα mRNA in the transfected cells was detected by RT-PCR.Introduction of hTNFα gene into SHG-44 glioma cells inhibited the growth characteristics.The subcutaneous injection of 1×10 6 TNFα expressing glioma cells into nude mice resulted in a reduced tumor progression in comparison to the controls.A mixed population of untransfected glioma cells consisting fo 10% TNFα- transfected tumor cells was inoculated S.C.into mice,regression of tumor mass occurred. Conclusions [WT10.BZ〗The TNFα gene transfected glioma cells had decreased the carcinogensis.