反义硫代寡核苷酸对人脑胶质瘤细胞系转化生长因子α基因表达及生长抑制作用的研究

Antisense oligodeoxynucleotide of TGFα inhibits its gene expression and proliferation of a human glioma cell line

目的研究转化生长因子α（ＴＧＦα）反义寡核苷酸对人脑胶质瘤细胞系中该ＴＧＦα基因表达的作用。方法采用人工合成反义、正义及随机三组硫代脱氧寡核苷酸（ＳＯＮ）经阳性脂质体包裹后转染人脑胶质瘤ＴＪ９０５细胞系。用细胞计数检测转染后瘤细胞的生长抑制率，用细胞原位ＲＮＡ杂交与免疫组化方法检测ＴＧＦαｍＲＮＡ及蛋白的表达水平。结果ＴＧＦα反义ＳＯＮ可抑制ＴＪ９０５细胞系的生长和ＴＧＦαｍＲＮＡ及其蛋白的表达。结论ＴＧＦα反义ＳＯＮ能够抑制人脑胶质瘤ＴＪ９０５细胞系ＴＧＦαｍＲＮＡ及其蛋白的表达，同时细胞的生长受到明显抑制。

Objective To study the effect of antisense oligodeoxynucleotide(ON) of TGFα on its gene expression in TJ 905 cell line. Methods Antisense and sense TGFα phosphorothioate ON (SON) and random SON were synthesized and transfected to TJ 905 cells mediated by lipofectin. Their effects on the TGFα gene expression were examined by in situ hybridization of TGFα mRNA, immunohistochemical study and cell count. Results (1) TGFα antisense SON could significantly inhibit the growth of TJ 905 cell lines. The inhibition peaked at 24 hour after transfection, the inhibition rate reached 68% at 20 μM, and the effect decreased 72 hours after transfection. The inhibition effect was dose dependent. (2) Antisense TGFα SON inhibited TGFα expressions at both mRNA and protein levels. Conclusion Antisense TGFα SON can inhibit the expressions of TGFα and markedly inhibit cell growth of human glioma TJ 905 cell line. TGFα contributes in the growth potential of glioma.