【摘要】 目的:评估sTRAIL基因引起人胚胎干细胞[human embryonic stem(hES)cells]和人恶性畸胎瘤(human malignant teratoma)细胞NTERA-2凋亡的作用。方法:本室已构建的pAAV-PEG3-sTRAIL质粒,转染NTERA-2和hES细胞,用流式细胞仪和定量PCR检测NTERA-2和hES细胞的早期凋亡及sTRAIL基因的受体的表达。结果:NTERA-2细胞经瞬时转染pAAV-PEG3-sTRAIL质粒后,早期凋亡比例达24.4%±2.1%,sTRAIL的死亡受体DR4、和DR5的表达分别上升2和3倍,而hES细胞转染pAAV-PEG3-sTRAIL质粒后,早期凋亡比例仅2.3%±1.2%,受体DR4、DR5、DcR1和DcR2的表达未有明显变化。结论:pAAV-PEG3-sTRAIL质粒可有效地引起人恶性畸胎瘤细胞NTERA-2的凋亡,但是对人胚胎干细胞尚未发现可见的细胞凋亡现象。更多还原

【Abstract】 Objective: To evaluate the apoptotic effects of human malignant teratoma(hMT) cells and human embryonic stem (hES) cells induced by sTRAIL gene. Methods: The NTERA-2 and hES cells were transiently transfected with pAAV-PEG3-sTRAIL plasmid constructed in our lab. FACS, real-time PCR, and semi-quantitative RT-PCR were employed to detect the early cell apoptosis and the expressions of sTRAIL gene and its receptors in NTERA-2 and hES cells treated with pAAV-PEG3-sTRAIL plasmid, respectively. Results: After NTERA-2 or hES cells were treated with pAAV-PEG3-sTRAIL plasmid, the early apoptotic cells accounted for 24.4% ± 2.1% or 2.3%±1.2%.Transfection with pAAV-PEG3-sTRAIL induced a>2-fold increase in DR4 expression and a >3-fold increase in DR5 expression, respectively, no significant change was found in the expression of either DcR1 or DcR2 in NTERA-2 cells. However, the expressions of DcR1, DcR2, DR4 and DR5 had no significant change in hES cells treated with pAAV-PEG3-sTRAIL.Conclusions: The plasmid pAAV-PEG3-sTRAIL has the ability to induce the cell apoptosis of NTERA-2 cells and no effect has been found on hES cells.更多还原