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为了观察肿瘤坏死因子相关凋亡诱导配体(TRAIL)基因对体外培养的小鼠蜕膜基质细胞增殖及凋亡的作用,探讨TRAIL对小鼠子宫蜕膜化进程的影响,构建TRAIL过表达及干扰质粒,转染小鼠基质细胞后诱导蜕膜化发生.转染72h后,应用半定量RT-PCR和Western blotting检测蜕膜基质细胞中TRAILmRNA和蛋白质的表达情况、MTT法观察蜕膜基质细胞的生长和增殖能力、流式细胞术检测蜕膜基质细胞的细胞周期分布情况和凋亡率.经酶切和核苷酸测序证实,TRAIL基因正确克隆入真核表达载体且能够上调TRAIL的表达,干扰质粒能有效地抑制TRAIL基因的表达.TRAIL过表达和RNA干扰的结果表明:TRAIL具有将蜕膜基质细胞阻滞在G0/G1期、抑制蜕膜基质细胞增殖并促使其凋亡的功效,提示TRAIL可能参与调节胚胎植入后基质细胞的有序蜕膜化进程.
In order to observe the effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene on the proliferation and apoptosis of mouse decidual stromal cells cultured in vitro, the effect of TRAIL on the process of uterine decidualization and the construction of TRAIL overexpression and The expression of TRAIL mRNA and protein in decidual stromal cells was detected by semi-quantitative RT-PCR and Western blotting 72 h after transfection, and the expression of TRAIL mRNA and protein in decidual stromal cells was detected by MTT assay The cell cycle distribution and apoptosis rate of decidual stromal cells were detected by flow cytometry.It was confirmed by restriction enzyme digestion and nucleotide sequencing that the TRAIL gene was correctly cloned into the eukaryotic expression vector and could upregulate the expression of TRAIL , And the interference plasmids could effectively inhibit the expression of TRAIL gene.The results of TRAIL over-expression and RNA interference showed that TRAIL has the function of arresting decidual stromal cells in G0 / G1 phase, inhibiting the proliferation and promoting the apoptosis of decidual stromal cells , Suggesting that TRAIL may be involved in regulating the process of orderly decidualization of stromal cells after embryo implantation.