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目的研究转化生长因子(TGF)-β1反义寡核苷酸(ASON)对人胚胎性横纹肌肉瘤细胞系RD增殖的影响。方法采用反义寡核苷酸技术,以阳离子脂质体为载体,将人工合成的TGF-β1 ASON转染RD细胞,阻碍内源性TGF-β1/Smad信号的转导,以未转染的RD细胞组作对照。免疫荧光技术检测RD细胞TGF-β1蛋白表达水平;四甲基偶氮唑盐实验(MTT)检测RD细胞的增殖情况;流式细胞术(FCM)检测细胞周期。结果转染4 d后RD细胞TGF-β1蛋白表达量较转入2 d时明显减少,与对照组比较,差异有统计学意义(P<0.05);RD细胞活力明显降低,合成期(S期)细胞减少。结论TGF-β1 ASON可有效阻碍TGF-β1/Smad信号通路,在一定程度上可促进RD细胞退出细胞周期,并抑制RD细胞增殖。
Objective To investigate the effect of transforming growth factor-β1 antisense oligonucleotide (ASON) on RD proliferation in human embryonic rhabdomyosarcoma cell line. Methods The antisense oligonucleotide technique was used to transfect the synthetic TGF-β1 ASON into RD cells with cationic liposome as a vector to block the transduction of endogenous TGF-β1 / Smad signal. RD cell group as a control. The expression of TGF-β1 in RD cells was detected by immunofluorescence. The proliferation of RD cells was detected by MTT assay. The cell cycle was detected by flow cytometry (FCM). Results Compared with control group, the expression of TGF-β1 protein in RD cells was significantly decreased 4 days after transfection (P <0.05), while the viability of RD cells was significantly decreased Cells are reduced. Conclusion TGF-β1 ASON can effectively block the TGF-β1 / Smad signaling pathway, to a certain extent, promote the RD cell exit the cell cycle and inhibit the proliferation of RD cells.