目的探讨PIAS3表达对人脑胶质瘤TJ905细胞侵袭力的影响及其可能的机制。方法构建PIAS3过表达载体及合成PIAS3siRNA,转染TJ905细胞,上调或下调TJ905细胞中PIAS3表达水平,Transwell试验检测TJ905细胞的侵袭力,Westernblot验证PIAS3表达及基质金属蛋白酶抑制因子(TIMP)3、基质金属蛋白酶(MMP)-2、MMP-9蛋白的表达。结果体外转染质粒和寡聚核苷酸效率分别为85.3%±3.1%和95.1%±2.9%。体外转染PIAS3过表达质粒能有效提高TJ905细胞中PIAS3蛋白的表达,明显抑制TJ905细胞侵袭力(P〈0.05),细胞穿过率由对照组87.9%±9.3%降为37.3%±7.9%,同时上调TIMP3和下调MMP-2、MMP-9蛋白的表达(P〈0.05);转染PIAS3siRNA能有效抑制TJ905细胞中PIAS3蛋白的表达,增强TJ905细胞侵袭力(P〈0.05),细胞穿过率由对照组83.9%±7.1%增加到93.2%±3.1%,同时下调TIMP3和上调MMP-2、MMP-9蛋白的表达(P〈0.05)。结论PIAS3表达水平与胶质瘤TJ905细胞的侵袭特性密切相关。
Objectives To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells. Methods PIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or downregulation of PIAS3 expression levels in TJ905 cells. After that, the invasive effects of TJ905 cells were measured by Transwcll assay, and the expression of PIAS3, tissue inhibitor of metalloproteinases (TIMP)3, matrix metalloprotease (MMP)-2, and MMP-9 were identified by Western blot. Results In vitro transfection efficiency of plasmids and oligonucleotides were separately 85.3% ± 3. 1% and 95. 1% ± 2. 9%. PIAS3 overexpression plasmid transfection in vitro could effectively improve the expression of PIAS3 protein in TJ905 cells and inhibit the invasion of TJ905 cells (P 〈 0. 05), and cell penetration ratio reduced from 87. 9% ±9. 3% to 37.3%±7.9% compared with control group, while it upregulated TIMP3 and downregulated MMP-2, MMP-9 protein expression (P 〈 0. 05) ; PIAS3 siRNA transfection could inhibit the PIAS3 protein expression of TJ905 ceils and promote the invasion of TJ905 cells ( P 〈 0. 05 ), and cell penetration ratio increased from 83.9% ± 7. 1% to 93.2% ± 3.1% compared with control group, while it downregulated TIMP3 and upregulated MMP-2, MMP-9 protein expression (P 〈 0. 05 ). Conclusion PIAS3 expression is closely related to the invasion properties of glioma TJ905 cells.