论文部分内容阅读
目的:合成人肿瘤转移相关基因(MTA1)的反义脱氧寡核苷酸,观察其转染后对人骨肉瘤MG-63细胞系中MTA1表达及骨肉瘤侵袭性的影响。方法:免疫细胞染色观察人工合成正义、反义及无意义MTA1基因片段转染人骨肉瘤细胞MG-63后,人肿瘤转移相关基因的表达;RT-PCR和蛋白质印迹法检测MTA1基因mRNA和蛋白质表达水平的变化;Boyden小室体外侵袭实验检测转染前后细胞侵袭力的变化。结果:反义寡核苷酸处理骨肉瘤细胞后,MTA1 mRNA的表达下降(吸光度之比为25.09±0.21),与正义链组、无意义链组和空白对照组(35.19±0.17、33.20±0.23和37.17±0.18)相比差异有统计学意义,P<0.05。Boy-den小室体外侵袭实验检测显示,反义寡核苷酸处理后骨肉瘤细胞的透膜能力明显下降。结论:MTA1同骨肉瘤的转移能力密切相关,反义寡核苷酸的转染可阻遏骨肉瘤细胞中MTA1的表达,并因此有可能限制骨肉瘤的侵袭和转移。
OBJECTIVE: To synthesize antisense oligodeoxynucleotides of human metastasis-associated gene (MTA1) and observe the effect of transfection on the expression of MTA1 and the invasiveness of osteosarcoma in human osteosarcoma MG-63 cell line. Methods: Immunohistochemical staining was used to observe the expression of MTA1 mRNA and protein in human osteosarcoma MG-63 cells after artificial synthetic sense, antisense and senseless MTA1 gene fragments were transfected into human osteosarcoma cell line MG-63. The expression of MTA1 mRNA and protein was detected by RT-PCR and Western blotting Level changes; Boyden chamber in vitro invasion assay to detect changes in invasiveness of cells before and after transfection. Results: The expression of MTA1 mRNA decreased (the ratio of absorbance was 25.09 ± 0.21) in osteosarcoma cells after antisense oligodeoxynucleotide treatment, which was significantly different from that in sense strand group, nonsense strand group and blank control group (35.19 ± 0.17, 33.20 ± 0.23 And 37.17 ± 0.18), the difference was statistically significant, P <0.05. Boy-den cell invasion assay in vitro showed that antisense oligonucleotide treatment of osteosarcoma cells significantly decreased the ability of the membrane. CONCLUSION: MTA1 is closely related to the metastasis of osteosarcoma. Transfection of antisense oligonucleotide can block the expression of MTA1 in osteosarcoma cells and therefore may limit the invasion and metastasis of osteosarcoma.