MMP-9 过表达对骨肉瘤143B 细胞 侵袭与迁移能力的影响

Effect of MMP-9 over-expression on invasion and migration of osteosarcoma 143B cells

摘要:

目的 研究MMP-9 过表达对成骨肉瘤143B 细胞侵袭与迁移能力的影响。方法 通过慢病毒介导 MMP-9 过表达载体转染骨肉瘤143B 细胞，实验分为3 组：实验组（转染pLV-Puro MMP-9 过表达序列）、 对照组（转染pLV-Puro MMP-9-NC 序列）、空白组（PBS）；实时荧光定量聚合酶链反应（qRT-PCR）检 测MMP-9 和VEGF mRNA 的表达；Western blot 法检测MMP-9 和VEGF 蛋白的表达；Transwell 小室侵袭 实验检测细胞侵袭能力；细胞划痕实验检测细胞迁移能力。结果 实验组MMP-9 和VEGF 的mRNA 和蛋 白表达量较对照组和空白组上调（P < 0.05）；实验组穿过人工基底膜细胞数高于对照组和空白组（P < 0.05）； 实验组划痕愈合率高于对照组和空白组（P <0.05）。结论 慢病毒介导MMP-9 过表达通过作用于VEGF 靶 基因，抑制成骨肉瘤143B 细胞侵袭及迁移能力。

Abstract:

Objective To study the effect of MMP-9 over-expression on invasion and migration of osteosarcoma 143B cells. Methods Lentivirus-mediated MMP-9 over-expression vector was transfected into osteosarcoma 143B cells. The experiment was divided into an experimental group (transfected with pLV-Puro MMP-9 over-expression sequence), a control group (transfected with pLV-Puro MMP-9-NC sequence) and a blank group (PBS). qRT-PCR was used to detect the expressions of MMP-9 and VEGF mRNAs. The expressions of MMP-9 and VEGF proteins were detected by Western blot. Transwell invasion assay was used to detect the invasion ability. Cell migration was measured by cell scratch test. Results The mRNA and protein expressions of MMP-9 and VEGF in the experimental group were significantly higher than those in the control and blank groups (P < 0.05). The number of cells passing through the artificial basement membrane in the experimental group was significantly larger than that in the control group and the blank group (P < 0.05). The healing rate of scratches in the experimental group was significantly higher than that in the control group and the blank group (P < 0.05). Conclusions Lentivirus-mediated MMP-9 overexpression inhibits the invasion and migration of osteosarcoma 143B cells by acting on VEGF target gene.