基质金属蛋白酶-9对骨肉瘤细胞可溶性MICA蛋白形成的影响

Effect of matrix metalloproteinase-9 on the formation of soluble MICA protein in osteosarcoma cells

目的:探讨基质金属蛋白酶-9(MMP-9)对骨肉瘤细胞可溶性MICA(s MICA)蛋白形成的影响,为骨肉瘤免疫治疗提供新靶点。方法:脂质体法转染MMP-9反义寡核苷酸到骨肉瘤细胞U2OS,应用RT-PCR、明胶酶谱法和ELISA检测转染寡核苷酸后U2OS细胞MMP-9 m RNA表达,以及培养上清液中MMP-9的酶活性和s MICA蛋白浓度变化。结果:U2OS细胞转染MMP-9反义寡核苷酸后,MMP-9 m RNA表达及酶活性受到抑制;培养上清液中s MICA蛋白浓度下降(P=0.011)。结论:MMP-9可能参与骨肉瘤细胞s MICA蛋白形成,MMP-9可作为骨肉瘤免疫治疗的侯选靶点。

Objective To provide a new putative target for immunotherapy on osteosarcoma and explore the effect of matrix metalloproteinase-9 （MMP-9） on the formation of soluble MICA protein in osteosarcoma cells. Methods MMP-9 antisense oligonucleotide was transfected to osteosarcoma cells with lipofeetamine 2000. MMP-9 mRNA was assessed by RT-PCR. MMP-9 activity and soluble MICA （sMICA） in the culture supernatant was examined by zymography and quantified by ELISA, respectively. Results MMP-9 mRNA expression and gelatin enzymatic activity were inhibited in MMP-9 antisense oligonucleotide group. Comparing with the control group, lower concentration of sMICA was found in MMP-9 antisense oligonueleotide group （P = 0.011）. Conclusion MMP-9 may have an effect on the formation of sMICA protein in osteosarcoma. MMP-9 could be regarded as a putative target for immunotherapy in osteosarcoma.