人骨肉瘤MG-63细胞自分泌可溶性纤连蛋白参与细胞形态维持

Roles of autocrine soluble fibronectin on the maintenance of cell shape of osteosarcoma MG-63 cells

目的:探讨MG-63细胞自分泌可溶性纤连蛋白(FN)在细胞形态维持中的作用。方法:将培养24 h MG-63细胞随机分成对照组、更换新鲜培养基组(不含自分泌可溶性FN)、更换培养MG-63细胞24 h的条件培养基组(含自分泌可溶性FN)和更换添加FN的新鲜培养基组(添加FN),在0.5 h、1 h和2 h观察细胞形态;采用ELISA法检测新鲜和条件培养基中可溶性FN含量;采用免疫荧光方法观察细胞微丝骨架和不溶性FN的分布变化。结果:检测新鲜和条件培养基可溶性FN浓度结果表明,条件培养基中可溶性FN含量明显高于新鲜培养基(P<0.01);与对照组相比,不含自分泌可溶性FN组细胞0.5 h时回缩明显,由梭形变成圆形,1 h时开始重新铺展,2 h时形态恢复明显;含自分泌可溶性FN组和添加FN组细胞形态没有明显变化;观察细胞微丝骨架和不溶性FN分布发现,与对照组相比,不含自分泌可溶性FN组细胞0.5 h时应力纤维解聚,细胞外纤丝状FN明显减少。结论:细胞自分泌可溶性FN可能通过参与调节不溶性FN和微丝骨架的重构,参与细胞形态调节。

AIM： To investigate the roles of autocrine soluble FN of MG-63 cells in cell shape maintaining.METHODS： Human osteosarcoma MG-63 cells（1×104 cells/cm2） were routinely cultured in MEM medium＋10% fetal bovine serum（FBS） for 24 h and then the medium was replaced by fresh medium（the group without autocrine soluble FN）,conditional medium of collected after 24 h of MG-63 cells（the group containing autocrine soluble FN） and fresh medium with 20 μg/mL FN（the group with plasma FN） after 24 h,and the cell morphology was observed using phase contrast microscope;The concentrations of soluble FN in fresh and conditional medium were detected;Microfilament and insoluble FN changes of MG-63 cells were investigated.RESULTS： The concentration of soluble FN in conditional medium is much higher than that in the fresh medium（P0.01）.Fresh medium will result in the cell shape changing from spindle to round for 0.5 h.The cell began to spread for 1 h,and the cell shape recovered for 2 h;Conditioned medium has no significant effects on cell shape.Addition of FN（20 mg/L） to fresh medium could inhibit the cell shape change induced by fresh medium.Microfilament and insoluble FN were disorganized by fresh medium.CONCLUSION： These results indicated that autocrine soluble FN of osteosarcoma MG-63 cells may be involved in cell shape maintaining.