CAJ | 학술논문

目的探讨阻断转化生长因子-β(TGF-β)信号通路对胶质瘤血管生成拟态(VM)的影响及其可能机制. 方法三维培养人脑胶质瘤细胞系U251、SHG44,观察U251培养上清、TGF-β因子对SHG44细胞形成VM的影响；比较0μg/mL(PBS组)、15 μg/mL(Ab15组)、30 μg/mL (Ab30组)TGF-β中和抗体对U251、SHG44细胞形成VM的影响；酶联免疫吸附法(ELISA)检测空白组、PBS组、Ab15组,Ab30组U251培养上清中血管内皮生长因子(VEGF)及血小板衍生生长因子(PDGF)的表达以及空白组、TGF-β组、PBS组、Ab15组,Ab30组SHG44细胞上清VEGF、PDGF的表达. 结果三维培养时,U251细胞排列形成VM,SHG44细胞并不形成VM,而仅仅相互聚集成大小不等的细胞集落；U251培养上清可以诱导SHG44细胞形成VM,且在培养24～48 h最为明显,TGF-β因子不能诱导SHG44细胞形成VM;PBS组、Ab15组、Ab30组U251细胞环状结构数量依次降低,差异有统计学意义(P＜0.05).与U251上清共培养的SHG44细胞在加入TGF-β中和抗体后,3组细胞形成环状结构数量也依次降低,差异有统计学意义(P＜0.05)；与空白组和PBS组比较,Ab15组、Ab30组U251细胞上清中VEGF、PDGF浓度下降,差异有统计学意义(P＜0.05),与空白组和TGF-β组相比,PBS组、Ab15组、Ab30组SHG44细胞上清中两种因子浓度均增高,差异有统计学意义(P＜0.05). 结论抑制TGF-β信号通路可以使胶质瘤VM形成能力下降,这可能与VEGF及PDGF表达减少有关.
목적 탐토조단전화생장인자-β(TGF-β)신호통로대효질류혈관생성의태(VM)적영향급기가능궤제. 방법 삼유배양인뇌효질류세포계U251、SHG44,관찰U251배양상청、TGF-β인자대SHG44세포형성VM적영향；비교0μg/mL(PBS조)、15 μg/mL(Ab15조)、30 μg/mL (Ab30조)TGF-β중화항체대U251、SHG44세포형성VM적영향；매련면역흡부법(ELISA)검측공백조、PBS조、Ab15조,Ab30조U251배양상청중혈관내피생장인자(VEGF)급혈소판연생생장인자(PDGF)적표체이급공백조、TGF-β조、PBS조、Ab15조,Ab30조SHG44세포상청VEGF、PDGF적표체. 결과 삼유배양시,U251세포배렬형성VM,SHG44세포병불형성VM,이부부상호취집성대소불등적세포집락；U251배양상청가이유도SHG44세포형성VM,차재배양24～48 h최위명현,TGF-β인자불능유도SHG44세포형성VM;PBS조、Ab15조、Ab30조U251세포배상결구수량의차강저,차이유통계학의의(P＜0.05).여U251상청공배양적SHG44세포재가입TGF-β중화항체후,3조세포형성배상결구수량야의차강저,차이유통계학의의(P＜0.05)；여공백조화PBS조비교,Ab15조、Ab30조U251세포상청중VEGF、PDGF농도하강,차이유통계학의의(P＜0.05),여공백조화TGF-β조상비,PBS조、Ab15조、Ab30조SHG44세포상청중량충인자농도균증고,차이유통계학의의(P＜0.05). 결론 억제TGF-β신호통로가이사효질류VM형성능력하강,저가능여VEGF급PDGF표체감소유관.
Objective To study the effect of transforming growth factor-β (TGF-β) signaling pathway blockage on vasculogenic mimicry (VM) in gliomas and explore its possible mechanism.Methods Three-dimentional culture was performed on the glioma cell lines U251 and SHG44; the effects of U251 culture supematant and TGF-β on VM formation of SHG44 cells were observed; the capability of VM formation of U251 and SHG44 cells after being treated with 0 μg/mL (PBS group),15 μg/mL TGF-β neutralizing antibody (Ab15 group) and 30 μg/mL TGF-β neutralizing antibody (Ab30 group) was evaluated.ELISA was used to detect the concentrations of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) in the supematant of U251 cells from the blank group,PBS group,Ab 15 group and Ab30 group and the concentrations of VEGF and PDGF in the supernatant of SHG44 cells from the blank group,TGF-β treatment group,PBS group,Ab15 group and Ab30 group.Results VM was formed in the U251 cells while not in the SHG44 cells during the three-dimentional culture; SHG44 cells could only gather into colonies of different sizes.U251 culture supernatant could induce SHG44 cells to form VM,enjoying the most obvious effect at 24-48 h of culture; TGF-β could not induce SHG44 cells to form VM.The number of U251 cells annulation in PBS group,Ab15 group and Ab30 group decreased in sequence with significant difference (P＜0.05).The number of U251 cells armulation in SHG44 cells cultured in U251 culture supematant from the PBS group,Ab15 group and Ab30 group decreased in sequence after being added TGF-β antibody with significant difference (P＜0.05).As compared with that in the blank group and PBS group,significant decrease of VEGF and PDGF concentrations in the U251 cells from Ab15 group and Ab30 group was noted (P＜0.05); as compared with that in the blank group and TGF-β treatment group,significant increase of VEGF and PDGF concentrations in the SHG44 cells from PBS group,Ab15 group and Ab30 group was noted.Conclusion Blockage of TGF-β signaling pathways inhibits VM in glioma,and it maybe probably due to the decrease of VEGF and PDGF expressions..