CAJ | 학술논문

目的：探讨COX?2抑制剂Mavacoxib对骨肉瘤干细胞增殖、凋亡及相关信号通路的影响。方法体外培养人骨肉瘤MG?63细胞以获取骨肉瘤干细胞，经Mavacoxib（0、1、10、50、100μmol／L）处理后采用四甲基偶氮唑盐（MTT）比色法检测增殖抑制率的变化，同时采用流式细胞术Annexin?FITC／PI双染法及PI单染法检测不同浓度Mavacoxib处理24、48 h的凋亡情况（早、晚期凋亡率）及48 h的细胞周期情况， Western blotting检测不同浓度Mavacoxib 处理48 h的骨肉瘤干细胞中PI3K／Akt及Wnt／β?catenin信号通路的影响。结果在10～100μmol／L范围内，Mavacoxib可呈剂量和时间依赖的方式升高骨肉瘤干细胞的增殖抑制率，差异均有统计学意义（ P＜0?05）；与0μmol／L相比，除1μmol／L 24 h的晚期凋亡率无统计学差异，10～100μmol／L的早晚期凋亡率、G0／G1期细胞比例均升高，S期、G2／M期细胞比例均降低（ P＜0?05）。 Mavacoxib处理后的PI3K／Akt通路中的PTEN水平升高，Akt水平降低；Wnt／β?catenin通路中β?catenin、C?myc和 Cyclin D1水平均降低（ P＜0?05）。结论 Mavacoxib 对骨肉瘤干细胞有毒性作用，且可诱导其凋亡及细胞周期阻滞并抑制PI3K／Akt和Wnt／β?catenin信号通路的激活。
목적：탐토COX?2억제제Mavacoxib대골육류간세포증식、조망급상관신호통로적영향。방법체외배양인골육류MG?63세포이획취골육류간세포，경Mavacoxib（0、1、10、50、100μmol／L）처리후채용사갑기우담서염（MTT）비색법검측증식억제솔적변화，동시채용류식세포술Annexin?FITC／PI쌍염법급PI단염법검측불동농도Mavacoxib처리24、48 h적조망정황（조、만기조망솔）급48 h적세포주기정황， Western blotting검측불동농도Mavacoxib 처리48 h적골육류간세포중PI3K／Akt급Wnt／β?catenin신호통로적영향。결과재10～100μmol／L범위내，Mavacoxib가정제량화시간의뢰적방식승고골육류간세포적증식억제솔，차이균유통계학의의（ P＜0?05）；여0μmol／L상비，제1μmol／L 24 h적만기조망솔무통계학차이，10～100μmol／L적조만기조망솔、G0／G1기세포비례균승고，S기、G2／M기세포비례균강저（ P＜0?05）。 Mavacoxib처리후적PI3K／Akt통로중적PTEN수평승고，Akt수평강저；Wnt／β?catenin통로중β?catenin、C?myc화 Cyclin D1수평균강저（ P＜0?05）。결론 Mavacoxib 대골육류간세포유독성작용，차가유도기조망급세포주기조체병억제PI3K／Akt화Wnt／β?catenin신호통로적격활。
Objective To explore the effects of COX?2 inhibitor Mavacoxib on proliferation, apoptosis and related signaling pathways of human osteosarcoma stem cells. Methods The MG?63 cells were cultured in vitro to obtain osteosarcoma stem cells. The MTT assay was used to detect the proliferation inhibition rates after treatment with Mavacoxib(0, 1, 10, 50, 100 μmol/L). Mean?while, Annexin?FITC/PI double staining and PI staining were used to detect the apoptotic rates at 24, 48 h and cell cycle at 48 h after different concentrations of Mavacoxib. The effects of Mavacoxib on the Wnt/β?catenin and PI3K/Akt signaling pathway were measured by Western blotting. Results In the range of 10?100μmol/L, Mavacoxib could improve the proliferation inhibition rates of osteosarco?ma stem cells in a dose and time dependent manner with statistical significant difference( P<0?05) . Compared with 0μmol/L, the ap?optotic rates and the percentage of G0/G1 phase cells were increased, the proportions of S phase and G2/M phase were decreased in other concentrations( P<0?05) . After the treatment of Mavacoxib, the levels of PTEN in the PI3K/Akt pathway were increased, the levels of Akt in the PI3K/Akt pathway and the levels of β?catenin, C?myc and Cyclin D1 in the Wnt/β?catenin pathway were de?creased( P<0?05) . Conclusion Mavacoxib has toxic effect on human osteosarcoma stem cells, and can induce apoptosis and cell cy?cle arrest and inhibit the activation of Wnt/β?catenin and PI3K/Akt signaling pathway.