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目的 :检测二萜类化合物冬凌草甲素联合环氧合酶-2(cyclooxygenase-2,Cox-2)抑制剂塞来昔布对人骨肉瘤MG-63细胞增殖及凋亡的影响,并探讨可能的作用机制。方法 :MTT法检测不同浓度的冬凌草甲素(20、40和60μmol/L)和塞来昔布(25、50和100μmol/L)单药和联合处理12和24 h后MG-63细胞的增殖抑制率;FCM法检测冬凌草甲素(40μmol/L)和塞来昔布(50μmol/L)单药和联合干预MG-63细胞后的凋亡率,并分别采用实时荧光定量PCR法和蛋白质印迹法检测对Cox-2以及凋亡调节因子[B细胞性淋巴瘤-2(B cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)和生存素(survivin)]m RNA及蛋白表达水平的影响。结果 :不同浓度的冬凌草甲素和塞来昔布单药和联合干预后,MG-63细胞的增殖明显受到抑制(P值均<0.05),呈剂量依赖性,且在低剂量和中剂量时两药有协同作用。冬凌草甲素和塞来昔布联合用药组较空白对照组和各单药组细胞的凋亡率增加(P值均<0.05)。联合用药组中Cox-2、Bcl-2和survivin m RNA及蛋白的表达水平均较空白对照组和各单药组明显下调(P值均<0.05),而Bax m RNA及蛋白的表达水平明显上调(P值均<0.05)。结论 :冬凌草甲素联合塞来昔布可协同抑制MG-63细胞,这一过程可能与Cox-2的表达受到抑制和凋亡调节因子表达水平的改变有关。
Objective: To investigate the effect of diterpenoid Oridonin and cyclooxygenase-2 (Cox-2) inhibitor celecoxib on the proliferation and apoptosis of human osteosarcoma MG-63 cells Possible mechanism of action. METHODS: MG-63 cells were treated with different concentrations of oridon (20, 40 and 60 μmol / L) and celecoxib (25, 50 and 100 μmol / L) . The apoptosis rate of MG-63 cells treated with oridon (40μmol / L) and celecoxib (50μmol / L) alone or in combination with FCM was detected by real-time fluorescence quantitative PCR Western blotting was used to detect the expression of Cox-2 and Bcl-2 associated protein B (Bcl-2, Bcl-2, Bcl- ) And survivin] m RNA and protein expression levels. Results: The proliferation of MG-63 cells was significantly inhibited by different concentrations of oridonin and celecoxib alone (all P <0.05), in a dose-dependent manner. At low dose and medium dose The two drugs have a synergistic effect at the dose. Oridonin and celecoxib combination treatment group compared with the blank control group and the single drug group increased the rate of apoptosis (P values were <0.05). The expression levels of Cox-2, Bcl-2 and survivin m RNA and protein in combination group were significantly lower than those in blank control group and single drug group (P <0.05), while the expression of Bax m RNA and protein were significantly (All P <0.05). CONCLUSION: Oridonin combined with celecoxib can inhibit the proliferation of MG-63 cells in a dose-dependent manner. This process may be related to the inhibition of Cox-2 expression and the change of expression of apoptosis-regulatory factors.