三氧化二砷联合COX-2抑制剂塞来昔布对高转移卵巢癌细胞系HO-8910PM的体内、体外抗肿瘤实验研究

In vitro and in vivo anti-neoplasm effects of arsenic trioxide combined with COX-2 inhibitor on highly metastatic ovarian cancer cell line HO-8910PM

目的探索三氧化二砷和COX-2抑制剂塞来昔布对高转移性卵巢癌细胞的联合抗肿瘤效应。方法在体外实验中,三氧化二砷和塞来昔布作用于HO-8910PM细胞后,采用噻唑兰(MTT)比色实验、Hoechest33258凋亡染色实验、流式细胞术等方法在细胞水平检验这两种药物单药及联合的增殖抑制和诱导凋亡作用。体内实验中,通过腋窝皮下注射HO-8910PM细胞建立BALB/c-nu裸鼠移植瘤模型,并随机分为阴性对照组、AS2O3单药组、Celecoxib单药组以及AS2O3联合Celecoxib组等四个处理组,AS2O3和Celecoxib分别采用腹腔灌注和灌胃的方法,实验结束时称取各组移植瘤的瘤重,计算抑瘤率并进行组间统计学比较分析。结果三氧化二砷和塞来昔布对HO-8910PM细胞株均具有剂量依赖性的增殖抑制作用,IC50值分别为65.48μmol/L和49.13μmol/L。两药联合后的增殖抑制率均大于任一单药抑制率且差异具有统计学意义(P<0.05),而联合用药的q值分别为3.23和2.42,均大于1.15,提示两药联合具有显著的协同抗肿瘤效应。Hoechest33258凋亡染色和流式细胞仪检测从凋亡角度进一步证实了MTT的实验结果。裸鼠实验中,联合用药组相对于单药组的抗肿瘤效应未显示出统计学差异,但联合用药组的平均瘤重仍为最小。结论三氧化二砷联合塞来昔布对高转移性卵巢癌细胞HO-8910PM具有明确的协同抗肿瘤作用,该用药方案对高转移性的卵巢癌具有进一步的研究价值。

Objective To investigate the anti-neoplasm effects of the combination of arsenic trioxide and COX-2 inhibitor（celecoxib） on highly metastatic ovarian cancer cell line HO-8910 PM. Methods In the in vitro test,the HO-8910 PM cells were treated with AS2O3 or celecoxib or both agents. Cell proliferation was assessed by MTT assay. Cell apoptosis was detected by flow cytometry and Hoechest33258 fluorescence staining. In the in vivo test, the nude mice bearing transplanted tumors were divided into control group, AS2O3 group, celecoxib group, and AS2O3 plus celecoxib group, and were intraperitoneally injected with AS2O3and（or） intragastrically administered with celecoxib.The weight of xenografts was measured after treatment and the inhibition rate was calculated. Results Dose-dependent anti-proliferative effects were observed in MTT test. The IC50 value of AS2O3 was 65.48 μmol/L, and that of celecoxib was 49.13 μmol/L. When celecoxib and AS2O3 were applied simultaneously, the proliferative inhibition rate was higher than that of the group treated with celecoxib or AS2O3alone（P〈0.05）, and the q values were 3.23 and 2.42, respectively, both of which were greater than 1.15. The apoptotic results of flow cytometry and Hoechest33258 fluorescence staining proved the prior results from MTT test. In the in vivo test, there were no significant differences between single-agent group and two-agent group, but the average tumor weight of the two-agent group was also the minimum in the four groups. Conclusion There are synergistic effects when arsenic trioxide and celecoxib are used in combination in highly metastatic ovarian cancer cell line HO-8910 PM. The combined regimen of the two agents is worth further research.