南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (06): 792-.

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塞来昔布拮抗卡铂在食管癌细胞的细胞毒作用

时粒笠,钟德胜,古春萍,余乐   

  • 出版日期:2014-06-20 发布日期:2014-06-20

Celecoxib antagonizes the cytotoxic effect of carboplatin in human esophageal cancer
cells

  • Online:2014-06-20 Published:2014-06-20

摘要: 目的探讨塞来昔布(celecoxib)在人食管癌细胞是否影响卡铂介导的细胞毒作用。方法MTT法检测卡铂单独及联合
celecoxib 后对食管癌亲代细胞EC109 及耐药细胞EC109/CDDP 的细胞毒作用。Western blot 方法检测卡铂单独及联合
celecoxib 后caspase-3 的活化、PARP蛋白的剪切,以及CTR1 蛋白的表达。Caspase-3/7 Assay 试剂盒检测细胞caspase-3 酶活
性。流式细胞仪检测卡铂单独及联合celecoxib作用下细胞凋亡比率。ICP-MS检测卡铂单独及联合celecoxib作用下细胞内卡
铂蓄积量。结果卡铂在EC109及EC109/CDDP细胞的IC50在联合应用celecoxib后显著增加,说明celecoxib拮抗了卡铂介导
的细胞毒作用。在联合应用celecoxib后caspase-3及PARP蛋白的剪切较单独应用卡铂时明显减少,且caspase-3酶活性较单独
应用卡铂时显著降低。此外,联合应用celecoxib后,EC109和EC109/CDDP细胞凋亡率比单独应用卡铂时明显降低,而细胞存
活率明显增加。联合应用celecoxib后,EC109和EC109/CDDP细胞内卡铂蓄积量比单独应用卡铂时显著降低,而CTR1蛋白的
表达无明显改变。结论Celecoxib拮抗卡铂在食管癌细胞介导的细胞毒作用,通过降低细胞内卡铂蓄积量抑制卡铂介导的食
管癌细胞凋亡。

Abstract: Objective To explore the antagonizing effect of celecoxib against the cytotoxicity of carboplatin in human
esophageal cancer cells. Methods The cell viability of cisplatin-resistant cell line EC109/CDDP and its parental cell line EC109
exposed to carboplatin alone or carboplatin plus celecoxib was determined by MTT assay. The expression of CTR1, caspase-3
activation and PARP cleavage in the exposed cells were examined by Western blotting. Caspase-3 activity and cell apoptosis
after the exposure were detected with Caspase-3/7 assay and flow cytometry, respectively. The effect of celecoxib on
carboplatin accumulation in the cells was measured using inductively coupled plasma mass spectrometry (ICP-MS). Results
Celecoxib treatment significantly increased the IC50 of carboplatin, suppressed carboplatin-induced caspase-3 and PARP
cleavage and caspase-3 activity in EC109 and EC109/CDDP cells. Celecoxib also inhibited carboplatin-induced apoptosis and
suppressed intracellular carboplatin accumulation in both cell lines. A combined exposure to celecoxib and carboplatin did not
cause significant changes in the protein expression of CTR1. Conclusion Celecoxib antagonizes the cytotoxic effect of
carboplatin and inhibits carboplatin-induced apoptosis in human esophageal cancer cells by reducing intracellular carboplatin
accumulation.