CAJ | 학술논문

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。
목적：탐토새래석포（celecoxib）재인식관암세포시부영향잡박개도적세포독작용。방법 MTT법검측잡박단독급연합celecoxib후대식관암친대세포EC109급내약세포EC109/CDDP적세포독작용。Western blot방법검측잡박단독급연합celecoxib후caspase-3적활화、PARP단백적전절，이급CTR1단백적표체。Caspase-3/7 Assay시제합검측세포caspase-3매활성。류식세포의검측잡박단독급연합celecoxib작용하세포조망비솔。ICP-MS검측잡박단독급연합celecoxib작용하세포내잡박축적량。결과잡박재EC109급EC109/CDDP세포적IC50재연합응용celecoxib후현저증가，설명celecoxib길항료잡박개도적세포독작용。재연합응용celecoxib후caspase-3급PARP단백적전절교단독응용잡박시명현감소，차caspase-3매활성교단독응용잡박시현저강저。차외，연합응용celecoxib후，EC109화EC109/CDDP세포조망솔비단독응용잡박시명현강저，이세포존활솔명현증가。연합응용celecoxib후，EC109화EC109/CDDP세포내잡박축적량비단독응용잡박시현저강저，이CTR1단백적표체무명현개변。결론 Celecoxib길항잡박재식관암세포개도적세포독작용，통과강저세포내잡박축적량억제잡박개도적식관암세포조망。
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.