选择性及非选择性COX-2抑制剂对结肠癌细胞生长的影响

Effects of Selective and Non-selective Cyclooxygenase-2 Inhibitor on the Growth of Colon Cancer Cells

目的探讨选择性及非选择性环氧合酶-2(cy looxygenase-2,COX-2)抑制剂对体外培养的结肠癌细胞生长的影响及凋亡的诱导作用。方法体外培养HT-29、SW 480及LS174-T三种结肠癌细胞,分别加入不同浓度的非选择性COX-2抑制剂阿司匹林(asp irin)、选择性COX-2抑制剂塞来昔布(ce lecox ib)及美洛昔康(m e lox icam)作用于HT-29及SW 480细胞,采用M TT法检测结肠癌细胞增殖;用免疫细胞化学法及免疫印迹技术分别检测结肠癌细胞增殖细胞核抗原(PCNA)及细胞COX-2的表达;流式细胞技术分析对结肠癌细胞周期的影响;用A nnex inV/P I染色结合流式细胞术检测细胞凋亡。结果asp irin、ce lecox ib及m e lox icam均能有效抑制体外培养的HT-29、SW 480结肠癌细胞生长,并具有良好的量-效关系。W estern b lot表明,HT-29细胞表达COX-2,而SW 480细胞不表达COX-2。A sp irin及ce lecox ib处理组细胞PCNA表达较对照组明显下调。10 mm o l/L的asp irin及50μm o l/L的ce lecox ib能诱导HT-29及SW 480细胞凋亡,凋亡率分别为4.8%,17.7%;5.1%,20.4%。结论选择性及非选择性COX-2抑制剂均能有效抑制结肠癌细胞生长,这种作用亦存在于COX-2阴性表达的结肠癌细胞。

Objective To evaluate the effects of selective and non-selective cyclooxygenase-2 on the growth and apoptosis of colon cancer cell lines in vitro. Methods The proliferation of colon cancer cells was determined by MTT assay, and the cell cycle progression was analyzed by flow cytometric assay. Annexin V/PI staining in combination with flow cytometric assay was used to detect apoptosis induced by NSAIDs. Results It was found that celecoxib, meloxicam and aspirin could inhibit the growth of HT-29 or SW480 cell and showed a concentration dependent pattern. COX-2 protein was expressed in HT-29 and LS174-T, but not in SW480 cells. In addition, PCNA levels in both HT-29 cell and SW480 cells were reduced by aspirin and celecoxib. Both aspirin （10 mmol/L） and celecoxib （50 μmol/L） induced apoptosis of HT-29 and SW480 colon cancer cells, the apoptosis rates were 4.8%, 17.7% and 5.1%, 20.4% respectively. Conclusion Both COX-2 selective and non-selective inhibitors can potentially inhibit the growth of colon cell lines and such inhibitory effect on COX-2 negative colon cancer cells is also evident.