摘要目的 观察5-脂氧合酶(5-lipoxygenase,5-LOX)抑制剂去甲二氢愈创木酸(nordihydroguaiaretic acid,NDGA)联合选择性环氧合酶-2(cyclooxygenase-2,COX-2)抑制剂塞来昔布(Celecoxib)对结肠癌细胞株HT-29细胞凋亡的影响.方法 用不同浓度NDGA及塞来昔布处理HT-29细胞,应用甲基偶氮唑蓝(MTT)法、倒置相差显微镜观察、Annexin V/PI法和RT-PCR方法,研究其抑制HT-29细胞增殖,诱导凋亡的作用.结果 MTT法显示,与对照组相比,NDGA和塞来昔布及两药联用组细胞活性下降(0.432±0.024、0.425±0.013、0.303±0.014 vs 0.693±0.018,t＝18.79,25.75,37.64,P＜0.01).NDGA联合塞来昔布组与单独应用NDGA或塞来昔布差异有统计学意义(t＝10.21,14.14,P＜0.01).倒置相差显微镜下显示两种药物都能使体外培养的结肠癌HT-29细胞的形态发生明显变化,而两种药物联合应用后变化更明显.激光共聚焦显微镜下显示应用NDGA及塞来昔布后可以引起HT-29细胞凋亡.RT-PCR检测发现药物作用后Caspase-3表达上调,两种药物联合应用后上调最多.结论 MDGA联合塞来昔布较之单独应用可以更强抑制体外培养的HT-29细胞增殖,并诱导凋亡,其抑制HT-29细胞诱导凋亡机制与激活Caspase-3途径有关.

abstractsObjective To study the effect of 5-lipoxygenase(5-LOX) inhibitor nordihyroguaiaretic acid (NDGA) combined the selective cyclooxygenase-2 (COX-2) inhibitor Celecoxib on the apoptosis of human colon carcinoma cell line HT-29. Methods Different concentration of NDGA and Celecoxib combinations were used to process cancer cell, and thiazolyl blue tetrazlium bromide (MTT) and phase contrast microscope and Annexin V/PI fluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to study the proliferation inhibited effect and apoptosis induced effect caused by combination of NDGA combined Celecoxib. Results MTT results showed that the viability of NDGA group, Celecoxib group and the group of NDGA combined Celecoxib (0.432±0.024,0.425±0.013,0.303±0.014 vs 0.693±0.018,t＝18.79,25.75,37.64,P＜0.01) was obviously lower than control group. The group of NDGA combined Celecoxib was significantly lower than NDGA group or Celecoxib group (t＝10.21, 14.14,P＜0.01). Under inverted phase contrast microscope, cell morphology significantly changed, and the group of NDGA combined Celecoxib changed most obviously. Apoptosis was observed by laser scanning confocal microscope (LSM) after NDGA and Celecoxib were used to process the HT-29. RT-PCR showed that up-regulation of Caspase-3 after treatment, and the combination of two drugs increased the most. Conclusions NDGA combined Celecoxib inhibited proliferation and induced apoptosis in human colon carcinoma cell line HT-29, and combined therapy had better effect than that of any drug used separate-ly. The mechanism may be associated with up-regulation of Caspase-3.