塞来昔布对鼻咽癌CNE-2Z细胞生物学行为的影响

目的 探讨塞来昔布对人鼻咽癌CNE-2Z细胞增殖、黏附、运动能力等生物学行为的影响.方法 不同浓度塞来昔布作用鼻咽癌细胞24 h后,运用平板克隆形成实验检测塞来昔布对鼻咽癌CNE-2Z细胞增殖的影响;应用细胞-细胞黏附实验、细胞-基质黏附实验和细胞运动实验检测塞来昔布对肿瘤细胞黏附和运动能力的影响;Western blot和免疫细胞化学检测鼻咽癌细胞COX-2蛋白表达变化.结果 塞来昔布可以明显抑制鼻咽癌CNE-2Z细胞的增殖,0、25、50、75、100 μmol/L组肿瘤细胞克隆形成数率分别为(85.68±1.83)%、(69.00±4.08)%、(59.03±2.43)%、(37.28±3.25)%和(12.58±3.69)%,各组间比较差异均有显著性(F=321.187,P＜0.01).细胞-细胞黏附实验结果显示,同一个时间点各组间细胞黏附率差异有显著性(F值分别为21.605、181.301,P均＜0.001),其中15 min时间点0、25、50 μmol/L组细胞间黏附率分别为(12.38±1.54)%、(19.30±1.49)%和(20.18±1.80)%,45 min时间点细胞间黏附率分别为(46.34±1.37)%、(58.72±1.68)%和(67.15±0.86)%;细胞-基质黏附实验结果表明,25、50 μmol/L组细胞异质黏附力明显低于无药组(t值分别为24.840和38.565,P均＜0.01);运动实验显示穿膜肿瘤细胞数分别为(124.3±8.02)个、(84.3±12.22)个和(55.3±9.29)个,组间差异有统计学意义,用药组穿膜细胞数明显减少(F＝36.010,P＜0.01);Western blot结果发现,25、50 μmol/L组COX-2蛋白的表达水平与对照组相比明显下降(t值分别为25.356和73.656,P均＜0.01);免疫细胞组织化学结果发现,0、25、50 μmol/L组COX-2阳性表达数分别为(380±11)个、(313±12)个、(213±8)个,各浓度组间差异有统计学意义(F＝298.578,P＜0.01).结论 塞来昔布可以抑制鼻咽癌细胞的增殖,增强鼻咽癌细胞间的黏附力,降低细胞与细胞外基质的黏附力及运动能力,机制可能与下调COX-2蛋白的表达水平有关.

Effects of celecoxib on the biologic behavior of nasopharyngeal carcinoma cell CNE-2Z

Purpose To evaluate the effect of celecoxib(a selective cyclooxygenase-2 inhibitor) on the proliferation,adhesion and invasion of human nasopharyngeal carcinoma(NPC) cell line CNE-2Z.Methods After treated with different concentrations of celecoxib for 24 hours,we used the colony formation assay to detect the effect of celecoxib on CNE-2Z cell proliferation,and the cell-cell adhesion assay,cell-matrix adhesion assay and migration assay to investigate the effects of celecoxib on adhesive and invasive capacity of CNE-2Z cell.Western blot and immunocytochemistry were used to detect the expression levels of COX-2.Results The colony formation assay showed that celecoxib could greatly inhibit CNE-2Z cell proliferation,the rate of colony formation was(85.68±1.83)%,(69.00±4.08)%,(59.03±2.43)%,(37.28±3.25)% and(12.58±3.69)% in 0,25,50,75,and 100 μmol/L groups,respectively(F=321.187,P<0.01).The cell-cell adhesion assay indicated that there was a significant difference between the experimental and control groups at the same time(F=321.187,P<0.01).At 15 min time point,the cell adhesion rate was(12.38±1.54)%,(19.30±1.49)% and(20.18±1.80)% in 0,25,50 μmol/L group,respectively.At 45 min time point,there was(46.34±1.37)%,(58.72±1.68)% and(67.15±0.86)% in 0,25,50 μmol/L group,respectively.The cell-matrix adhesion assay showed that the cell-matrix adhesion ability was significantly inhibited in 25,50 μmol/L group compared to the control group(t=24.840,38.565,respectively,P<0.01).The migration assay showed that there was a significant difference between the experimental and control groups(F=36.010,P<0.01),the cell numbers passing through PVPF were(124.3±8.02),(84.3±12.22) and(55.3±9.29) in 0,25,50 μmol/L group,respectively.Western blot showed that COX-2 protein was significantly downregulated in 25,50 μmol/L group(t=25.356,73.656,respectively,P<0.01).Immunocytochemistry results showed that the expression levels of COX-2 were(380±11),(313±12)and(213±8) in 0,25,and 50 μmol/L groups,respectively,indicating a significant difference between groups(F=298.578,P<0.01).Conclusions Celecoxib can improve the cell-cell adhesion capacity,and inhibit the cell-matrix adhesion and migration capacity of CNE-2Z cell,which might be due to the inhibition of COX-2 expression.