摘要
目的探讨不同温度热疗对宫颈癌Caski细胞坏死和凋亡的影响。方法采用水浴对宫颈癌Caski细胞行43℃,45℃,47℃实验组和37℃对照组热疗,四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)检测细胞增殖抑制率,流式细胞仪测定细胞凋亡和坏死率,免疫组化检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达。结果45℃,47℃组的细胞增殖抑制率在24h、48h、72h、96h分别为76.11%±5.32%、81.08%±4.03%、74.71%±3.78%、72.16%±4.75%和88.57%±3.77%、91.33%±2.43%、91.09%±1.17%、92.05%±1.67%,明显高于43℃组的26.57%±3.46%、25.49%±2.76%、22.80%±3.16%、26.87%±4.01%(q=10.41,18.23,14.53,13.57,P<0.05;q=14.41,21.62,16.03,16.01,P<0.05)。45℃与47℃组细胞增殖抑制率在24h、72h、96h的差异无显著性(q=3.01,1.49,2.44,P>0.05)。45℃组的细胞凋亡率最高,为12.82%±1.77%,与其他各组之间的差异具有显著性(F=18.69,P<0.05)。47℃组的细胞坏死率最高,为39.15%±5.67%,各组之间的差异均有显著性(F=69.16,P<0.05)。PCNA的表达随着温度的增加而降低,45℃,47℃组的平均光密度值(average optical density,AOD)分别是0.22±0.02、0.21±0.01,与37℃对照组0.28±0.02相比差异具有显著性(q=4.75,5.76,P<0.05)。结论45℃及以上的热疗能够明显抑制Caski细胞的增殖,增加凋亡和坏死率,降低PCNA的表达。
Objective To investigate the effect of hyperthermia at different temperatures on the proliferation and apoptosis of Caski cells. Methods Caski cells were incubated at 43 ℃, 45 ℃ , 47℃ , or 37 ℃ (control) in temperature-controlled water baths. The cell growth, apoptosis rate, and expression of PCNA protein were measured by MTT, flow cytometry, and immunohistochemistry respectively after the therapy. Results The inhibition rates of the cell growth in 45℃ and 47℃ groups at hours 24, 48, 72, and 96 were 76.11%±5.32%, 81.08%±4.03%, 74.71%±3.78%, and 72. 16%±4.75%, and 88.57%± 3.77% , 91.33%±2.43% , 91.09%± 1.17% , and 92. 05%± 1.67% , respectively, which were significantly higher than those in the 43℃ group 26.57%±3.46%, 25.49%±2.76%, 22.80%±3.16%, and 26.87%±4.01% (q = 10.41, 18.23, 14.53, and 13.57, P 〈 0.05 ; and q = 14.41, 21.62, 16.03, and 16.01, P 〈 0.05 ; respectively). The difference in the inhibition effect of cell growth between 45℃ and 47℃ groups was not significant at 24, 72, and 96 hours (q =3.01, 1.49, 2.44, P 〉0.05). The apoptosis rate in the 45℃ group (12.82% ± 1.77% ) was significantly higher than the other groups (F = 18.69, P 〈0.05) ; while the highest necrosis rate was found in the 47℃ group (39.15% ± 5.67%, F =69.16, P 〈 0.05). The expression of PCNA decreased with the temperature of the bath. The average optical density of PCNA was 0. 22 ± 0.02 and 0.21 ± 0. 01 respectively in the 45℃ and 47℃ groups, which were significantly higher than that in the control (0.28 ±0.02, q =4.75, 5.76, P 〈0.05). Conclusions Hyperthermia at 45 ℃ or higher temperature can remarkably inhibit the proliferation of Caski cells, increase the rates of apoptosis and necrosis, and decrease the expression of PCNA protein.
出处
《中国微创外科杂志》
CSCD
2009年第3期254-257,共4页
Chinese Journal of Minimally Invasive Surgery
基金
国家自然科学基金(10775085)
国家自然科学基金(30571799)
北京市科委(Z07000200540704)
清华大学裕元基金