重组人血管抑素抑制人鼻咽癌生长的实验研究

目的：观察重组人血管抑素（ｒｅｃｏｍｂｉｎａｎｔ ｈｕｍａｎ ａｎｇｉｏｓｔａｔｉｎ,ｒｈＡＧＮ）对鼻咽癌皮下移植瘤生长的抑制作用,探讨其应用价值。方法：应用ＭＴＴ法观察ｒｈＡＧＮ对ＣＮＥ－１鼻咽癌细胞和ＨＤＭＥＣ血管内皮细胞增殖的影响;建立ＣＮＥ－１鼻咽癌裸鼠皮下移植瘤模型,皮下注射不同剂量ｒｈＡＧＮ,治疗期为３０天,观察肿瘤体积和重量变化。结果：（１）在０．５μｇ／ｍｌ、１．０μｇ／ｍｌ和２．     

土贝母苷甲诱导人鼻咽癌细胞CNE-2Z凋亡

背景和目的：土贝母[Bolbostemma paniculatum(Maxim．)Franquet]是一种传统中药,我们以前的研究结果已证实从中药土贝母块茎中分离得到的土贝母苷甲(下称苷甲)有强抗肿瘤效果。本研究旨在探讨苷甲对人鼻咽癌细胞株CNE-2Z细胞凋亡的影响。方法：MTT法检测苷甲对肿瘤细胞生长的抑制作用,流式细胞仪分析细胞周期,荧光显微镜和电子显微镜观察细胞形态,琼脂糖凝胶电泳检测DNA电泳图谱的变化,蛋白免疫印迹法检测凋亡相关基因bcl-2和bax表达的变化。结果：苷甲抑制CNE-2Z细胞的生长,其24、48、72h的IC50分别为32．5、20．7、16.7μmol／L。苷甲诱导CNE-2Z细胞发生程序性死亡：流式细胞分析发现Sub—G1峰,50μmol／L苷甲作用CNE-2Z细胞12h,凋亡指数为72．8％;CNE-2Z细胞经苷甲作用(10μmol／L,24、48、72h;30、40、50、60μmol／L,12h)后,其DNA电泳图中出现典型的梯形条带。苷甲诱导bcl-2表达下调且磷酸化,并伴有bax高表达。结论：苷甲诱导CNE-2Z细胞发生形态学和生化学上典型的程序性死亡,其诱导的CNE-2Z细胞凋亡与bcl-2失活和bax激活有关。     

沙利度胺对人鼻咽癌细胞株血管内皮生长因子表达影响的观察

目的:探讨沙利度胺对人鼻咽癌细胞株体外生长及对血管内皮生长因子表达的影响。方法:采用MTT法检测沙利度胺对人鼻咽癌细胞增殖的抑制率;RT-PCR和免疫细胞化学方法分别检测沙利度胺作用前后血管内皮生长因子基因和蛋白表达水平的变化。结果:沙利度胺能抑制CNE-2细胞增殖,呈浓度依赖性,当60mg/L沙利度胺作用72h,其抑制率为45.6%;不同浓度沙利度胺作用后VEGFmRNA和VEGF-CmRNA表达均降低,VEGFmRNA下降最明显,P<0.05;蛋白表达与其相应mRNA呈正相关,r值分别为0.959和0.974,P值分别为0.035和0.021。结论:沙利度胺能够抑制CNE-2细胞增殖,其作用机制可能与其下调血管内皮生长因子的表达有关。     

姜黄素对人鼻咽癌CNE-2Z细胞侵袭和转移的体外实验研究

背景与目的:鼻咽癌放射治疗后的局部复发和远处转移是患者死亡的主要原因之一。姜黄素(curcumin)是从姜科姜黄属植物姜黄(Curcuma longa)根茎中提取的一种酚性色素,具有抗菌、抗肿瘤及抗氧化作用。本研究旨在探讨姜黄素对人鼻咽癌CNE-2Z细胞侵袭和转移的影响,并探讨其可能的机制。方法:以10、20、40和80μmol/L姜黄素分别处理CNE-2Z细胞24、48 h后,MTT法检测其对细胞的生长抑制作用。应用Transwell小室进行人工重组基底膜(matrigel)侵袭和运动实验,观察姜黄素对CNE-2Z细胞侵袭和转移的影响。RT-PCR和Western blot分别检测不同浓度姜黄素作用后细胞中表皮生长因子受体(epidermal growth factor receptor,EGFR)表达水平的变化。结果:应用姜黄素处理后,人鼻咽癌CNE-2Z细胞生长受到抑制,且作用呈时效-量效依赖关系;同时细胞侵袭和迁移能力明显降低;EGFR基因和蛋白表达亦明显减弱。结论:姜黄素能够减弱人鼻咽癌CNE-2Z细胞的体外侵袭和转移能力,其机制可能与降低EGFR的表达有关。     

姜黄素诱导低分化鼻咽癌细胞株CNE-2Z的凋亡

背景与目的:鼻咽癌(nasopharyngealcarcinoma,NPC)是我国南方地区的高发肿瘤,发病早期的治疗主要是以放疗为主,而对晚期NPC患者则有必要进行适当的化疗。一些药物可通过诱导肿瘤细胞凋亡来达到治疗肿瘤的目的。本研究拟探讨姜黄素对人鼻咽癌细胞株CNE-2Z体外增殖抑制作用及凋亡的影响。方法:不同浓度姜黄素处理CNE-2Z细胞,用噻唑蓝(MTT)法测定增殖抑制率和IC50;用流式细胞术、Hoechest33258/碘化丙啶(PI)双染、琼脂糖电泳法观察细胞凋亡。结果:姜黄素能抑制CNE-2Z细胞的生长,其效果与姜黄素的浓度和作用时间有关,作用24h、48h、72h的IC50值为(24.05±0.47)、(19.20±0.17)、(7.35±0.50)μmol/L;5、10、20μmol/L姜黄素处理细胞24h后,流式细胞术观察到细胞凋亡率分别为(4.9±3.2)%、(10.7±2.7)%和(14.7±0.5)%;10、20μmol/L姜黄素处理细胞24h后,荧光染色可见细胞缩小,染色质固缩,核染色体碎裂等凋亡形态学改变,琼脂糖凝胶电泳可见DNA梯形条带。结论:姜黄素可诱导CNE-2Z细胞凋亡,姜黄素对CNE-2Z细胞具有增殖抑制作用。     

田基黄对人鼻咽癌细胞株CNE-2细胞生长抑制的体外实验

目的：观察田基黄对人鼻咽癌细胞株CNE-2细胞的生长抑制作用，为其临床应用提供理论依据。方法：将浓度为（0、25、100、400）mg／ml的田基黄与CNE-2细胞作用（24、48、72）h，用SRB法检测田基黄对CNE-2细胞的生长抑制率，流式细胞仪分析细胞周期的变化、检测细胞凋亡率，荧光显微镜观察细胞形态学变化。结果：田基黄对CNE-2细胞的生长抑制率随药物浓度和作用时间的增加而增大。流式细胞仪分析显示田基黄将CNE-2细胞明显阻滞于s期。田基黄浓度为400mg／ml作用48小时，CNE-2细胞的凋亡率为13．12％，荧光显微镜下观察有凋亡细胞存在。结论：田基黄能够抑制人低分化鼻咽癌细胞株CNE-2细胞的增殖，能使CNE-2细胞阻滞于S期。可诱导CNE-2细胞发生-定程度的凋亡。     

田基黄对人鼻咽癌细胞株CNE-2细胞生长抑制的体外实验

目的:观察田基黄对人鼻咽癌细胞株CNE-2细胞的生长抑制作用,为其临床应用提供理论依据.方法:将浓度为(0、25、100、400)mg/ml的田基黄与CNE-2细胞作用(24、48、72)h,用SRB法检测田基黄对CNE-2细胞的生长抑制率,流式细胞仪分析细胞周期的变化、检测细胞凋亡率,荧光显微镜观察细胞形态学变化.结果:田基黄对CNE-2细胞的生长抑制率随药物浓度和作用时间的增加而增大.流式细胞仪分析显示田基黄将CNE-2细胞明显阻滞于S期.田基黄浓度为400mg/ml作用48小时,CNE-2细胞的凋亡率为13.12%,荧光显微镜下观察有凋亡细胞存在.结论:田基黄能够抑制人低分化鼻咽癌细胞株CNE-2细胞的增殖,能使CNE-2细胞阻滞于S期,可诱导CNE-2细胞发生一定程度的凋亡.     

CpG ODN107增强人鼻咽癌CNE-2细胞对β射线照射的敏感性

目的：筛选能提高鼻咽癌CNE-2细胞对β射线放射敏感性的CpGODN序列，观察筛选获得的CpGODN序列放射增敏的作用特点，为寻找新的鼻咽癌放疗增敏剂提供实验依据。方法：应用MTT实验筛选21种CpGODN序列中对人鼻咽癌CNE-2细胞放射增敏作用最强的序列，以MTT实验、集落形成实验、划痕实验和流式细胞术检测该最强作用序列与β射线联合作用对CNE-2细胞的增殖、集落形成、细胞迁移、细胞周期与凋亡的影响。结果：筛选实验显示CpGODN107对CNE-2细胞具有最高的增敏比（1．59&#177;0．06），CpGODN107和β射线联合作用能以剂量依赖方式显著抑制CNE-2细胞的增殖，抑制效应显著高于单纯照射（P〈0．05，P〈0．01）；联合作用显著抑制CNE-2细胞集落形成和迁移能力，显著诱导细胞周期阻滞于G0／G1期，显著增加细胞凋亡（P〈0．01），上述作用效应均明显强于单纯B射线照射（P〈0．05或P〈0．01）。结论：CpGODN107可显著增强人鼻咽癌CNE-2细胞对β射线的照射敏感性，其放射增敏作用可能与诱导细胞周期阻滞和凋亡有关。     

模拟IMRT模式下相对剂量率变化对CNE-2细胞周期及Cyclin D1、Cyclin B1蛋白表达的影响

目的探讨模拟调强放射治疗（intensity-modulated radiation therapy IMRT）对人鼻咽低分化鳞癌细胞株（CNE-2）细胞周期及Cyclin D1、Cyclin B1蛋白水平的影响。方法选取CNE-2为实验对象,分为空白对照组、急速照射组（acute radiation AR）及模拟IMRT（simulational intensity-modulated radiation SIMR）组,后2组分别给予6MV-X线2、4、6、8 Gy 4个剂量点的照射。急速照射组完成时间1-3分钟;模拟IMRT组：各个剂量点分别等分分割5次,每次间隔8-8.5分钟,总时间35分钟照射完成。应用流式细胞术（FCM）分析细胞周期再分布的差异;应用Western blotting检测细胞周期相关因子Cyclin B1、Cyclin D1的蛋白水平。结果在相同剂量点,模拟IMRT组G2期细胞比例低于急速照射组,G2期细胞比例随照射剂量的增加而逐渐增加。急速照射与模拟IMRT组不同剂量点之间以及在相同剂量点比较,Cyclin D1的蛋白表达差异均无统计学意义（P值均〉0.05）;Cyclin B1的蛋白表达差异均有统计学意义（P值均〈0.05）,且随照射剂量的增加而下降,模拟IMRT组Cyclin B1的表达高于急速照射组。结论模拟IMRT照射时间延长,对G2期阻滞作用下降,细胞周期素Cyclin B1的表达相对升高。     

Suppression of BCL-2 synergizes cisplatin sensitivity in nasopharyngeal carcinoma cells

The efficacy of cisplatin for treating nasopharyngeal carcinoma (NPC) is limited by the dose-related toxicities and the development of resistance to cisplatin. Recent studies have shown that B cell lymphoma-2 (BCL-2) is overexpressed and confers chemoresistance in NPC. Thus, targeted therapy against BCL-2 may enhance the antitumour effects of chemotherapy by sensitizing the tumor cells to undergo apoptosis. This study evaluated the combined effects of BCL-2 inhibition and cisplatin in NPC cells. Our results demonstrate that inhibition of BCL-2 by small-hairpin RNA (shRNA) or the BCL-2 inhibitor YC137, synergizes cisplatin sensitivity in NPC cells that overexpress BCL-2. We also show that YC137 enhance cisplatin-induced apoptosis in HK1 and CNE1 cells through suppression of BCL-2 protein expression, induction of mitochondrial depolarization and activation of caspase 9 and caspase 3/7. These findings suggest that the combination of BCL-2 inhibition and cisplatin represents a promising strategy for treating NPC.     

Adenovirus-p53 gene therapy in human nasopharyngeal carcinoma xenografts.

BACKGROUND: One major challenge to human cancer gene therapy, is efficient delivery of the gene-vector complex. METHODS AND RESULTS: Using two distinct human nasopharyngeal carcinoma (NPC) models, we demonstrate that intra-tumoural (IT) administration of adenoviral-mediated wild-type p53 gene therapy (Ad-p53) caused no greater inhibition of tumour growth as compared to ionizing radiation (XRT) alone. Detailed histologic examination of tumour sections demonstrated that <15% of tumour cells were transduced by IT adv-beta-gal. CONCLUSIONS: This report underscores the importance of developing gene transfer vectors, which can provide therapeutic levels of transgene expression efficiently in solid tumours.     

大黄素乙酰化物致鼻咽癌 CNE-1 细胞线粒体自噬活性的研究

目的研究大黄素乙酰化物作用鼻咽癌CNE-1细胞后,其线粒体损伤及其与线粒体自噬的关系。方法以5mg·L^-1及10mg·L^-1大黄素乙酰化物作用鼻咽癌CNE-1细胞,通过透射电镜观察大黄素乙酰化物导致鼻咽癌CNE-1细胞自噬体形成的超微结构;激光共聚焦显微镜测定线粒体膜电位、细胞内钙离子、细胞内活性氧的浓度变化及线粒体自噬的活性。结果与空白对照组比较,两个实验组细胞都出现线粒体损伤以及线粒体自噬体。大黄素乙酰化物作用后的细胞线粒体膜电位降低（P〈0．05）;细胞内游离的钙离子浓度升高（P〈0．05）;细胞内活性氧增加（P〈0．05）,且呈剂量依赖性。经线粒体与溶酶体示踪分析,可见大黄素乙酰化物作用的实验组均出现酸性溶酶体增殖,其中包含线粒体探针标记的线粒体碎片等典型的自噬特征,且增加的趋势呈剂量效应关系。结论大黄素乙酰化物可致鼻咽癌CNE-1细胞线粒体损伤,并与线粒体自噬密切相关。     

塞来昔布对人鼻咽癌CNE-2细胞生长抑制及放射增敏研究

目的 研究塞来昔布对人鼻咽癌细胞系CNE-2细胞生长影响及有无放射增敏作用.方法 (1)细胞生长抑制研究:用MTT法检测细胞生长抑制,流式细胞术检测细胞周期分布及凋亡,透射电子显微镜观察细胞凋亡形态,SP法检测细胞COX-2表达.(2)放射增敏研究:随机设置照射对照、药物对照、单纯照射、药物+照射组,其中成克隆实验单次照射2、4、6、8、10 Gy,细胞周期分布和凋亡实验单次照射6 Gy.结果 塞来昔布显著抑制CNE-2细胞生长并呈浓度和时间依赖性,IC50为80 μmol/L.细胞周期分布显示G0+G1期细胞显著升高(47.03±2.76:56.17±1.95,t=4.68,P=0.010),而S、G2+M期细胞明显下降(33.07±1.86:24.87±1.76,t=5.54,P=0.010;19.30±0.53:17.73±0.83.t=2.75,P=0.050)并呈浓度依赖性.凋亡率显著增高(1.57±0.47:10.47±0.31,t=27.39,P=0.000)并呈浓度依赖性.电镜观察到细胞皱缩、核质浓缩、核碎裂等凋亡形态学改变.SP法检测塞来昔布显著下调CNE-2中COX-2表达[17.48±0.34、12.82±0.51(t=13.20,P=0.00)].塞来昔布的放射增敏比(D0值比为1.74:1.52)为1.15.4个组别细胞周期分布结果 显示单纯照射、药物+照射组的G2+M期细胞明显高于照射对照、药物对照组(68.00±1.65、54.27±5.74、17.60±0.80、14.86±1.23,t=47.70,P=0.000;t=11.63,P=0.000),且单纯照射与药物+照射组间也不同(t=3.99,P=0.020);单纯照射、药物+照射组的细胞凋亡率也明显高于照射对照、药物对照组(4.83±0.97、9.50±1.35、1.33±0.36、2.28±0.42,t=4.67,P=0.01;t=8.81,P=0.000),且单纯照射与药物+照射组也不同(t=4.85,P=0.010).结论 塞来昔布能抑制人鼻咽癌CNE-2细胞生长和诱导凋亡,其机制可能涉及COX-2依赖途径.塞来昔布还能增强CNE-2细胞放射敏感性,可能机制与抑制放射后亚致死损伤修复、直接抑制细胞增殖和增强细胞对放射诱导凋亡率有关.     

塞来昔布对人鼻咽癌CNE-2细胞生长抑制及放射增敏研究

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.     

两种人类鼻咽癌细胞系CNE—1 CNE—2Z体外侵袭过程的定量分析

Computer-image processing system was used in the quantitative study on invasion in organ culture of two human nasopharyngeal carcinoma cell lines of different degree of differentiation, CNE-1 and CNE-2Z. For invasion of tumor cells into precultured heart fragments (PHF), observations were done on days 1,3,5 and 7 after organ culture. The results showed that the invasion of both CNE-1 and CNE-2Z cells was progressive. The invasion of CNE-1 tumor cells covered 82.22% of PHF on day 7, but on days 1 to 3 the invasion of CNE-2Z tumor cells covered 80-92% of PHF. This indicates that CNE-2Z cells are more invasive then CNE-1 cells.     

塞来昔布对人鼻咽癌CNE-2细胞生长抑制及放射增敏研究

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.     

塞来昔布对人鼻咽癌CNE-2细胞生长抑制及放射增敏研究

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.     

塞来昔布对人鼻咽癌CNE-2细胞生长抑制及放射增敏研究

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.     

塞来昔布对人鼻咽癌CNE-2细胞生长抑制及放射增敏研究

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.