高剂量X射线照射对人鼻咽癌CNE-1细胞生物学特性影响的研究  

Effect of high-dose X-ray exposure on the biological characteristics of human nasopharyngeal carcinoma cells line CNE-1

作　　者：宋学薇[1] 皇婷[1] 阙开乾[1] 王若雨[1] 王秀丽[2] 

机构地区：[1]大连大学附属中山医院肿瘤科,辽宁大连116001 [2]中国科学院大连化学物理研究所,辽宁大连116023

出　　处：《中华肿瘤防治杂志》2012年第10期721-725,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30970869)

摘　　要：目的:分别以照射前的人鼻咽鳞癌CNE-1细胞和高剂量X射线(50Gy)照射后的CNE-1细胞为研究对象,探讨高剂量X射线照射后的CNE-1细胞的某些生物学特性的变化。方法:采用细胞毒实验(SRB法)检测两组细胞在化疗药物作用下的IC50值,RT-PCR测两组细胞MDR-1基因,共聚焦显微镜测P-gp蛋白,对比两组细胞的耐药性;MTT法测定两组细胞生长曲线,共聚焦显微镜检测Ki-67蛋白,比较两组细胞的生长增殖能力;采用细胞黏附实验比较两组细胞的黏附性,RT-PCR检测两组细胞PECAM-1基因的表达,探讨PECAM-1和细胞黏附的相关性。结果:照射后CNE-1细胞在多柔比星作用下的IC50值是照射前细胞的1.8倍(P=0.004),在紫杉醇作用下的IC50值是照射前细胞的5.3倍(P=0.003),两组细胞MDR-1基因半定量值分别为0.17±0.01和0.34±0.04(P=0.002),照射后细胞P-gp蛋白高表达于照射前细胞;在对数生长期时,照射后细胞生长快于照射前细胞,且其Ki-67蛋白表达高于照射前细胞;照射后细胞在各时间点的黏附率都明显高于照射前细胞,两组数值比较差异有统计学意义,P<0.05;照射前后两组细胞PECAM-1基因的半定量值分别为0.40±0.02和0.51±0.01,P=0.002。结论:高剂量X射线照射后的CNE-1细胞产生耐药性;生长增殖能力及黏附特性高于照射前细胞,PECAM-1基因也高表达,Ki-67及PECAM-1均可能参与耐药。OBJECTIVE：To explore the effect of high-dose X-ray exposure on the growth and phenotype of the human nasopharyngeal carcinoma cell line CNE-1. METHODS： SRB assay was used to measure IC50 values of the anti-cancer drugs （Adriamycin and Taxol） in the X-ray treated CNE 1 cells. Their growth was assayed by MTT assay. Non-treated CNE1 cells served as control. To clarify the phenotype change of the CNE-1 cell before and after X-ray treatment,ex pression of P-glycoprotein （P-gp） and Ki-67 was evaluated through immunofluorescence staining. Confocal microscopy was used to capture the images. Multidrug resistance gene （MDR-1） and platelet endothelial cell adhesion molecule （PE- CAM-1） gene expression was detected by RT-PCR. RESULTS： The IC50 values of ADM and Taxol in the X-ray treated CNE1 cells was 1.8 times and 5.3 times higher respectively than those of the non-treated cells （control）, P= 0. 004, P= 0. 003. The growth rate of the treated cells was improved obviously as evidenced by the growth curve. Moreover, expression of MDR 1 gene and PECAM-1 gene in the treated cells increased significantly when compared with the control. The gene semi-quantitative assessment of MDR-1 was 0.17±0.01 and 0.34±0.04（P=0. 002） ,while that of PECAM-1 was 0.40±0.02 and 0. 51±0. 01（P=0. 002）. The adhesion capability of the CNE1 cells after X-ray treatment was also enhanced significantly （P〈 0. 05）. CONCLUSION： High-dose X-ray exposure can lead to some alteration of biological characteristics of CNE-1 cells,including enhanced cell proliferation,improved drug resistance and cell adhesion.

关 键 词：鼻咽肿瘤 耐药性 黏附 KI-67 PECAM-1 

分 类 号：R739.63[医药卫生—肿瘤]