摘要
目的:建立辐射诱导的人鼻咽鳞癌CNE1/R细胞系,检测缺氧诱导因子-1α(HIF-1α)和多药耐药基因(MDR1)及P-糖蛋白(P-gp)的表达。方法:采用直线加速器对体外培养进入指数增长期的CNE1细胞进行外照射,输出剂量率2Gy/min,2Gy/次,隔日1次,3次/周,累积总剂量50Gy,取照射完成后培养21d的细胞进行检测;以未照射CNE1细胞作为对照。利用RT-PCR检测照射前后HIF-1α及MDR1mRNA的表达;蛋白质印迹法检测照射前后细胞HIF-1α及P-gp蛋白的表达。结果:RT-PCR法检测结果显示,未照射组与照射组鼻咽鳞癌细胞系CNE1中HIF-1αmRNA的半定量值分别为0.23±0.02和0.37±0.04(t=5.42,P=0.01),MDR1mRNA的半定量值分别为0.17±0.01和0.54±0.04(t=15.54,P=0.00),照射组较未照射组明显增高,P<0.05;蛋白印迹法检测结果显示,未照射组与照射组鼻咽鳞癌细胞系CNE1中HIF-1α蛋白的半定量值分别为0.05±0.01和0.08±0.01(t=3.67,P=0.02),P-gp蛋白分别为0.01±0.01和0.04±0.01(t=3.67,P=0.02),照射组较未照射组明显增高,P<0.05。结论:照射组人鼻咽鳞癌CNE1细胞系中HIF-1α和MDR1基因/P-gp蛋白表达均增高,提示核转录因子HIF-1α可能促进细胞内MDR1/P-gp的表达。
OBJECTIVE： To establish human nasopharyngeal CNE1 cell line after high-dose X-rays irradiation（CNE1/R）,and examine the expression of HIF-1α and MDR1.METHODS： The CNE1 cell line were processed with accelerator linear and supplied to X-rays irradiation for 2 Gy/min,every other day for three times in each week,the total dose was 50 Gy.The 21st cultured CNE1/R cell line and CNE1 parent cell line were analyzed.The expression of HIF-1α and MDR1 gene were examined by reverse transcriptional polymerase chain reaction（RT-PCR）;The proteins of HIF-1α and P-gp were detected by Western blotting.RESULTS： Before or after irradiation,the semiquantitative values of HIF-1α genes were 0.23±0.02 and 0.37±0.04（t=5.42,P=0.01）,respectively.MDR1 gene were 0.17±0.01 and 0.54±0.04（t=15.54,P=0.00）,and the semiquantitative values showed a significance difference（P〈0.05）;The semiquantitative values of HIF-1α protein were 0.05±0.01 and 0.08±0.01（t=3.67,P=0.02）,respectively.P-gp protein were 0.01±0.01 and 0.04±0.01（t=3.67,P=0.02）,and the semiquantitative values showed a significance difference（P〈0.05）.CONCLUSION： In CNE1/R cell line,the expression of HIF-1αand MDR1/P-gp are both increased,cuing that the nuclear factor HIF-1α may promote the expression of MDR1/P-gp.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2012年第3期197-200,共4页
Chinese Journal of Cancer Prevention and Treatment
关键词
鼻咽肿瘤
细胞系
辐射
缺氧诱导因子-1Α
多药耐药基因
nasopharyngeal neoplasms
cell line
radiation
hypoxia-inducible factor-1α
multidrug resistance gene
