BCSC-1基因异位表达抑制鼻咽癌细胞的恶性生物学行为  

Inhibition of Malignant Activities of Nasopharyngeal Carcinoma Cell by Ectopic Expression of BCSC-1 Gene

作　　者：陈双玲[1] 周异群[1] 田云[1] 鞠吉雨[1] 刘音[1] 朱立平[1] 

机构地区：[1]中国医学科学院北京协和医学院基础医学研究所免疫学系,北京100005

出　　处：《中国医学科学院学报》2007年第5期612-617,共6页Acta Academiae Medicinae Sinicae

基　　金：国家重点基础研究发展计划项目(973计划)(2001CB510004);家高技术研究发展计划项目(863项目)(2002BA711A03)~~

摘　　要：目的研究BCSC-1基因异位表达对人鼻咽癌细胞CNE-2L2恶性生物学行为的影响。方法RT-PCR扩增BCSC-1cDNA,分别构建原核重组表达载体pMAL-c2X-BCSC-1和真核重组表达载体pcDNA4/myc-HisA-BCSC-1。在原核细胞表达BCSC-1蛋白。用BCSC-1蛋白免疫新西兰白兔,制备BCSC-1抗血清。采用实时定量RT-PCR与用BCSC-1抗血清免疫荧光染色确认野生型CNE-2L2细胞的BCSC-1基因低表达。藉脂质体把pcDNA4/myc-HisA-BCSC-1转染入野生型CNE-2L2细胞。Westernblot检测BCSC-1基因在野生型CNE-2L2细胞异位表达。用体外培养中的生长曲线和集落生成检测细胞生长。接种细胞于裸鼠,定时测量肿瘤大小。小鼠肺做连续组织切片,观察肿瘤的肺转移。结果制备了BCSC-1抗血清。明确BCSC-1基因在野生型CNE-2L2细胞的表达极低。制备了BCSC-1基因异位表达的CNE-2L2细胞。与对照细胞(野生型CNE-2L2细胞和转染pcDNA4/myc-HisA的CNE-2L2细胞)相比,BCSC-1基因异位表达的CNE-2L2细胞在培养中的生长、集落生成、细胞的成瘤性生长和所长肿瘤的肺转移均明显抑制。结论BCSC-1基因异位表达对CNE-2L2细胞的恶性生物学行为有明显抑制作用。Objective To study effects of ectopic expression of BCSC-1 gene on the malignant activities of human nasopharyngeal carcinoma cell CNE-2L2. Methods BCSC-1 cDNA was isolated by RT-PCR and inserted into pMAL-c2X and pcDNA4/myc-His A. BCSC-1 protein was expressed in prokaryocytes. Rabbit antiserum to BCSC-1 was developed by means of immunization of rabbit with the BCSC-1 protein. Expression of BCSC-1 gene in wild type CNE-2L2 cell （ W cell） was examined by real-time RT-PCR and immunofluorescence staining with the antiserum as a probe, pcDNA4/myc-His A-BCSC-1 was transfected into W cell at the presence of LipofectAmine. The cells were selected by G418 and cloned. Ectopic expression of BCSC-1 gene in W cell was examined by Western blot. Cell growth was detected by drawing of growth curves and colony formation tests. Cells were inoculated into nude mice. Size of tumors was assayed once a week. Lungs of the mice were sectioned continuously and metastatic loci in lungs were examined upon a microscope. Results Rabbit BCSC-1 antiserum was prepared. Expression of BCSC-1 gene in W cell was found to be very low. CNE-2L2 cell with ectopic expression of BCSC-1 gene was developed. Growth in vitro, colony formation, tumorigenesis in nude mice, and lung metastasis of the tumor were profoundly inhibited of the cell with ectopic expression of BCSC- 1 gene in comparison with controls, wild type cell and the cell transfected with mock. Conclusion Ectopic expression of BCSC-1 gene exerts profound inhibitive effect on the malignant activities of CNE-2L2 cell.

关 键 词：BCSC-1基因 鼻咽癌 生长曲线 集落生成 成瘤性生长 转移 

分 类 号：R739.63[医药卫生—肿瘤]