BCSC-1基因异位表达抑制鼻咽癌细胞的恶性生物学行为

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。

Inhibition of malignant activities of nasopharyngeal carcinoma cell by ectopic expression of BCSC-1 gene

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.