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目的： 探讨上调ｐ１６ 基因的表达对鼻咽癌细胞恶性表型逆转的作用， 为ｐ１６ 基因在鼻咽癌发病过程中具有一定的作用提供直接的证据。方法： 本文采用电穿孔的方法向ｐ１６ 基因表达下调的鼻咽癌细胞系ＨＮＥ１ 中导入全长野生型的ｐ１６ ｃＤＮＡ，通过Ｇ４１８ 筛选获得可以稳定传代的转染细胞系。对其中４ 个转染细胞系以ＰＣＲ 和Ｎｏｒｔｈｅｒｎ 杂交鉴定转染细胞系中外源ｐ１６ ｃＤＮＡ 整合以及ｐ１６ 基因表达的状况。并借助比较细胞倍增时间，流式细胞计数和裸鼠接种的方法对转染细胞的恶性表型进行了检测。结果：在所鉴定的４ 个转染细胞系中有一个转染细胞系（ ＃ ４－ ２） 不仅整合了外源ｐ１６ ｃＤＮＡ，且ｐ１６ 基因的表达明显增高。与未转染的鼻咽癌细胞系ＨＮＥ１ 及未整合外源ｐ１６ ｃＤＮＡ 的转染细胞系相比，＃ ４ － ２ 细胞的倍增时间延长，停滞于Ｇ１ 期的细胞数明显增多，接种裸鼠后肿瘤形成的潜伏期延长。结论：以上实验结果证实ｐ１６ 基因在鼻咽癌的发生过程中确实具有一定的作用，它的表达上调可以促进鼻咽癌细胞恶性表型的逆转。
Objective: To investigate the effect of upregulation of p16 gene expression on the reversal of malignant phenotype in nasopharyngeal carcinoma cells and provide direct evidence for the role of p16 gene in the pathogenesis of nasopharyngeal carcinoma. Methods: The full-length wild-type p16 cDNA was introduced into the nasopharyngeal carcinoma cell line HNE1 with down-regulated p16 gene expression by electroporation. Transfected cell lines that could be passaged stably were obtained by G418 selection. The status of exogenous p16 cDNA integration and p16 gene expression in transfected cell lines was identified by PCR and Northern hybridization on 4 transfected cell lines. The malignant phenotypes of the transfected cells were detected by comparing the cell doubling time, flow cytometry and nude mice inoculation. RESULTS: One of the four transfected cell lines identified (#4-2) not only integrated exogenous p16 cDNA, but also significantly increased the expression of the p16 gene. Compared with untransfected nasopharyngeal carcinoma cell line HNE1 and transfected cell line without integrated exogenous p16 cDNA, the doubling time of #4-2 cells was prolonged, and the number of cells arrested at G1 phase was significantly increased, after inoculation of nude mice. The incubation period for tumor formation is prolonged. Conclusion: The above experimental results confirm that p16 gene has a certain role in the development of nasopharyngeal carcinoma, and its up-regulation can promote the reversal of the malignant phenotype of nasopharyngeal carcinoma.