摘要
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 explore the role of the p16 gene up regulation on the malignant phenotype reversion of nasopharyngeal carcinoma (NPC) cell,and to provide the direct evidence that the p16 gene play a role in nasopharyngeal carcinogenesis. Methods:Full length wild type p16 cDNA was introduced into NPC cell line HNE1 with p16 gene down regulation by electroporation.Stable G418 resistant clones were isolated.Then PCR was used to detect the integration of the exogenous p16 cDNA and Northern blotting was used to detect the expression of the p16 gene in 4/7 clones.Finally the growth suppression of the positive clone (#4 2) was detected by doubling time,cytometry and xenografts. Results:In 4 clones identified, only one transfectant #4 2 showed integration of exogenous p16 cDNA and up regulation of p16 mRNA expression compared to NPC cell line HNE1 and the transfectant clones without integration of exogenous p16 cDNA, transfectant #4 2 showed:a). a longer doubling time; b). an accumulation of cells in G1 phase and c). longer latent period of tumor formation in nude mice. Conclusions:These data show that up regulation of p16 expression can inhibit the growth and tumor formation of NPC cell. The p16 gene is really a functional tumor suppressor gene in nasopharyngeal carcinogenesis and takes part in promoting the malignant phenotype reversion of NPC cells.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
1999年第5期499-503,共5页
Chinese Journal of Cancer
基金
国家杰出青年科学家基金!( 批准号:39525022)
中华医学基金!(CMB)( 批准号:90 - 529) 资助
关键词
P16基因
癌细胞
表达上调
表型
鼻咽肿瘤
p16 gene
NPC cell line
Up regulation
Malignant phenotype reversion
