定量研究EB病毒潜在膜蛋白对人鼻咽癌细胞系CNE1的作用

MORPHOLOGICAL QUANTITATIVE STUDY ON HUMAN NASOPHARYNGEAL CARCINOMA CELL LINE CNE1 TRANFECTED WITH EB VIRUS LMP GENE

目的研究ＥＢ病毒ＬＭＰ对人高分化鼻咽癌细胞株ＣＮＥ１细胞形态及ＤＮＡ含量的影响。方法采用电穿孔基因转染技术，将重组ＥＢＶ┐ＬＭＰ表达质粒转染鼻咽癌细胞株ＣＮＥ１细胞。以载体质粒转染及ＣＮＥ１细胞为对照，用免疫荧光和图像分析方法，观察细胞ＬＭＰ表达、细胞形态及ＤＮＡ含量的变化。结果ＬＭＰ表达质粒转染后细胞浆及细胞膜ＬＭＰ强阳性表达。形态定量及ＤＮＡ含量测定表明，ＬＭＰ表达后，ＣＮＥ１细胞明显变小（Ｐ＜０．０１），核浆比显著增大（Ｐ＜０．０１），核浆的变化主要由胞浆面积的增减所致，而与核面积的改变则无明显正比关系；ＤＮＡ含量明显增高（Ｐ＜０．０１／０．０５），且倍体分布更分散而右移。结论ＬＭＰ对ＣＮＥ１细胞生长有明显的促进作用，可致细胞恶性程度增高且分化受抑，有向低分化鳞癌发展倾向。

Objective:Effects on human nasopharyngeal carcinoma (NPC) cell line CNE1 transfected by EBV LMP gene were studied.Methods:Electroporation was used to transfect LMP gene and the vector into CNE1 cells. Immunofluorescence technique (IFT) and image analysis technique were used. Results::LMP gene was expressed stably on NPC CNE1 cells after transfection.The LMP expressive CNE1 cell showed that cytoplasmic area reduced significantly ( P <0 01) with a higher nuclear/cytoplasm ratio( P <0 01),DNA content increased sufficiently ( P <0 01/0.05),chromosomal ploidy presented a wider distribution and shifted to right.The nuclear/cytoplasm ratio change chiefly caused by the reduction of cytoplasm area,not by the nuclear area.Conclusion:The EBV LMP can promote the growth of NPC CNE1 cells,resulted in higher malignancy and poorer differentiation.The CNE1 cells tends to change to low differentiated squamous cell carczinoma.These results indicated that LMP may have an important role on the progression of NPC cells.