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目的:研究EBVLMP对人高分化鼻咽癌细胞株CNE1体内外增殖能力的影响。方法:以人高分化鼻咽癌细胞株(CNE1)为对象,采用电穿孔基因转染技术,将重组EBVLMP表达质粒转染CNE1细胞。以载体质粒转染及CNE1细胞为对照,用细胞体外增殖实验、细胞软琼脂克隆形成率测定、流式细胞术、PCNA检测和裸鼠成瘤实验,观察细胞增殖能力的变化。结果:结晶紫染色法检测体外细胞增殖,结果表明,A比值实验组高于空白组及阴性对照组(P<001);实验组细胞软琼脂克隆形成率显著高于空白组及阴性对照组(P<001);FCM法测定细胞周期,实验组S期细胞比空白组及阴性对照组显著增高,S期细胞达349%;PCNA免疫组化染色,实验组细胞PCNA阳性率明显高于空白组及阴性对照组(P<001)。CNE1及转染细胞系裸小鼠移植结果表明,LMP表达细胞系移植瘤潜伏期、体内倍增时间明显缩短(P<001),成瘤率、瘤重显著增高(P<001/005)。结论:EBVLMP对CNE1细胞体内外增殖能力有明显的促进作用,提示LMP在NPC细胞演进过程中可能起重要作用,为进一步深入研究提供了依据。
Objective: To investigate the effect of EBVLMP on the proliferation of human highly differentiated nasopharyngeal carcinoma cell line CNE1 in vitro and in vivo. Methods: The human EBVLMP gene was transfected into CNE1 cells using human epidermal growth factor (CNE1) cell line by electroporation. The transfected cells were transfected with CNE1 cells and transfected with CNE1 cells. The cell proliferative ability was observed by cell proliferation assay, soft agar colony formation assay, flow cytometry, PCNA assay and tumorigenesis in nude mice. Results: Crystal violet staining was used to detect cell proliferation in vitro. The results showed that the A ratio in experimental group was higher than that in blank group and negative control group (P <001). The soft agar colony formation rate in experimental group was significantly higher than that in blank group and negative control group P <001). The cell cycle was determined by FCM. The number of S phase cells in experimental group was significantly higher than that in blank group and negative control group, reaching 349% in S phase. The positive rate of PCNA in experimental group was significantly higher than that in blank group And negative control group (P <001). The results of nude mice transplanted with CNE1 and transfected cell lines showed that the LMP-expressing cell line transplanted tumor had a shorter incubation period and shorter doubling time (P <001). The tumorigenicity and tumor weight were significantly increased (P <001/005). CONCLUSION: EBVLMP can significantly promote the proliferation of CNE1 cells in vitro and in vivo, suggesting that LMP may play an important role in the progression of NPC cells and provide a basis for further study.