人乳头状瘤病毒诱导人胚食管上皮永生化细胞恶性转化

为了证实ＨＰＶ１８Ｅ６Ｅ７基因诱导的人胚食管上皮永生化细胞（ＳＨＥＥ）６１代（ＳＨＥＥ６１）部份细胞（ＳＨＥＥ６１Ａ）已经恶性转化，以寻找监控细胞早期恶性转化的方法。对培养的ＳＨＥＥ第１０代（ＳＨＥＥ１０）和６１代（ＳＨＥＥ６１）细胞，用光学是为微镜和电子显微镜观察细胞形态及生长形式；用流工细胞仪分析细胞周期；用染色体Ｇ带核型分析和间期核染色体１、７、８号着丝粒探针荧光原位杂交９ＦＩＳＨ）检测细胞

Malignant Transformation of Immortalized Human Embryonic Esophageal Epithelial Cells lnduced by Human Papillomavirus

This paper intented to identify the inducement of malignant transformation in parts of immortalized human embryonic esophageal epithelial cells (SHEE) by HPV18 E 6E 7 at the 61th passage and to search a method to monitor the early malignant transformation of the cells. The cultured SHEE cells at its 10th(SHEE10) and 61st passages (SHEE 61) were observed under light and electron microscope for cell morphology and cell growth pattern, analyzed by flow cytometry for cell cycle, tested by chromosome G-band idiogram and assayed for centromeres of the interphase nucleus with the probes of chromosomes 1,7,8, by FISH. The rapidly growing cell clone (SHEE61A) was selected with optimum seeking method from SHEE61 cell line and it was used to inoculate into nude mice and SCID mice. The results showed that SHEE61 cells were different in size and shape and grew overlapped under light microscope. They showed a proliferative phenotype with polymorphic nuclei and enlarged nucleoli under electron microscope. SHEE10 and SHEE61 cells when tested by flow cytometry, both had the similar proliferative cycle as indicated by DNA histograms, but the proliferative index and the cells with DNA>4n in SHEE61 cells were more than that in SHEE10 cells. Chromosome modal number of SHEE61 cells revealed two subpopulation, 57-60 and 63-65, and in chromosomes 1,7,9,13,17, the trisomy, tetrasomy and pentasomy were frequently seen. FISH revealed the centromere increment in interphase nuclei of chromosomes 1,7. When both SHEE 10 and SHEE61A cells were separately inoculated into SCID mice, the latter developed tumors and infiltrated into the muscular layer. So, it can be judged that SHEE61A cells has been malignantly transformed. The experimental data showed that the early malignantly transformed cells may be monitored by examining cell genetic characteristics and the clone optimum seeking method.