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为了研究病毒和促癌物在食管癌形成中的作用，用带有人乳头状瘤病毒１８型Ｅ６Ｅ７片段的载体腺病毒（简称ＨＰＶ１８Ｅ６Ｅ７ＡＡＶ）感染人胚食管上皮细胞，然后加ＴＰＡ协同作用，观察细胞转化。将人胚食管切碎，与ＨＰＶ１８Ｅ６Ｅ７ＡＡＶ同孵育２小时，在加有１０％小牛血清的１９９培养液培养和传代，形成永生化细胞株，即人胚食管上皮细胞汕头株（ＳＨＥＥ）。实验分两组：一组ＳＨＥＥ细胞在传代至第５和１３代时，两次在培养基中加入ＴＰＡ（１２Ｏｔｅｔｒａｄｅｃａｎｏｙｌｐｈｏｒｂｏｌ１３ａｃｅｔａｔｅ）５ｎｇ／ｍｌ，每次诱导２周，所获得的细胞株称为人胚食管上皮癌细胞汕头株１号（ＳＨＥＥＣ１）；另一组ＳＨＥＥ细胞培养条件相同，未加ＴＰＡ，为对照组。细胞转化的形态表型由光学显微镜、电子显微镜和荧光显微镜检查；ＤＮＡ含量和细胞周期用流式细胞仪检测；用３５ｍｍ软琼脂培养皿接种１０３细胞（第２０代），每组５碟，计算集落形成率；裸鼠皮下接种１０６细胞检测致瘤性；用荧光原位杂交（ＦＩＳＨ）和ＰＣＲ检测ＨＰＶ１８Ｅ６Ｅ７。结果表明：细胞ＤＮＡ合成和增殖指数（ＰＩｘ），ＳＨＥＥＣ１组（４５％）高于ＳＨＥＥ组（３４％）；高倍体细胞数，ＳＨＥ?
To investigate the role of viruses and carcinogens in esophageal carcinogenesis, human embryonic esophageal epithelial cells were infected with a vector adenovirus carrying human papillomavirus type 18 E6E7 (abbreviated HPV18E6E7AAV) and then synergized with TPA to observe cell transformation . The human embryo esophagus was shredded, incubated with HPV18E6E7AAV for 2 hours, and cultured and passaged in 199 medium supplemented with 10% fetal bovine serum to form an immortalized human cell line SHEE. The experiment was divided into two groups: a group of SHEE cells in passage to the 5th and 13th generation, twice in the medium by adding TPA (12 O tratecanoylphorbol 13 acetate) 5ng / ml, each induction for 2 weeks, The obtained cell line was named human embryonic epithelial carcinoma cell line Shantou No.1 (SHEEC1). The other group of SHEE cells were cultured in the same condition without TPA as the control group. Morphology of cell transformation was examined by light microscopy, electron microscopy and fluorescence microscopy; DNA content and cell cycle were measured by flow cytometry; 103 cells (passage 20) were seeded in 35 mm soft agar plates with 5 dishes in each group and calculated The rate of colony formation was established. The nude mice were subcutaneously inoculated with 106 cells for tumorigenicity detection. HPV18E6E7 was detected by fluorescence in situ hybridization (FISH) and PCR. The results showed that the DNA synthesis and proliferation index (PIx) of SHEEC1 group (45%) was higher than that of SHEE group (34%);