人永生化食管上皮细胞恶性转化的验证

Malignant transformation of the immortalized esophageal epithelial cells

目的 检查永生化人胚食管上皮细胞系 (SHEE)的恶性表型、成瘤性和侵袭力 ,证实此细胞系传至 85代 (SHEE85 )已完全恶性转化。方法 培养的SHEE85细胞用光镜和电镜观察细胞形态 ;以流式细胞仪检查细胞周期 ;行细胞软琼脂培养 ;以裸小鼠和SCID小鼠接种检测其成瘤性 ;以细胞培养于羊膜的体外方法和移植至SCID小鼠腹腔的体内方法测定其侵袭力。结果 SHEE85在光镜下细胞生长拥挤 ,大小不一 ;电镜下见细胞呈增殖状态。DNA组方图统计细胞增殖指数为 4 7% ,高倍体细胞 12 %。软琼脂培养多细胞大集落形成率 4 %。接种的 4只裸小鼠和 4只SCID小鼠皆成瘤。结论 SHEE85已完全恶性转化 ,并且有较强侵袭力。

Objective Immortal cell line of human embryonic esophageal epithelium (SHEE) was induced by E6E7 genes of human papillomavirus (HPV) type 18 in our laboratory. To identify the fully malignant transformation at its 85th passage (SHEE85), the malignant phenotype, tumorigenesis and invasive potency were studied. Method The cultured SHEE85 cells were observed under the light and the electron microscope (EM) for cell morphology, analyzed by flow cytometry for cell cycle. The tumorigenesis was assayed by plating cells in soft agar and transplanting cells into the nude mice and SCID mice. To detect invasive potency, cells were cultured on amniotic membrane in vitro and transplanted into peritoneal cavity of mice in vivo .Results SHEE85 cells were crowded in cultivation with different sizes and shapes under light microscope, and displayed proliferative morphology under EM. Proliferative index was 47% with 12% hyperploidy cells in determination of DNA histogram. Many large colonies grew in soft agar (4%) and the transplanted tumors were found in all 4 nude and 4 SCID mice, with strong invasive potency demonstrated in vitro and in vivo .Conclusion The immortal esophageal epithelial cell line induced by HPV18 E6 E7 is derived from a fully malignant transformation with a strong invasive potency at the 85th passage. It is also a reliable model for studying the cellular and molecular mechanisms of carcinogenesis of the esophageal carcinoma. [