医学检验信息网 检验医学 2007-2-26 16:16:50

【摘要】 目的 SHEE细胞系是经人乳头状瘤病毒(HPV)18型E6E7基因诱导的永生化上皮细胞株，已传代超过50代。研究胎儿食管上皮永生化的细胞株SHEE生物学特性，包括增殖，分化和调亡。方法 细胞于199培养基培养，用光镜、电镜和荧光显微镜研究其生长率、形态和染色体分析；用流式细胞仪研究其细胞增殖动力学；用免疫组织化学方法研究Ki67和角蛋白和用末端转移酶标记(TUNEL)凋亡细胞。结果 细胞培养呈单层，锚定依赖和接触抑制生长。细胞生长曲线，增殖期3～8天，平顶期9～10天，衰亡期10天以后。增殖指数(PIx)34.0%，分裂指数(MI)2.47%(1.20%～4.80%)，凋亡指数(AI)1.30%～6.90%，核染色体众数46条为主(44～54条)和细胞周期DNA呈2倍体核型分布。电镜可见胞浆有张力原纤维，角蛋白免疫组织化学阳性证实鳞状上皮有分化特征。细胞凋亡可发生在增殖期，特别在衰亡期增多。结论 从生物学行为看来，SHEE细胞株接近于其来源细胞，胎儿食管上皮基底层，保存其增殖能力和分化的潜力。细胞死亡(包括细胞凋亡)是细胞群体扩增调节重要的因素，应作为研究生物学特性内容之一。 Biological characteristics of human fetal esophageal epithelial cell line immortalized by the E6 and E7 gene of HPV type 18

SHEN Zhongying, SHEN Jian, ZENG Yi, et al.

(Medical College of Shantou University Shantou 515031_

【Abstract】 Objective Biological features including proliferation, differentiation and cell death of SHEE cell line, an immortalized epithelium of the fetal esophageal epithelium induced by HPV 18 E6E7 AAV, were studied.Methods SHEE cell line being cultured for more than 50 passages, were cultured in 199 growth medium and were examined by light, electron and fluorescence microscopy for growth rate, morphological features and chromosome analysis, by flow cytometry for cell proliferative dynamics, by immunohistochemistry for Ki67 and cytokeratin, and by terminal DNA transferase label (TUNEL) for apoptosis.Results At the 20th passage, the SHEE cell remained monolayer, anchorage-dependent and attachment-inhibited growth. The growth curve showed proliferative phase 3～8th days, top-plate phase 9～10th days and attenuative phase after 10th days. Proliferative index(PIx)34.0%, mitotic index (MI)2.74%(1.20%～4.80%), apoptotic index (AI)1.30%～6.90%, chromosome analysis mainly 46 (44～54/nucleus) and DNA distribution in diploidy were calculated and described. The tonofilament expression in cell cytoplasm by electron-microscopy and positive reaction of cytokeratin by immunochemistry showed differentiative character of squamous epithelium. The cell apoptosis occured in the proliferative phase and especially increased in attenuative phase.Conclusion Of biological behaviors, the SHEE cells are close to the basal cells of their original fetal esophageal mucosa keeping proliferative and differentiative potency. This study suggests that the cell death (including cell apoptosis) may be an important factor in studying of cell growth regulation and it may be an research area for cellular biological behaviors. 【Key words】 Fetal esophageal epithelium Immortalization Biological behaviors HPV 18 E6E7 AAV 细胞在体外培养研究其生物学特征，主要是观察其生命行为，即增殖、分化、死亡(包括凋亡)，寻找其特点和生长规律。细胞离体进行培养，虽然其原本细胞遗传学特点仍然保留，但因外界环境改变，失去体内多种细胞和神经体液的影响，细胞的各种特性的表型有所改变。经多次传代细胞基因组不稳定，产生表型差异^［1］，致使体内细胞和体外培养细胞生物特性的差异。本研究用人乳头状瘤病毒(HPV)18型E6E7AAV感染，胎儿食管上皮诱导永生化，建立细胞系SHEE^［2］，已传代超过50代。取冻存20代SHEE细胞，观察其增殖，分化和死亡等指标，研究SHEE细胞系的生物学特性。 1 材料和方法

责任编辑: 参与评论