摘要
目的 观察苦参碱作用于HepG2 细胞后 ,光镜和电镜下细胞形态结构改变以及AFP、PCNA和分裂指数的变化。方法 倒置显微镜下观察活细胞形态 ;光镜下观察Wright和苏丹Ⅲ染色的细胞形态和分裂指数 ;扫描和透射电镜观察细胞表面和内部超微结构改变 ;免疫组化和图像分析 ,检测AFP和PCNA。结果 未用药细胞具有肿瘤细胞典型的形态特征 ,细胞的分裂指数为 5 3 .8‰ ,胞浆内未见透明颗粒和空泡 ,苏丹Ⅲ染色为阴性。免疫组化染色AFP和PCNA为阳性。苦参碱处理细胞后 ,细胞数量减少 ,散在生长 ,形态呈多形性 ,细胞浆内充满圆形、透明的颗粒。Wright染色 ,细胞浆呈空泡状 ,细胞核变小 ,核浆比减小 ,核仁变小、数目减少 ,细胞的分裂指数降低。苏丹Ⅲ染色 ,胞浆内有阳性的颗粒。电镜下 ,细胞表面微绒毛变短、变粗 ,甚至消失。核内异染色质增多 ,核仁变小、甚至消失。细胞浆内滑面内质网扩张 ,并有脂肪空泡和自噬小体形成。胞浆局部还出现了较多的板层状排列的粗面内质网。细胞核膜和胞膜均完整。AFP和PCNA量 ,随用药浓度和作用时间的增加而降低。结论 苦参碱对HepG2 细胞的生长增殖有抑制作用 ,使HepG2 细胞的某些恶性增殖表型减弱或消失 ,反应了该细胞的恶性程度和侵袭力均减弱或消失。
Objective To study the effects of matrine on the cell morphology, cell division index and the contents of alpha fetoprotein (AFP) and proliferating cell nuclear antigen (PCNA) of human hepatocellular carcinoma cell HepG 2 in vitro . Methods: The morphology of the living cells and the cells after Wright or Sudan staining was observed with an inverted and an light microscope respectively. The surface and ultrastructures of the cells were studied with scanning and transmission electron microscope. Index of cell division was obtained with light microscope. The cellular content of AFP and PCNA were measured with immunohistochemical method and image analysis. Results HepG 2 cell without the treatment of matrine showed typical characteristics of cancer cells with an index of cell division of 53.8‰. No transparent granules or vacuoles were found in the cytoplasm. Sudan staining was negative and immunohistochemistry for AFP and PCNA was positive. HepG 2 cells treated with matrine decreased in number and growed in a scattered pattern. They were pleomorphic and their cytoplasm was filled with round and transparent granules. Wright staining revealed vacuoles existed in cytoplasm, the nucleoli were smaller in size, decreased in number and even disappeared and the index of cell division was decreased. Sudan staining showed positive particles in the cytoplasm. It was found with electron microscopy that the microvilli on cell surface became shorter and thicker and even disappeared, heterochromatin was increased in the nuclei, smooth endoplasmic reticulum was expanded and fat vacuoles and autophageosomes were formed, many rough endoplasmic reticula in lamellar arrangement were found in the cytoplasm and both nuclear and cell membranes were intact. The content of AFP and PCNA was independent on the dosage and the duration of matrine administration. Conclusion Matrine can inhibit the growth and proliferation of the HepG 2 cells in vitro and weaken, even eliminate its malignant phenotype. This finding suggests that the malignant degree decreases with the administration of matrine.Consequently, matrine may be a promising agent to treat cancers.
出处
《第三军医大学学报》
CSCD
北大核心
2000年第6期553-556,共4页
Journal of Third Military Medical University
关键词
苦参碱
肝细胞癌
细胞形态
HEPG2
细胞增殖
matrine
hepatocellular carcinoma
cell morphology
electron microscope
immunohistochemitry
