【作者】 沈培；

【导师】 李继承；

【作者基本信息】 浙江大学 ， 细胞生物学， 2009， 硕士

【摘要】 研究背景:间皮细胞覆盖在浆膜腔及腔内脏器的表面,具有屏障、免疫、物质转运等功能。淋巴孔是毛细淋巴管在浆膜间皮细胞之间的微小开口,通过淋巴孔,浆膜腔与淋巴管系直接相通。淋巴孔具有物质吸收、免疫等功能。肿瘤腹膜转移一般经历细胞游离、粘附、侵袭、生长四个阶段。在肿瘤腹膜转移时,间皮细胞和淋巴孔形态会发生改变。研究目的:研究腹腔接种肿瘤细胞后,膈腹膜、肠系膜上的间皮细胞及淋巴孔在肿瘤腹膜转移过程中不同时期的变化,探讨间皮细胞及淋巴孔在肿瘤腹膜转移过程中发挥的作用。研究方法:小鼠腹腔接种HeA-H细胞后,每天记录体重增长状况,以确定收集腹水的时间点。接种后每两天取一次膈腹膜及肠系膜,扫描电镜观察腹腔接种肿瘤细胞后膈腹膜和肠系膜上间皮细胞及淋巴孔在不同时期的变化。扫描电镜观察用腹水连续处理后膈腹膜和肠系膜间皮细胞及淋巴孔的变化。以Boyden小室法研究HepA-H细胞侵袭能力。以ELISA法测定腹水中VEGF含量。肠系膜消化法培养间皮细胞,建立间皮细胞体外培养体系,为肿瘤转移过程中间皮细胞和淋巴孔的体外研究建立基础。研究结果:小鼠腹腔接种HepA-H肿瘤细胞后,体重较对照组有明显增长,第11天后体重增长减缓,第14天后出现死亡。研究发现肿瘤腹膜转移过程中间皮细胞发生明显肿胀,淋巴孔开放数目增加,孔中可见肿瘤细胞。腹水连续处理后,较之对照组,膈腹膜及肠系膜上淋巴孔开放明显增加,膈腹膜间皮细胞肿胀,绒毛增生,肠系膜间皮细胞边界出现明显微绒毛。Boyden小室测定HepA-H细胞侵袭能力为23.4±1.5%。腹水中VEGF含量为5.43±0.23ng/ml。从细胞形态及免疫荧光两方面的检测,确定肠系膜消化法培养之细胞为间皮细胞。结论:膈腹膜间皮细胞在HepA-H肝癌细胞腹膜转移过程中,细胞体肿胀,间皮细胞间隙增大,微绒毛增生,肿瘤细胞种植于间皮细胞之间。肠系膜间皮细胞形态变化不大,后期可见结节状肿瘤细胞。肿瘤细胞可通过淋巴孔传播。更多还原

【Abstract】 Background:Mesothelial cells cover the entire surface of the three serosal cavities and the internal organs.The mesothelial cells have many functions including protection,immune response,transportation and so on.Lymphatic stomata are small openings of lymphatic capillary on the free surface of mesothelium.The peritoneal cavity and pericardial cavity are connected with lymphatic system via these small openings which have the function of active absorption.Dissemination of tumors requires four steps:detachment of cancer cells from the primary tumor,attachment to distant peritoneum,invasion into subperitoneal space,and proliferation.Changes of mesothelial cells happen during the dissemination of tumor cells.Objective:To study the morphological changes of peritoneal mesothelial cells and lymphatic stomata on the diaphragm and mesentery during the dissemination of HepA-H cells.Methods:After injection of HepA-H ascites tumor to the peritoneal cavity, diaphragm and mesentery were observed by SEM at different time point.The fluid of ascites tumor was injected to the peritoneal cavity for 3 days.Then the two tissues were examined by SEM.Migration ability of HepA-H cells were tested by Boyden chamber.VEGF level was assayed with ELISA kit.Mesothelial cells from mesentery were cultured in vitro.Results:Compared to the control,the weight of mice showed an obvious growth. The growth speed decreased after the 11^th day since injection of HepA-H.Death occurred in the 14^th day.Morphological changes was observed during dissemination of ascites tumor.The stomata showed larger in size and quantity both on diaphragm and on mesentery.Tumor cells were also seen in the stomata.The migration ability tested by Boyden chamber was 23.4±1.5%.VEGF level in ascites was 5.43±0.23ng/ml.The morphological observation and immunofluorescence staining tests proved the cells collected from mesentery were mesothelial cells.Conclusion:Mesothelial cells on diaphragm showed tumid cell bodies,enlarged spaces between mesothelial cells,and increased microvilli during metastasis of tumor cells.Tumor cells were also observed planting between the mesothelial cells.Little morphological change were shown in mesenteric mesothelial cell.Nodular tumor cells can be seen in the late.Tumor cells were also seen getting through the lymphatic stomata.更多还原