Study On Dendritic Cells Of Human Normal Endormetrium And Endometerioid Adenocarcinoma

Objective:1.To observe the morphological characteristics of dendritic cells ( DCs ) in human normal endometrium and endometrioid adenocarcinoma. 2. To study its the antigen presenting ability and apoptosis.3. To discuss the relationship between histologic grade, clinical stage and DC invasion in human endometrioid adenocarcinoma.Methods:20 cases of human normal endometrium in the postmenopausal tage and 35 cases of endometrioid adenocarcinoma were collected.1. All samples were respectively fixed in formalin solution and chilled in 2.5 % glutaraldehyde in 0.1M phosphate buffer and prepared for DC morphological study by immunohistochemistry and transmission election microscope.2. The expression of CD80 and CD86 in DC of all samples was evaluated via flow cytometry respectively. The expression of HLA, CD40 and CD54 in DC of all samples was respectively studied by double-label techniques and image analysis. 3. The apoptosis rate of DC was measured by double-label techniques and TUNEL technique. The expression of CD40, Fas and FasL in DC was respectively studied by double-label techniques and image analysis.4. DC in 35 cases of human endometrioid adenocarcinoma ( histologic grading: 14 well- differentiated cases, 12 mid-differentiated cases, 9 poorly-differentiated cases; clinical staging: 14 cases of stageⅠ, 13 cases of stageⅡ, 8 cases of stageⅢ～Ⅳ)were labeled by immunohistochemistry technique with S-100 protein. The volume density of the positive S-100 DC was evaluated by image analysis and stereology.Results:1. Lightmicroscopy and ultrastructure of dendritic cells of human normal endometriumSingle or numerous DCs were usually distributed in glands, vessels and glandular epithelial cells; but irregular, round or oval in shape. The cells were bigger in volume. Some cytoplasmic processes attached to each other. The nucleus was large in volume and irregular in shape.Under electron microscope, DCs were irregular in shape. Many cytoplasmic processes were thin and long, but, some processes were short and round. The vacuoles in the cytoplasm were somewhat round and numerous, but different in size. Some flocculent electron-densed granules were seen in the majority of vacuoles. Numerous round-shaped mitochondria appeared near the vacuoles. Rough endoplasmic reticulum (RER), the Golgi apparatus and lysosomes were rare. The nucleus was large in volume, round or oval in shape and located in a central position of the cells. Many condensed heterochromatin were seen under the nuclear membrane. Intercellular gap junction was visible.2. Lightmicroscopy and ultrastructure of dendritic cells of human endometrioid adenocarcinomaNumerous DCs were small in volume and round or regular in shape, but some were oval, spindly and polytangular. The cytoplasmic processes were invisible and the nucleus became obviously small.The cytoplasmic processes were obviously decreased in number. Mitochondria was relatively little in size. Round primary lysosome with high electron-densed granules and secondary lysosome with high or low electron-densed granules were seen frequently. DCs contained many rough endoplasmic reticulum (RER), the Golgi apparatus and ribosomes. The vacuoles with flocculent electron-densed granules were rare. Some special granules in cytoplasm were seen, whose surface like earphone were covered with a membrane. High electron-densed contents in the granules were near one side and the other side was bright. The nucleus became markedly small in volume, nephroid or hoofed in shape. The nucleus had little euchromatin and lots of heterochromatin under nuclear membrane.3. Expression of CDS0 and CD86 of dendritic cells in human normal endometrium and endometrioid adenocarcinomaThe expression of MHC-Ⅱ, CD80, CD86, CD40 and CD54 of DCs was considerablely lower than in human endormetrioid adenocarcinoma compared with that in normal endormetrium. (p＜0.05)4. Expression of HLA-DR, CD40 and CD54 of dendritic cells in human normal endometrium and endometrioid adenocarcinomaThe expression of HLA-DR, CD40 and CD54 of DCs in human normal endometrium was significant higher than that in endometrioid adenocarcinoma.(p＜0.05)5. Apoptosis of dendritic cells in human normal endometrium and endometrioid adenocarcinomaThe apoptosis rate of DCs in human normal endometrium was 6.82±0.53%, and was 15.95±0.87% in endometrioid adenocarcinoma. There was a significant difference between the apoptosis rate of DCs in in human normal endometrium and in endometrioid adenocarcinoma (p＜0.05).6. Expression of CD40, Fas/FasL(CD-95/CD-95L) and Bcl-2 of dendritic cells in human normal endometrium and endometrioid adenocareinomaThe expression of CD40, Fas(CD-95) and Bcl-2 of DCs in human normal endometrium was significantly higher than that in endometrioid adenocarcinoma (p＜0.05). Whereas, the intensity of FasL(CD-95L)of DCs was considerably lower than in human normal endormetrium compared with that in endormetrioid adenocarcinoma. (p＜0.05)7. Relationship between histologie grading, clinical staging and dendritic cells in human endometrioid adenoeareinomaIn human endometrioid adenocarcinoma, the volume density of DCs in well-differentiated group was statistically significantly higher than that in mid-differentiated group (p＜0.05). The same result was found as mid-differentiated group compared with poorly-differentiated group.In human endometrioid adenocarcinoma, the volume density of DCs in clinical stageⅠwas statistically significantly higher than that of clinical stageⅡ(p＜0.05). The same result was found as clinical stageⅡcompared with clinical stageⅢ～Ⅳ.Conclusion:1. In comparison to the DCs of normal human endormetrium, the DC of endormetrioid adenocarcinoma show morphological differences. Furthermore, these differences reflect the functional changes of the DCs in uptake, processing and presenting antigen.2. The considerable decrease of MHC-Ⅱ,CD80,CD86,CD40 and CD54 expression of DCs in human endormetrioid adenocarcinoma suggests that the antigen-presenting capability of the DCs of human endormetrioid adenocarcinoma is inhibited and may lead to tumor immune escape3. The apoptosis rate of DCs of human endormetrioid adenocarcinoma is higher than that of normal endormetrium. The lower levels of the CD40 andBcl-2 expression of DCs of human endormetrioid adenocarcinoma promote DC apoptosis, but, the lower expression of Fas and the higher expression of FasL of DCs of human endormetrioid adenocarcinoma indicate that the apoptosis of DCs might be regulated by a complicated mechanism.4. DCs invasion in human endormetrioid adenocarcinoma is correlated with histologic grade and clinical stage. The higher histologic grade and the earlier clinical stage is, the better are the prognosis in patients with endormetrioid adenocarcinoma.