【作者】 桑娜；

【导师】 李景和；

【作者基本信息】 中南大学 ， 病理学与病理生理学， 2010， 硕士

【摘要】 第一部分EGFL7在胃癌组织中的表达及其临床病理关系的研究目的：探讨胃癌组织中表皮生长因子样结构域7(EGFL7)的表达及其与胃癌临床病理特征的关系。方法：采用免疫组织化学SP法分别检测45例胃癌组织中EGFL7的表达水平及微血管密度(MVD,CD34标记),分析EGFL7的表达与MVD及胃癌临床病理特征之间的关系。结果：EGFL7的表达在有淋巴结转移组中的阳性率(84.6%)高于无淋巴结转移组(57.9%),在浸润浆膜层组中的阳性率(84.4%)高于未浸润浆膜层组(46.2%)。胃癌组织中EGFL7的表达与淋巴结转移和浸润程度呈正相关(p<0.05),但与患者的年龄、性别、肿瘤的分化程度无关(p>0.05)。胃腺癌低分化组MVD(37.62±10.42)高于高分化组(27.91±9.93)和中分化组(28.40±9.18)；浸润至浆膜层组MVD(34.25±10.43)高于未至浆膜层组(26.69±8.66)；有淋巴结转移组MVD(33.00±9.99)高于无淋巴结转移组(25.94±9.34)。癌组织中MVD值与分化程度、浸润深度及有无淋巴结转移呈正相关(p<0.05),但与年龄、性别无关(p>0.05)。在EGFL7表达阳性的肿瘤组织中MVD均值为33.80±10.56,高于EGFL7表达阴性的肿瘤组织中MVD均值26.00±7.21(p<0.05),采用Pearson积差相关系数进行分析,发现MVD值与EGFL7的表达成正相关(r=0.313,p<0.05)。结论：EGFL7的高表达可能促进了胃癌的浸润和转移,其机制可能与促进胃癌组织中的血管新生有关。第二部分shRNA介导的瘤源性EGFL7基因沉默对人脐静脉内皮细胞生物学行为的影响目的：探讨瘤源性EGFL7对人脐静脉内皮细胞(HUVEC)生物学行为的影响。方法：利用免疫细胞化学、western bolt方法检测四株胃癌细胞SGC7901、BGC823、MKN28、MKN45中EGFL7的表达,选取表达最强的一株用于后续实验；设计合成针对EGFL7基因的RNA干扰序列,建立稳定转染的EGFL7干扰胃癌细胞株,将该株细胞与HUVEC体外共培养,检测HUVEC增殖、粘附及迁徙能力的改变。结果：1.SGC7901、BGC823、MKN45和MKN28四株胃癌细胞均表达EGFL7蛋白,且表达均定位于细胞浆中；western blot检测显示EGFL7在BGC823中的表达最强；2.将稳定转染EGFL7基因RNA干扰序列的BGC823细胞株(命名为BGC2-13)、未转染BGC823细胞及转染阴性质粒的BGC823细胞株(命名为BGCHK)与HUVEC共培养,发现降低瘤源性EGFL7的分泌导致HUVEC增殖速率降低,同时可使其粘附能力、迁徙能力下降(p<0.05)。结论：瘤源性EGFL7能够促进内皮细胞的增殖、粘附及迁徙,从而有可能促进血管的新生。更多还原

【Abstract】 Party I The study on clinical significance of the expression of EGFL7 in gastric carcinomaObjective:To explore the expression of EGFL7in gastric carcinoma and its correlation with tumor clinical and pathological features.Methods:The expression of EGFL7 protein and MVD in 45 cases gastric carcinoma were detected by immunohistochemical methods.Result:The expression of EGFL7in the team which has lymph node metastasis (84.6%) is higher than the team which hasn’t (57.9%),in the serosal invasion team (84.4%) is higher than the team which doesn’t have serosal invasion (46.2%). The expression of EGFL7 in gastric carcinoma is related with tumor invasion and lymph node metastasis (p<0.05),but isn’t related with the patients’age, sex and tumor differentiation (p>0.05)The MVD of poorly differentiation team (37.62±10.42)is higher than the well-differentiation team (27.91±9.93) and differentiated team (28.4±9.18). The serosal invasion team’s MVD(34.25±10.43)is higher than the team which is not (26.69±8.66),the team’s MVD which has lymph node metastasis (33.00±9.99) is higher than the team which hasn’t (25.94±9.34).The MVD is related with tumor differentiation, invasion and lymph node metastasis, but isn’t related with the age and sex (p>0.05)MVD in the tumor which express EGFL7 is 33.80±10.56, that is higher than the MVD in the tumor which don’t express EGFL7 (26.00±7.21) (p<0.05).Pearson analysis shows MVD and EGFL7 expression is positive correlation (r=0.313,p<0.05)Conclusion:High expression of EGFL7 would progress the infiltration and metastasis of gastric caicinoma. The mechanism maybe related with angiogenesis. PartyⅡStudy on the effect on human umbilical vein endothelial cell biology behavior when EGFL7 gene silence in gastric carcinoma cellObjective:To explore the effect on HUVEC’s biological behavior when silence EGFL7 in gastric carcinoma cell.Methods:Use immunocytochemistry and western blot to defect EGFL7 expression in 4 gastric carcinoma cell line, SGC7901, BGC823, MKN28 and MKN45, select the highest expression cell line for next experiment. Design and synthesis RNA interference sequence for EGFL7, establish a stable transfected gastric carcinoma cell line, use this cell line and HUVEC do co-culture in vitro, detect the change in HUVEC’s proliferation, adhesion and migration ability.Result:1.The 4 gastric carcinoma cell line SGC7901, BGC823, MKN45, MKN28 all express EGFL7 protein and the protein locate in the cytoplasm. Western blot show the BGC823 cell line has the highest expression; 2.The BGC823 cell line which is stable transfected (named BGC2-13), BGC823 cell line which is untranfected and BGC823 cell line which is transfected negative plasmid(named BGCHK)do co-culture with HUVEC, we find that lower EGFL7 expression cause the proliferation rate of HUVEC lower, and lower its adhesion and migration ability also (p<0.05)Conclusion:Tumor-derived can progress HUVEC’s proliferation, adhesion and migration ability, and then would progress angiogenesis.更多还原