【摘要】 目的探讨卵巢癌组织中Hippo-Yes相关蛋白(Yes-associated protein,YAP)的表达与临床病理参数的关系,及其对卵巢癌细胞增殖和凋亡的影响。方法免疫组化染色技术检测湖南省肿瘤医院病理科2010-01-01-2013-12-31保存的68例卵巢癌及38例良性卵巢组织中YAP蛋白表达,分析YAP表达与卵巢癌临床病理参数的相关性;逆转录病毒介导的YAP shRNA干扰人卵巢癌EFO-21细胞中YAP表达,蛋白质印迹法检测干扰效率,5-溴脱氧尿嘧啶核苷(5-Bromo-2-deoxy Uridine,BrdU)细胞增殖分析和流式细胞术检测肿瘤细胞增殖和凋亡变化。结果卵巢癌组织YAP蛋白阳性表达率为61.8%(42/68),良性卵巢组织为10.5%(4/38),差异有统计学意义,χ2=26.054,P<0.001。卵巢癌组织中YAP蛋白阳性表达率在高临床分期(χ2=8.600,P=0.003)和高病理分级(χ2=5.688,P=0.017)中显著升高。YAP shRNA显著降低卵巢癌EFO-21细胞中YAP蛋白表达,对照组YAP蛋白表达为0.96±0.16,实验组为0.14±0.04,P<0.001。YAP shRNA导致肿瘤细胞增殖能力显著减弱,对照组细胞增殖水平为121.00±3.19,实验组为55.83±3.56,P<0.001。YAP shRNA并诱导细胞凋亡,对照组凋亡率为(8.75±0.15)%,实验组为(32.75±4.76)%,P<0.001。结论 YAP蛋白在卵巢癌组织中表达异常升高并核内蓄积,其阳性表达与高病理分级和高临床分期显著有关,干扰YAP表达导致肿瘤细胞增殖减弱和凋亡增加,提示YAP通过促进肿瘤细胞生长发挥促癌作用,其可能成为卵巢癌靶向治疗的潜在靶点。更多还原

【Abstract】 OBJECTIVE To investigate the expression of Yes-associated protein(YAP)in ovarian cancer tissues and its effects on cell proliferation and apoptosis in ovarian cancer cells.METHODS We detected YAP expression in68 samples of paraffin-embedded ovarian cancer and 38 samples of normal ovarian tissues,which were stored in Department of Pathology,Tumor Hospital of Hunan Province during 1st Jan.2010 to 31th Dec.2013,using immunohistochemistry and analyzed the clinicalpathological significance of YAP in ovarian carcinoma.Silence of YAP by retrovirus-mediated YAP shRNA in EFO-21 cells was confirmed by Western blot.The cell proliferation and apoptosis were analyzed using5-Bromo-2-deoxy Uridine(BrdU)cell proliferation assay and flow cytometry.RESULTS The positive expression rates of YAP in ovarian cancer tissues and normal ovarian tissues were 61.8%(42/68)and 10.5%(4/38),respectively.The difference was statistically significant(χ2=26.054,P<0.001).The positive expression rate of YAP was significantly higher in advanced clinical stage(χ2=8.600,P=0.003)and high histological grading(χ2=5.688,P=0.017).Specific shRNA significantly decreased the protein level of YAP in EFO-21cells(control group:0.96±0.16 vs experimental group:0.14±0.04,P<0.001),YAP knockdown decreased cell proliferation(control group:121.00±3.19 vs experimental group:55.83±3.56,P<0.001)and induced cell apoptosis [control group:(8.75±0.15)% vs experimental group:(32.75±4.76)%,P<0.001]in EFO-21 cells.CONCLUSIONS The elevated expression and nuclear accumulation of YAP is observed in ovarian cancer tissues,its high-expression is associated with high histological grading and advanced clinical stage.YAP knockdown lead to decreased proliferation and increased apoptosis in EFO-21 cells,suggesting that YAP acts as an oncogene by promoting tumor cell growth and may be a potential therapeutic target for ovarian cancer.更多还原