ANXA7和HE4在子宫内膜癌中表达的实验研究

Experimental Study on Expression of ANXA7 and HE4 in Endometrial Carcinoma

目的探讨膜联蛋白A7(annexin A7,ANXA7)和人附睾蛋白4(human epididymis protein4,HE4)在子宫内膜癌中的表达及相关性。方法采用免疫组织化学方法检测石蜡包埋的子宫内膜组织中84例子宫内膜癌,30例子宫内膜不典型增生和18例正常子宫内膜组织中ANXA7和HE4蛋白的表达。采用Western Blot方法检测ANXA7和HE4的表达情况;通过免疫共沉淀技术和双重免疫荧光染色研究ANXA7和HE4之间的结构关系。结果免疫细胞化学检测的结果证实了ANXA7和HE4在子宫内膜癌细胞系中均有表达。免疫荧光显示通过红色荧光标记的ANXA7和通过绿色荧光标记的HE4在子宫内膜癌细胞系和不同子宫内膜组织的细胞膜和细胞质中的共表达,并且观察到重叠的橙色荧光。蛋白质印迹法结果证实ANXA7和HE4在子宫内膜癌细胞系中均有表达。免疫共沉淀测试检测到两种细胞系中ANXA7和HE4之间的相互作用。ANXA7在子宫内膜癌,恶性和非典型增生组织中的阳性表达率显著高于正常组织中的阳性表达率(P<0.05)。子宫内膜癌组HE4阳性表达率明显高于非典型增生组和正常对照组(P<0.05)。结论ANXA7和HE4在子宫内膜癌中均有表达。ANXA7和HE4的表达水平与子宫内膜癌的恶性生物学行为密切相关。

Objective To investigate the expression and correlation of ANXA7 and HE4 in endometrial carcinoma.Methods The expression of ANXA7 and HE4 protein in 84 cases of endometrial carcinoma,30 cases of endometrial atypical hyperplasia,and 18 cases of normal endometrial tissue samples was measured by using immunohistochemistry in paraffin embedded endometrial tissues.The expression of ANXA7 and HE4 was detected by Western Blot.The structural relationship between ANXA7 and HE4 was explored by immunoprecipitation and double immunofluorescent staining.Results The results of immunohistochemistry confirmed that ANXA7 and HE4 were expressed in endometrial cancer cell lines.Immunofluorescence showed co-expression of red fluorescently labeled ANXA7 and green fluorescently labeled HE4 in the cell membrane and cytoplasm of endometrial cancer cell lines and different endometrial tissues,and overlapping orange fluorescence was observed.Western blotting results confirmed that ANXA7 and HE4 were expressed in endometrial cancer cell lines.The co-immunoprecipitation results showed the interaction between ANXA7 and HE4 in both cell lines.The positive expression rate of ANXA7 in endometrial cancer,malignant and atypical hyperplasia tissues was significantly higher than that in normal tissues(P<0.05).The positive expression rate of HE4 in the endometrial cancer group was significantly higher than that in the atypical hyperplasia group and the normal control group(P<0.05).Conclusion ANXA7 and HE4 are expressed in endometrial carcinoma,and the expression levels of ANXA7 and HE4 are closely related to the malignant biological behavior of endometrial carcinoma.