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目的:探讨雌激素受体(estrogen receptor,ER)阳性的乳腺癌患者癌组织中Efp和Plk3的表达及其临床意义和可能的作用机制。方法:采用免疫组织化学法检测于2010年1月至6月在河北医科大学第四医院乳腺科住院的86例ER阳性患者乳腺癌组织中Efp和Plk3的蛋白表达,并分析两者表达的相关性及与临床病理指标的关系。采用qRT-PCR法检测乳腺癌细胞MCF-7和MDA-MB-231中ER、Efp mRNA的表达情况,采用RT-PCR及Western blotting法检测雌激素刺激后ER阳性乳腺癌细胞MCF-7中Efp、Plk3基因的表达变化,采用Western blotting法检测雌激素及蛋白酶抑制剂MG132处理后MCF-7细胞中Efp、Plk3的表达变化。结果:86例ER阳性的乳腺癌组织中,55例Efp表达阳性(64.0%),28例Plk3表达阳性(32.6%)。Efp表达阳性组织中Plk3表达阳性为23.6%,Efp表达阴性组织中Plk3表达阳性为48.4%,Efp和Plk3蛋白表达存在明显的负相关(P<0.05),Efp蛋白表达与乳腺患者的淋巴结转移状况呈明显正相关(P<0.05),Plk3蛋白表达与乳腺癌患者的淋巴结转移状况呈明显负相关(P<0.05)。MCF-7细胞ER mRNA高表达,而MDA-MB-231细胞的ER mRNA低表达。MDA-MB-231细胞经雌激素刺激后,Efp mRNA的表达未见明显改变。MCF-7经雌激素刺激后Efp mRNA及蛋白的表达显著增加(P<0.05),而Plk3 mRNA的表达未见明显改变,未检测到Plk3蛋白的表达。MG132处理后MCF-7细胞中Plk3的蛋白表达明显上调,MCF-7经雌激素刺激后,再用MG132处理,Efp的蛋白表达显著增加(P<0.05),而Plk3蛋白表达明显下降(P<0.05)。结论:ER阳性乳腺癌中Efp和Plk3蛋白表达存在明显的负相关,Efp可促进Plk3的蛋白降解,并可能参与了ER阳性乳腺癌患者内分泌治疗的耐药过程。
Objective: To investigate the expression of Efp and Plk3 in cancerous tissues of patients with estrogen receptor (ER) positive breast cancer and its clinical significance and possible mechanism. Methods: Immunohistochemical method was used to detect the protein expression of Efp and Plk3 in 86 cases of ER-positive breast cancer patients hospitalized in Department of Breast Medicine, the Fourth Hospital of Hebei Medical University from January to June in 2010, and to analyze the correlation between them Sex and its relationship with clinicopathological parameters. The expression of ER and Efp mRNA in breast cancer cells MCF-7 and MDA-MB-231 was detected by qRT-PCR. The expression of Efp in ER-positive breast cancer cells MCF-7 was detected by RT-PCR and Western blotting , Plk3 gene expression change, using Western blotting assay estrogen and protease inhibitor MG132 treatment MCF-7 cells Efp, Plk3 expression changes. Results: Of 86 ER-positive breast cancers, 55 were positive for Efp (64.0%) and 28 were positive for Plk3 (32.6%). The positive expression of Plk3 in Efp positive tissues was 23.6%, Plk3 positive in Efp negative tissues was 48.4%, there was a significant negative correlation between Efp and Plk3 protein expression (P <0.05). Efp protein expression was correlated with lymph node metastasis (P <0.05). Plk3 protein expression was negatively correlated with lymph node metastasis in breast cancer patients (P <0.05). ER mRNA was highly expressed in MCF-7 cells, whereas ER mRNA in MDA-MB-231 cells was low. The expression of Efp mRNA in MDA-MB-231 cells stimulated by estrogen showed no significant change. The mRNA and protein expression of Efp in MCF-7 stimulated by estrogen increased significantly (P <0.05), while the expression of Plk3 mRNA did not change significantly. The expression of Plk3 protein was not detected. The protein expression of Plk3 in MCF-7 cells was significantly up-regulated after treatment with MG132. The protein expression of Efp was significantly increased (P <0.05) and the expression of Plk3 protein was significantly decreased after MCF-7 stimulation with estrogen and MG132 (P < 0.05). Conclusion: There is a significant negative correlation between Efp and Plk3 protein expression in ER positive breast cancer. Efp may promote the protein degradation of Plk3 and may be involved in the endocrine resistance process in patients with ER positive breast cancer.