Wang Jing, Deng Lu, Zhuang Huiyu, Liu Juanjuan, Liu Dawo, Li Xiao, Jin Shan, Zhu Liancheng, Wang Huimin, Lin Bei
1Department of Obstetrics and Gynaecology, Shengjing Hospital Affiliated To China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, 110004 Liaoning China.
Key Laboratory of Maternal-Fetal Medicine of Liaoning Province, Laboratory of Obstetrics and Gynecology of Higher Education of Liaoning Province, No. 7 Mulan Road, Xihu District, Benxi, 117000 Liaoning China.
Cancer Cell Int. 2019 Jun 13;19:161. doi: 10.1186/s12935-019-0864-4. eCollection 2019.
The interaction between human epididymis protein 4 (HE4) and annexin A2 (Annexin A2) has been found in ovarian cancer. However, it is dimness whether the interaction exists in other malignant tumors.
Real-time PCR, western blotting and immunocytochemistry were used to detect mRNA and proteins expression. Co-immunoprecipitation and double-labeling immunofluorescence were used to detect the interaction among HE4, ANXA2 and MMP2. MTS assay was used to test cell proliferation. Adhesion test was used to test cell adhesion. Flow cytometry was applied to examine cell cycle. The scratch test and Transwell assay was performed to detect the migration and invasion of various malignant cell lines.
Here we show that the overexpression of HE4 and ANXA2 in various malignant cells is a common phenomenon. HE4 and ANXA2 are co-localized in the cytoplasm and membrane of various tumor cells. ES-2 cells which had both high expression of HE4 and ANXA2 were much stronger in proliferation, adhesion, invasion, and migration than other tumor cells. HE4-ANXA2-MMP2 could form a triple protein complex. HE4 could mediate the expression of MMP2 via ANXA2 to promote cell migration progress.
The interaction of HE4 and ANXA2 exists in various types of cancer cells. HE4 and ANXA2 can promote the proliferation, adhesion, invasion, and migration of cancer cells. HE4-ANXA2-MMP2 form a protein complex and ANXA2 plays the role of "bridge". They performed together to promote cell migration.
已在卵巢癌中发现人附睾蛋白4（HE4）与膜联蛋白A2（Annexin A2）之间存在相互作用。然而，这种相互作用在其他恶性肿瘤中是否存在尚不清楚。
采用实时定量聚合酶链反应、蛋白质免疫印迹法和免疫细胞化学法检测mRNA和蛋白质表达。采用免疫共沉淀和双标记免疫荧光法检测HE4、ANXA2和基质金属蛋白酶2（MMP2）之间的相互作用。采用MTS法检测细胞增殖。采用黏附试验检测细胞黏附。应用流式细胞术检测细胞周期。进行划痕试验和Transwell试验以检测各种恶性细胞系的迁移和侵袭能力。
我们发现，HE4和ANXA2在各种恶性细胞中的过表达是一种常见现象。HE4和ANXA2共定位于各种肿瘤细胞的细胞质和细胞膜中。HE4和ANXA2均高表达的ES-2细胞在增殖、黏附、侵袭和迁移方面比其他肿瘤细胞更强。HE4-ANXA2-MMP2可形成三聚体蛋白复合物。HE4可通过ANXA2介导MMP2的表达，从而促进细胞迁移进程。
HE4与ANXA2的相互作用存在于各种类型的癌细胞中。HE4和ANXA2可促进癌细胞的增殖、黏附、侵袭和迁移。HE4-ANXA2-MMP2形成蛋白复合物，且ANXA2起到“桥梁”作用。它们共同发挥作用促进细胞迁移。
