Wang Fei, Fang Meixia, Kong Min, Wang Changhe, Xu Yuting
Department of Gynecology, Jining No.1 People's Hospital, Jining, Shandong, China.
J Biochem Mol Toxicol. 2020 Sep;34(9):e22525. doi: 10.1002/jbt.22525. Epub 2020 May 5.
The influences of Vinculin on many cancers were blurry, including ovarian cancer. Thus, we concentrated on the efficient role of Vinculin in ovarian cancer and explored the potential mechanism(s). Expression of Vinculin in ovarian cancer tissues and cell lines was investigated by real-time polymerase chain reaction, immunohistochemistry, and Western blot. The Kaplan-Meier manner with the logrank was performed to assess overall survival. We further evaluated the relations between Vinculin expression and clinicopathological features of ovarian cancer. Moreover, Vinculin was overexpressed or silenced by respectively transfection with pcDNA-Vinculin or small interfering (si-Vinculin) into human ovarian cancer cell line Caov3 or human ovarian epithelial cell line (HOEpiC). Thereafter, cell viability, cell apoptosis, and migration were checked by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, flow cytometer, and scratch assay, respectively. Likewise, the apoptosis- and migration-related proteins were distinguished by Western blot. Compared to the nontumor tissues or HOEpiC cells, Vinculin was significantly lower expressed in the ovarian cancer tissues and cells. Furthermore, we found out that Vinculin was primarily distributed at the cell membrane and cytoplasm. Moreover, Vinculin was negatively associated with International Federation of Gynecology and Obstetrics stage, grade, and distant metastasis. Overexpression of Vinculin dramatically weakened cell viability and migration and stimulated apoptosis. Conversely, suppression of Vinculin showed opposite results. Vinculin presents unfavorable prediction in ovarian cancer and inhibits ovarian cancer proliferation and migration.
纽蛋白（Vinculin）对包括卵巢癌在内的多种癌症的影响尚不明确。因此，我们聚焦于纽蛋白在卵巢癌中的作用，并探究其潜在机制。通过实时聚合酶链反应、免疫组织化学和蛋白质免疫印迹法研究了纽蛋白在卵巢癌组织和细胞系中的表达情况。采用Kaplan-Meier法和对数秩检验评估总生存期。我们进一步评估了纽蛋白表达与卵巢癌临床病理特征之间的关系。此外，通过分别将pcDNA-纽蛋白或小干扰RNA（si-纽蛋白）转染至人卵巢癌细胞系Caov3或人卵巢上皮细胞系（HOEpiC）中，使纽蛋白过表达或沉默。此后，分别通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐法、流式细胞仪和划痕试验检测细胞活力、细胞凋亡和迁移情况。同样，通过蛋白质免疫印迹法鉴定凋亡和迁移相关蛋白。与非肿瘤组织或HOEpiC细胞相比，纽蛋白在卵巢癌组织和细胞中的表达显著降低。此外，我们发现纽蛋白主要分布于细胞膜和细胞质中。而且，纽蛋白与国际妇产科联盟分期、分级及远处转移呈负相关。纽蛋白过表达显著削弱细胞活力和迁移能力，并促进细胞凋亡。相反，抑制纽蛋白表达则产生相反的结果。纽蛋白在卵巢癌中呈现不良预后，并抑制卵巢癌的增殖和迁移。
