Shen Dao-Fu, Liu Xin, Yang Xue-Feng, Fang Lei, Gao Yang, Zhao Shuang, Wu Ji-Cheng, Shi Shuai, Li Jun-Jun, Zhao Xiang-Xuan, Gou Wen-Feng, Zheng Hua-Chuan
Cancer Center, Key Laboratory of Brain and Spinal Cord Injury of Liaoning Province, and Laboratory Animal Center, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, 121001, China.
Department of Clinical Laboratory, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, 121001, China.
Tumour Biol. 2016 Mar;37(3):2909-24. doi: 10.1007/s13277-015-4103-x. Epub 2015 Sep 26.
Parafibromin is a protein encoded by hyperparathyroidism 2 (HRPT2) and its downregulated expression is involved in the pathogenesis of parathyroid, breast, gastric, colorectal, lung, head and neck cancers. We aimed to investigate the roles of parafibromin expression in tumorigenesis, progression, or prognostic evaluation of ovarian cancers. HRPT2-expressing plasmid was transfected into ovarian cancer cells with the phenotypes and related molecules examined. The messenger RNA (mRNA) and protein expression of parafibromin were also examined in ovarian normal tissue, benign and borderline tumors and cancers by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, or immunohistochemistry respectively. It was found that parafibromin overexpression caused a lower growth, migration and invasion, higher sensitivity to cisplatin and apoptosis than the mock and control (P < 0.05). The transfectants showed the hypoexpression of phosphoinositide 3-kinase (PI3K), Akt, p70 ribosomal protein S6 kinase (p70s6k), Wnt5a, B cell lymphoma-extra large (Bcl-xL), survivin, vascular endothelial growth factor (VEGF) and matrix metallopeptidase 9 (MMP-9) than the mock and control at both mRNA and protein levels (P < 0.05). According to real-time PCR, parafibromin mRNA level was lower in ovarian benign tumors and cancers than normal ovary (P < 0.05), while parafibromin was strongly expressed in metastatic cancers in omentum than primary cancers by Western blot. Immunohistochemically, parafibromin expression was stronger in primary cancers than that in ovarian normal tissue (P < 0.05) but weaker than the metastatic cancers (P < 0.05) with a positive correlation with dedifferentiation, ki-67 expression and the lower cumulative survival rate (P < 0.05). These findings indicate that parafibromin downregulation might promote the pathogenesis, dedifferentiation and metastasis of ovarian cancers possibly by suppressing aggressive phenotypes, such as proliferation, cell cycle, apoptosis, migration and invasion.
副纤维瘤蛋白是由甲状旁腺功能亢进2（HRPT2）编码的一种蛋白质，其表达下调与甲状旁腺癌、乳腺癌、胃癌、结直肠癌、肺癌、头颈癌的发病机制有关。我们旨在研究副纤维瘤蛋白表达在卵巢癌发生、发展或预后评估中的作用。将表达HRPT2的质粒转染到卵巢癌细胞中，并检测其表型和相关分子。分别通过逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法或免疫组织化学检测卵巢正常组织、良性和交界性肿瘤及癌组织中副纤维瘤蛋白的信使核糖核酸（mRNA）和蛋白质表达。结果发现，与空载体和对照组相比，副纤维瘤蛋白过表达导致细胞生长、迁移和侵袭能力降低，对顺铂的敏感性增加以及凋亡增加（P < 0.05）。转染细胞在mRNA和蛋白质水平上均显示磷酸肌醇3激酶（PI3K）、蛋白激酶B（Akt）、p70核糖体蛋白S6激酶（p70s6k）、Wnt5a、B细胞淋巴瘤-超大蛋白（Bcl-xL）、生存素、血管内皮生长因子（VEGF）和基质金属蛋白酶9（MMP-9）的表达低于空载体和对照组（P < 0.05）。实时定量PCR结果显示，卵巢良性肿瘤和癌组织中副纤维瘤蛋白mRNA水平低于正常卵巢组织（P < 0.05），而蛋白质免疫印迹法显示大网膜转移癌中副纤维瘤蛋白的表达强于原发性癌。免疫组织化学结果显示，原发性癌中副纤维瘤蛋白的表达强于卵巢正常组织（P < 0.05），但弱于转移癌（P < 0.05），且与去分化、ki-67表达及较低的累积生存率呈正相关（P < 0.05）。这些发现表明，副纤维瘤蛋白表达下调可能通过抑制增殖、细胞周期、凋亡、迁移和侵袭等侵袭性表型来促进卵巢癌的发病机制、去分化和转移。
