25-甲氧基-达玛烷-3β、12β、20-三醇和青蒿素通过下调睾丸特异性蛋白酶50 (TSP50)表达协同抑制MDA-MB-231细胞增殖

2016年4月，东北师范大学可用药基因与蛋白筛选国家工程实验室；东北师范大学农业与医药基因工程教育部研究中心；沈阳药科大学食品科学工程学院；东北师范大学遗传与细胞学研究所(National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun 130024, China；Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University,Changchun 130024, China；Food Science Engineering, Shenyang Pharmaceutical University,Shenyang 110016, China；Institute of Genetics and Cytology, Northeast Normal University,Changchun 130024, China) Danfeng Wang老师研究团队在《Tumor Biology》上发表论文：

“25-methoxyl-dammarane-3β, 12β, 20-triol and artemisinin synergistically inhibit MDA-MB-231 cell proliferation through downregulation of testes-specific protease 50 (TSP50) expression”

“25-甲氧基-达玛烷-3β、12β、20-三醇和青蒿素通过下调睾丸特异性蛋白酶50 (TSP50)表达协同抑制MDA-MB-231细胞增殖”

Abstract：

Purpose: To explore the biological functions and mechanism of Opa interacting protein 5 (OIP5) in bladder cancer (BC).

Methods: We investigated the expression of OIP5 in BC through immunohistochemical staining (IHC) and its correlation with clinicopathologic features of BC patients. Moreover, knockdown of OIP5 was performed in BC cell lines and colony formation capacity, cell growth curve, cell cycle phase and cell apoptosis assay was applied for investigating the roles of OIP5 in BC. Moreover, the expression of OIP5 was validated through the Cancer Genome Atlas (TCGA) database. The diagnosis value of OIP5 was accessed by receiver operating characteristic (ROC) analysis in TCGA database.

Results: The expression of OIP5 in BC tissues was significantly higher than that in adjacent non-tumor tissues and bladder mucosa tissues with chronic cystitis. Higher protein expression level of OIP5 predicted shorter survival time in patients with BC, which was significantly correlated with larger tumor size, high-grade tumor and advanced T classification. The expression of OIP5 was considerably decreased after lentivirus infection both at mRNA and protein levels. Functional assay displayed that silencing of OIP5 inhibited colony formation capacity and cell growth in BC cell lines. Cell cycle assays indicated that suppressed OIP5 disturbed the balance of the cell cycle in BC cell lines, which increased the cell population of the G1 phase and decreased the cell population of the S phase. Furthermore, knockdown of OIP5 expression enhanced cell apoptosis process. The expression of OIP5 was significantly up-regulated in BC compared with adjacent non-tumor tissues based on TCGA database. OIP5 had the potential diagnostic value for BC.

Conclusions: Our work demonstrated that OIP5 might function as an oncogene to promote colony formation capacity and cell growth, arrest cell cycle and suppress cell apoptosis in bladder cancer.

摘要：

目的:探讨Opa相互作用蛋白5 (Opa interacting protein 5, OIP5)在膀胱癌(BC)中的生物学功能及机制。

方法:通过免疫组化染色(IHC)研究OIP5在BC中的表达及其与BC患者临床病理特征的相关性。在BC细胞株上敲低OIP5，通过集落形成能力、细胞生长曲线、细胞周期和细胞凋亡等实验研究OIP5在BC中的作用。此外，通过癌症基因组图谱(TCGA)数据库验证OIP5的表达。通过TCGA数据库的受试者工作特征(ROC)分析获得OIP5的诊断价值。

结果:慢性膀胱炎患者BC组织中OIP5的表达明显高于癌旁非肿瘤组织和膀胱黏膜组织。OIP5蛋白表达水平越高，BC患者生存时间越短，且与肿瘤大小较大、肿瘤级别高、T分期高相关。慢病毒感染后，OIP5在mRNA和蛋白水平上的表达均显著降低。功能实验表明，OIP5的沉默抑制了BC细胞系的集落形成能力和细胞生长。细胞周期实验表明，抑制OIP5扰乱了BC细胞系细胞周期的平衡，使G1期细胞数量增加，S期细胞数量减少。此外，OIP5表达下调可促进细胞凋亡过程。TCGA数据库显示，与癌旁非肿瘤组织相比，BC组织中OIP5的表达明显上调。OIP5对BC有潜在的诊断价值。

结论:OIP5可能作为癌基因在膀胱癌中具有促进细胞集落形成能力和细胞生长、阻滞细胞周期和抑制细胞凋亡的作用。

该论文中，人膀胱癌T24细胞(BC-T24)、人胚胎肾(HEK) 293T细胞和人膀胱癌5637细胞(BC-5637)的体外培养是使用Ausbian特级胎牛血清完成的。欲了解或购买Ausbian特级胎牛血清可以联系北京缔一生物400-166-8600.