Liu Qicai, Li Ermao, Huang Long, Cheng Minsheng, Li Li
Guangdong Provincial Biomedical Engineering Technology Research Center for Cardiovascular Disease; Sino-Japanese Cooperation Platform for Translational Research in Heart Failure; Laboratory of Heart Center, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China.
Department of Cardiology; Heart Center, Zhujiang Hospital, Southern Medical University.
J Cancer. 2018 Jan 1;9(2):424-432. doi: 10.7150/jca.21375. eCollection 2018.
The limb-bud and heart gene () was discovered in the early 21st century and is specifically expressed in the mouse embryonic limb and heart development. Increasing evidences have indicated that LBH not only plays an important role in embryo development, it is also closely correlated with the occurance and progression of many tumors. However, its function in prostate cancer (PCa) is still not well understood. Here, we explored the effects of LBH on the proliferation and migration of the PCa cell line PC3M. LBH expression in tissues and cell lines of PCa was detected by immunohistochemistry and Western blotting. Lentivirus was used to transduct the LBH gene into the PC3M cells. Stable LBH-overexpressing PC3M-LBH cells and PC3M-NC control cells were obtained via puromycin screening. Cell proliferation was examined using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cell cycle distribution and apoptosis rate were investigated using flow cytometry. Cell migration was studied using the Transwell assay. LBH expression level was down-regulated in 3 different PCa cell lines, especially in PC3M cells, compared with the normal prostate epithelial cells(RWPE-1). Cell lines of LBH-upregulated PC3M-LBH and PC3M-NC control were successfully constructed. Significantly increased LBH expression level and decreased cyclin D1 and cyclin E2 expression level was found in PC3M-LBH cells as compared to the PC3M-NC cells. The overexpression of LBH significantly inhibited PC3M cell proliferation in vitro and tumor growth in nude mice. LBH overexpression in PC3M cell, also induced cell cycle G0/G1 phase arrest and decreased the migration of PC3M cells. : Our results reveal that LBH expression is down-regulated in the tissue and cell lines of PCa. LBH overexpression inhibits PC3M cell proliferation and tumor growth by inducing cell cycle arrest through down-regulating cyclin D1and cyclin E2 expression. LBH might be a therapeutic target and potential diagnostic marker in PCa.
肢体芽和心脏基因（LBH）于21世纪初被发现，在小鼠胚胎肢体和心脏发育中特异性表达。越来越多的证据表明，LBH不仅在胚胎发育中起重要作用，还与许多肿瘤的发生和进展密切相关。然而，其在前列腺癌（PCa）中的功能仍未完全清楚。在此，我们探讨了LBH对前列腺癌细胞系PC3M增殖和迁移的影响。通过免疫组织化学和蛋白质印迹法检测PCa组织和细胞系中的LBH表达。使用慢病毒将LBH基因转导至PC3M细胞中。通过嘌呤霉素筛选获得稳定过表达LBH的PC3M-LBH细胞和PC3M-NC对照细胞。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法检测细胞增殖。使用流式细胞术研究细胞周期分布和凋亡率。使用Transwell法研究细胞迁移。与正常前列腺上皮细胞（RWPE-1）相比，3种不同的PCa细胞系中LBH表达水平下调，尤其是在PC3M细胞中。成功构建了LBH上调的PC3M-LBH和PC3M-NC对照细胞系。与PC3M-NC细胞相比，PC3M-LBH细胞中LBH表达水平显著升高，细胞周期蛋白D1和细胞周期蛋白E2表达水平降低。LBH的过表达显著抑制PC3M细胞的体外增殖和裸鼠体内肿瘤生长。PC3M细胞中LBH的过表达还诱导细胞周期G0/G1期阻滞并降低PC3M细胞的迁移。我们的结果表明，PCa组织和细胞系中LBH表达下调。LBH的过表达通过下调细胞周期蛋白D1和细胞周期蛋白E2的表达诱导细胞周期阻滞，从而抑制PC3M细胞增殖和肿瘤生长。LBH可能是PCa的治疗靶点和潜在诊断标志物。
