Liu Ying, Zheng Dongping, Liu Mingming, Bai Jiao, Zhou Xi, Gong Baolan, Lü Jieyu, Zhang Yi, Huang Hui, Luo Wenying, Huang Guangrong
Department of Obstetrics and Gynecology, Renmin Hospital, Hubei University of Medicine, Shiyan, 442000, Hubei Province, China.
Ultrasonic Imaging Division, Renmin Hospital, Hubei University of Medicine, Shiyan, 442000, Hubei Province, China.
Tumour Biol. 2015 Sep;36(10):7997-8006. doi: 10.1007/s13277-015-3528-6. Epub 2015 May 14.
Glypican-3 (GPC3) is a membrane of heparan sulfate proteoglycan family involved in cell proliferation, adhesion, migration, invasion, and differentiation during the development of the majority of mesodermal tissues and organs. GPC3 is explored as a potential biomarker for hepatocellular carcinoma screening. However, as a tumor-associated antigen, its role in ovarian cancer remains elusive. In this report, the expression levels of GPC3 in the various ovarian cancer cells were determined with quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and GPC3 expression in ovarian cancer UCI 101 and A2780 cells was knocked down by siRNA transfection, and the effects of GPC3 knockdown on in vitro cell proliferation, migration, and invasion were respectively analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay and Transwell migration assay. Additionally, the effect of GPC3 knockdown on in vivo tumorigenesis were investigated in athymic nude mice. The results indicated that GPC3 knockdown significantly promoted cell proliferation and increased cell migration and invasion by upregulation of matrix metalloproteinase (MMP)-2 and MMP-9 expression and downregulation of tissue inhibitor of metalloproteinase-1 expression. Additionally, GPC3 knockdown also increased in vivo tumorigenicity of UCI 101 and A2780 cells and final tumor weights and volumes after subcutaneous cell injection in the nude mice. The results of immunohistochemical staining and Western blotting both demonstrated a lower expression of GPC3 antigen in the tumors of GPC3 knockdown groups than that of negative control groups. Moreover, transforming growth factor-β2 protein expression in the tumors of GPC3 knockdown groups was significantly increased, which at least contributed to tumor growth in the nude mice. Taken together, these findings suggest that GPC3 knockdown promotes the progression of human ovarian cancer cells by increasing their migration, invasion, and tumorigenicity, and suggest that GPC3 is a potential therapeutic target for ovarian cancer patients.
磷脂酰肌醇蛋白聚糖-3（GPC3）是硫酸乙酰肝素蛋白聚糖家族的一种膜蛋白，在大多数中胚层组织和器官发育过程中参与细胞增殖、黏附、迁移、侵袭及分化。GPC3被探索作为肝细胞癌筛查的潜在生物标志物。然而，作为一种肿瘤相关抗原，其在卵巢癌中的作用仍不清楚。在本报告中，采用定量逆转录-聚合酶链反应（qRT-PCR）测定不同卵巢癌细胞中GPC3的表达水平，通过小干扰RNA（siRNA）转染敲低卵巢癌UCI 101和A2780细胞中的GPC3表达，并分别采用3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）法和Transwell迁移试验分析敲低GPC3对体外细胞增殖、迁移及侵袭的影响。此外，在无胸腺裸鼠中研究敲低GPC3对体内肿瘤发生的影响。结果表明，敲低GPC3可通过上调基质金属蛋白酶（MMP）-2和MMP-9表达及下调金属蛋白酶组织抑制因子-1表达，显著促进细胞增殖并增加细胞迁移和侵袭。此外，敲低GPC3还增加了UCI 101和A2780细胞在裸鼠皮下注射细胞后的体内致瘤性以及最终肿瘤重量和体积。免疫组织化学染色和蛋白质印迹结果均显示，敲低GPC3组肿瘤中GPC3抗原的表达低于阴性对照组。此外，敲低GPC3组肿瘤中转化生长因子-β2蛋白表达显著增加，这至少促成了裸鼠体内肿瘤生长。综上所述，这些发现表明敲低GPC3通过增加人卵巢癌细胞的迁移、侵袭及致瘤性促进其进展，并提示GPC3是卵巢癌患者的潜在治疗靶点。
