Li Junping, Liu Yang, Yin Yihua
Department of Gynecology and Obstetrics, Huashan Hospital North, Fudan University, Shanghai, 200040, People's Republic of China.
Institute of Antibiotics,Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China.
Tumour Biol. 2016 Feb;37(2):1411-25. doi: 10.1007/s13277-015-4502-z. Epub 2015 Dec 1.
Ras homology GTPase activation protein 6 (Arhgap6), as a member of the rhoGAP family of proteins, performs vital functions on the regulation of actin polymerization at the plasma membrane during several cellular processes. The role of Arhgap6 in the progression and development of cancer remains nearly unknown. This study aimed at exploring the effects of Arhgap6 on cervical carcinoma. Human cervical cancer cells HeLa and SiHa were transduced with a lentivirus targeting Arhgap6 (Arhgap6+), while CaSki and C4-1 cells were transfected with miRNA. Cell proliferation was identified by Cell Counting Kit-8 (CCK-8). Cell cycle distribution and cell apoptosis were identified by flow cytometry. The capacity of cell migration, invasion, and adhesion were detected by Transwell assay. Further, quantitative real-time PCR (qRT-PCR) and western blot were used to analyze the expression levels of Arhgap6 and several tumor-related genes. Co-immunoprecipitation assay was performed to validate the interaction between Arhgap6 and Rac3 (Ras-related C3 botulinum toxin substrate 3). Results showed that Arhgap6 inhibited cell proliferation, migration, invasion, and adhesion of cervical carcinoma, induced cell apoptosis, and caused cell cycle arrest in the G0/G1 phase (n = 3, p < 0.05). Expression of the tumor suppressor genes and oncogenes were up- and down-regulated respectively by Arhgap6, and Rac3 was proved to be the target of Arhgap6. Besides, in in vivo assays, tumor size and weight were destructed in Arhgap6+ athymic nude mouse. This study indicated that Arhgap6 may play a role in the treatment of cervical cancer as a tumor supressor.
Ras同源GTP酶激活蛋白6（Arhgap6）作为rhoGAP蛋白家族的一员，在多个细胞过程中对质膜上肌动蛋白聚合的调节发挥着重要作用。Arhgap6在癌症进展和发展中的作用几乎仍不清楚。本研究旨在探讨Arhgap6对宫颈癌的影响。用靶向Arhgap6的慢病毒转导人宫颈癌细胞HeLa和SiHa（Arhgap6+），而用miRNA转染CaSki和C4-1细胞。通过细胞计数试剂盒-8（CCK-8）鉴定细胞增殖。通过流式细胞术鉴定细胞周期分布和细胞凋亡。通过Transwell实验检测细胞迁移、侵袭和黏附能力。此外，使用定量实时PCR（qRT-PCR）和蛋白质印迹分析Arhgap6和几个肿瘤相关基因的表达水平。进行免疫共沉淀实验以验证Arhgap6与Rac3（Ras相关的C3肉毒杆菌毒素底物3）之间的相互作用。结果显示，Arhgap6抑制宫颈癌细胞的增殖、迁移、侵袭和黏附，诱导细胞凋亡，并导致细胞周期停滞在G0/G1期（n = 3，p < 0.05）。Arhgap6分别上调和下调肿瘤抑制基因和癌基因的表达，并且证明Rac3是Arhgap6的靶点。此外，在体内实验中，Arhgap6+无胸腺裸鼠的肿瘤大小和重量减小。本研究表明，Arhgap6作为一种肿瘤抑制因子可能在宫颈癌治疗中发挥作用。
