Lentiviral-mediated overexpression of KCTD12 inhibits the proliferation of human uveal melanoma OCM-1 cells

Luo,Lifu; Cui,Jizhe; Feng,Zhitong; Li,Yana; Wang,Mengdi; Cai,Yong; Wu,Yazhen; Jin,Jingji

doi:10.3892/or.2016.5325

Lentiviral-mediated overexpression of KCTD12 inhibits the proliferation of human uveal melanoma OCM-1 cells

Authors:
- Lifu Luo
- Jizhe Cui
- Zhitong Feng
- Yana Li
- Mengdi Wang
- Yong Cai
- Yazhen Wu
- Jingji Jin
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Affiliations: Department of Ophthalmology, The Second Hospital of Jilin University, Changchun, Jilin 130041, P.R. China, School of Life Sciences, Jilin University, Changchun, Jilin 130012, P.R. China
Published online on: December 16, 2016 https://doi.org/10.3892/or.2016.5325
Pages: 871-878

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Abstract

Human potassium channel tetramerization domain containing 12 (KCTD12, also known as Pfetin) is a member of the KCTD family which consists of 26 members. It has been reported that KCTD12 regulates agonist potency and kinetics of GABAB receptor signaling. Proteomic analysis indicates that KCTD12 may be a potential biomarker for the diagnosis and prognosis of gastrointestinal stromal tumors. However, little has been reported concerning the role of KCTD12 in the other tumor types. In the present study, we designed and subcloned N-terminally Flag-tagged human KCTD12 into the pLVX‑Puro vector. We then generated a human uveal melanoma cell line (OCM-1) stably expressing KCTD12. Using this stable cell line, we performed a series of experiments including colony formation, invasion, migration and wound-healing assays, flow cytometry and western blotting. Based on the experimental results, we first demonstrated that KCTD12 effectively suppressed the proliferation of OCM-1 cells and limited the spread of OCM-1 cells. In the flow cytometric analysis, prolongation of the progression of G2/M to G1 phase in the KCTD12-overexpressing OCM-1 cells was observed. In addition, inhibition of KCTD12-overexpressing OCM-1 cell xenograft growth in nude mice was observed. Taken together, KCTD12 may serve as a novel therapeutic target for patients with uveal melanoma.

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