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

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.

Downregulation of KCTD12 contributes to melanoma stemness by modulating CD271

“…In addition, KCTD12 was widely reported to be downregulated in gastrointestinal stromal tumors (GISTs) and served as a powerful prognostic marker for GISTs 56 . However, few studies have focused on the role of KCTD12 in melanoma metastasis 6 . In the present study, we analyzed the correlation between KCTD12 expression levels and the clinical prognosis of SKCM patients using the GEPIA database.…”

“…The underlying mechanism through which KCTD12 downregulation promoted metastasis was preliminarily investigated. KCTD12 has been reported to suppress uveal melanoma cell proliferation by prolonging G2/M to G1 phase progression 6 . Earlier reports stated that KCTD12 inhibited CRC stemness and metastasis through suppressing the ERK pathway 5 .…”

“…A previous study reported that colorectal cancer stemness was regulated by KCTD12 through the ERK pathway 5 . Earlier reports declared that KCTD12 promoted the proliferation of human uveal melanoma OCM-1 cells 6 . In addition, it was reported that KCTD12 acted as a tumor suppressor in esophageal squamous cell carcinoma (ESCC) through the WNT/NOTCH pathway and chromatin remodeling 7 .…”

Melanoma proteomics unravels major differences related to mutational status

“…Another protein differentially expressed is the coagulation factor XIII (F13A1) which it was previously associated with chemotherapy response in melanoma tumors [22]. Potassium channel tetradimerization domain containing 12 (KCTD12) inhibits proliferation in uveal melanoma cells [23]. SLC9A3R1 is involved in suppressing breast cancer cells proliferation [24].…”

Proteomic analysis of RAW macrophages treated with cGAMP or c-di-GMP reveals differentially activated cellular pathways

“…50 Luo et al demonstrated that the over expression of KCTD12 in human uveal melanoma OCM-1 cells inhibited proliferation. 51 In another report, Li et al showed that colorectal cancer cells stemness was regulated by KCTD12 via the ERK pathway. 50 Retinoic acid early-inducible protein 1-gamma, RAE-1c (4.04 fold) was also upregulated in the c-di-GMP treatment group.…”