Differential expression of the Kv3.4 channel in in vitro and in vivo models of cervical cancer: Potential clinical implications

Introduction: Cervical cancer remains a major global health concern, and current diagnostic methods have significant limitations, highlighting the need for improved molecular biomarkers. Its progression is primarily linked to HPV infection, estrogen exposure, and ion channel deregulation. Ion channels such as Kv3.4 have gained relevance due to their involvement in carcinogenesis. This study examines the gene and protein expression of the Kv3.4 (KCNC4) channel in cervical cancer models and evaluates the effects of its pharmacological blockade as a potential diagnostic and therapeutic strategy. Experimental procedure: KCNC4 gene expression was study in HeLa and SiHa cells by qRT-PCR, while Kv3.4 protein was evaluated by immunocytochemistry in HeLa, SiHa, normal keratinocytes (KN) and keratinocytes with HPV16 E6/E7 oncogenes (KT E6E7). The effect of Kv3.4 functional inhibition was assessed by metabolic activity assays following treatment with the Kv channel blocker BDS-I. In vivo model from four mouse groups: transgenic K14E7 mice treated with 17beta-estradiol (K14E7+E2), untreated K14E7 mice, non-transgenic FvB mice treated with 17beta-estradiol (FvB+E2), and untreated FvB controls, were examined by immunohistochemistry to determine the impact of HPV oncogenes and estrogen exposure on Kv3.4 regulation in cervical epithelium. Results: Gene expression analysis showed a significantly higher KCNC4 expression in HeLa cells compared with SiHa cells, consistent with their differing HPV copy numbers. Immunocytochemistry confirmed Kv3.4 protein expression in HeLa, SiHa, and KT E6E7 cells, but not in KN cells. BDS-I treatment significantly reduced metabolic activity only in KT E6E7 cells, with no effect in KN cells. In mouse cervical tissues, Kv3.4 expression was strongest in FvB+E2 mice, followed by K14E7 and untreated FvB at 4 months. At 7 months, expression remained highest in FvB+E2, then K14E7+E2, K14E7, and FvB, indicating that estrogen exposure and HPV oncogenes enhance Kv3.4 expression in vivo. Conclusion. Our findings show that the Kv3.4 channel is linked to HPV-related cervical carcinogenesis. Kv3.4 protein was detected in HeLa, SiHa, KT E6E7, and in cervical tissues from FvB+E2, K14E7+E2, and K14E7 mice, indicating that estrogen and HPV oncogenes upregulate the channel in vivo. Notably, Kv3.4 blockade selectively reduced proliferation in KT E6E7 cells, suggesting a greater dependence of HPV-transfected keratinocytes on this channel. Overall, these results support Kv3.4 as a potential biomarker and therapeutic target in early HPV-mediated cervical cancer. Partially supported by Conacyt grant A1-S-9783 to JdlG.