Silencing of voltage-gated potassium channel K_V9.3 inhibits proliferation in human colon and lung carcinoma cells

Voltage-gated potassium (K_v) channels are known to be involved in cancer development and cancer cell proliferation. K_V9.3, an electronically silent subunit, forms heterotetramers with K_V2.1 in excitable cells and modulates its electrophysiological properties. However, the role of K_V9.3 alone in non-excitable cancer cells has not been studied. Here, we evaluated the effect of silencing K_V9.3 on cancer cell proliferation in HCT15 colon carcinoma cells and A549 lung adenocarcinoma cells. We confirmed the expression of K_V9.3 mRNA in HCT15 and A549 cells and showed that silencing K_V9.3 using small interfering RNA caused G0/G1 cell cycle arrest and alterations in cell cycle regulatory proteins in both HCT15 and A549 cells without affecting apoptosis. Also, stable knockdown of K_V9.3 expression using short-hairpin RNA inhibited tumor growth in SCID mouse xenograft model. Using a bioinformatics approach, we identified Sp1 binding sites in the promoter region of the gene encoding K_V9.3. We further found that Sp1 bound to this region and showed that the Sp1 inhibitor, mithramycin A, induced a concentration-dependent decrease in K_V9.3 expression. Taken together, these data suggest that knockdown of K_V9.3 inhibits proliferation in colon carcinoma and lung adenocarcinoma cell lines and may be regulated by Sp1.