PC-3 cells express functional GJICs.

<p>(A) Expression of GJIC components, connexin43 and pannexin1, was confirmed by Western blot on whole-cell lysates of non-transduced, LV/TMPK-transduced, and LV/eGFP-transduced PC-3 cells. GAPDH expression was evaluated as an equal loading control. (B) Expression of TMPK was confirmed by Western blot on whole-cell lysates of non-transduced, LV/TMPK-transduced, and LV/TMPK-F105Y-transduced PC-3 cells. GAPDH expression was evaluated as an equal loading control. (C) Expression pattern of connexin43 (left panel) and pannexin1 (right panel) GJIC components (green fluorescence) was assessed by confocal immunofluorescent microscopy. Cytoskeleton (F-actin) was visualized with rhodamine phalloidin (red fluorescence) staining. (D) Dye transfer of calcein into adjacent PKH26-labelled cells was measured by flow cytometry in direct (normal) or transwell co-cultures of PC-3 cells as percentage of cells double-positive for calcein and PKH26 fluorescence (n=3). (E) Dye transfer of calcein into adjacent PKH26-labelled cells was significantly inhibited by carbenoxolone (CBX) (n=3). Statistical significance is indicated (p<0.0001).</p

Bystander cell killing mediated by TMPK-F105Y/AZT therapy drives a significant tumor mass reduction in a prostate cancer xenograft model.

<p>(A) Magnitude of bystander cell killing was evaluated following the delivery of the TMPK-F105Y suicide gene by direct intratumoral injection of LV/TMPK into established tumors in NOD/SCID mice (n=6). Reduction in tumor mass was assessed at the end of 6-day AZT treatment regimen (at 50mg/kg/day) by extraction of tumors and measurement of wet tumor weight. Statistical significance is indicated by an asterisk (* p<0.05). (B) Weight of individual extracted tumors is shown for each animal in the AZT-treated and vehicle-untreated groups.</p