Fig. 4: Ectopic overexpression of HBO1 exerts pro-cancerous activity in primary CRPC cells.

The primary pPC-1 cells were transduced with a lentiviral HBO1-expressing construct, and two stable selections, oeHBO1-Slc-1 and oeHBO1-Slc-2, were formed after selection and overexpression verification. Control pPC-1 cells were stably transduced with the empty vector (“Vec”); Expression of HBO1 mRNA and listed proteins was tested (A–C). Cells were cultivated for the designated hours, and cell proliferation and in vitro cell migration were tested by nuclear EdU incorporation (D) and “Transwell” (E) assays, respectively. The primary human prostate cancer cells derived from three other CRPC patients, pPC-2/pPC-3/pPC-4 (F–H), or the primary human prostate epithelial cells (“pEpi1” and “pEpi2”, derived from two patients) (I, J), were stably transduced with the lentiviral HBO1-expressing construct (“oeHBO1”) or the empty vector (“Vec”), HBO1 mRNA expression was measured (F, I). After culturing for indicated time periods, cell proliferation (G), in vitro migration (H) and cell viability (J) were measured similarly. Data were expressed as the mean ± standard deviation (SD, n = 5). *P < 0.05 versus “Vec” group. “n. s.” stands for non-statistical difference (P > 0.05). Scale bar = 100 μm.