Polo‐like kinase 1 inhibition suppresses hepatitis B virus X protein‐induced transformation in an in vitro model of liver cancer progression†

Abstract

Chronic hepatitis B virus (HBV) infection is linked to development of hepatocellular carcinoma (HCC). The HBV X protein (pX) is implicated in HCC pathogenesis acting as a weak oncogene or a cofactor in hepatocarcinogenesis. pX induces DNA re‐replication, DNA damage, and partial polyploidy in a poorly differentiated, immortalized hepatocyte cell line. In this study we employed sorted, pX‐induced polyploid cells to investigate their growth and oncogenic transformation potential over the course of 70 cell doublings. Immediately after live cell‐sorting, nearly 40% of pX‐induced polyploid cells undergo apoptosis, whereas the surviving cells exhibit proliferation sensitive to p53. After 40 cell generations the pX‐expressing polyploid cultures exhibit loss of p53 function and become growth factor‐ and anchorage‐independent, indicative of oncogenic transformation. The pX‐induced polyploid cultures in the course of 70 cell generations undergo progressively increasing DNA damage, propagate damaged DNA to daughter cells, and display increased expression of a cluster of proliferation genes shown to be elevated in human HCC, including HBV‐HCC. One of these genes is the mitotic kinase Polo‐like kinase 1 (Plk1). Oncogenic transformation is suppressed in the absence of pX expression, and significantly, by inhibition of Plk1. These results identify Plk1 as crucial in pX‐mediated oncogenic transformation. Conclusion: Partial polyploidy induced by pX is not immediately associated with oncogenic transformation. Continued DNA damage for 40 cell generations is reproducibly associated with loss of p53 function, enhanced expression of Plk1, and oncogenic transformation. Because Plk1 expression is also elevated in HBV‐HCC tumors, this in vitro cellular model simulates liver cancer progression and pathogenesis in chronic HBV patients. Inhibition of Plk1 activity suppresses pX‐mediated oncogenic transformation, identifying Plk1 as a promising therapeutic target for HBV‐mediated HCC. (HEPATOLOGY 2009.)