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Hepatitis B virus X protein, and polyploidy in the development of HCC
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Hepatitis B Virus (HBV) X protein (pX) is implicated§by an unknown mechanism in§hepatocellular carcinoma (HCC) development,§characterized by increased rate of§chromosomal aberrations. Although most patients clear§the infection and develop§immunity, chronic infection in 5 to 10% infected§patients lead to HCC development in§the 4th to 5th decade of life. I demonstrate that§tetracycline-regulated pX expression§induces multipolar spindles and polyploidy ( 4N DNA).§To understand the mechanism§of pX-mediated polyploidy, I investigated whether pX§promotes DNA re-replication.§Dual-parameter flow cytometry demonstrates§pX-dependent BrdU incorporation in cells§with 4N DNA. pX also induces expression of§replication initiation factors Cdc6 and§Cdtl, while suppressing geminin expression, a§negative regulator of re-replication. G2-§phase synchronized cells exhibit pX-dependent: i)§nuclear Cdc6 and Mcm5 colocalization;§ii) absence of nuclear geminin; iii) increased BrdU§incorporation; iv) ATR§activation; v) RAD17 and H2AX phosphorylation; and§vi) co-localization of -H2AX§with the DNA elongation factor PCNA.
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