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We have shown previously that loss of protein kinase D (PKD) activity in individual cell types can sensitize them to the growth inhibitory effects of the active phospholipid-dependent protein kinase 1 (PLD1). In this report, we address the functional significance of the resultant PLD1-PKD-mediated growth arrest. Unlike wild-type human lung cancer cell lines, loss of PLD1 activity in H1299 and H460 cells resulted in cell-cycle arrest in G(1) phase. This was accompanied by up-regulation of P21(cip1), with a lesser effect on P16(INK4a), and a significant increase in the expression of the CKI p16(INK4a), which is generally low in lung cancers. However, the suppression of PLD1 activity in H1299 cells did not alter E2F levels in the cell. Expression of a dominant negative PKD construct blocked the cell-cycle arrest and abrogated the G(1) arrest associated with loss of PLD1 activity. We also show that the PKD-mediated cell-cycle arrest is mediated through loss of the active PKD1, since cell-cycle analysis revealed arrest in G(1) phase in H1299 cells infected with adenovirus expressing dominant negative PKD1. These results were corroborated by cell cycle analysis following overexpression of constitutively active PKD1 in wild-type and PLD1-knockdown H1299 cell lines. Our data identify PLD1 and PKD1 as functionally independent effectors of the G(1) arrest in human lung cancer.Where would you like to sign in?

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