Loss of AMPK B1 enhances ovarian cancer cell growth Seliciclib CDK2 and anchorage Inhibitors,Modulators,Libraries independent growth ability Because AMPK B1 was obviously reduced in advanced stage ovarian cancer, we investigated the effect of AM PK B1 on ovarian cancer cell growth and anchorage independent growth. Stable clones overexpressing AMPK B1 in two ovarian cancer cell lines with relatively lower AMPK B1 level or depleted of AM PK B1 by shRNAi mediated gene silencing in another two ovarian cancer cell lines with relatively higher AMPK B1 expression were generated. The XTT cell proliferation assay demonstrated that enhanced expression of AMPK B1 significantly inhibited ovarian cancer cell growth by 45 to 50% in A2780cp and SKOV3 stable clones compared with the parental lines and vector controls.

Further more, transient upregulation of AMPK B1 elevated pAM PK and mitigated cell proliferation in ovarian cancer cells in a dose dependent manner. Additionally, we demonstrated that enforced expression of AMPK B1 exhibited Inhibitors,Modulators,Libraries 60 to 70% less foci in A2780cp and SKOV3 stable clones by the focus formation assay, and we demonstrated that the AMPK B1 overexpressed clones of A2780cp showed a 70% to 75% reduction in the number and size of colonies compared with the vector controls Inhibitors,Modulators,Libraries by the focus formation assay. Conversely, by depleting en dogenous AMPK B1 in OV2008 and OVCA 433 cells, which highly express AMPK B1, using the sh B1 shRNA, we demonstrated that cell prolif eration increased 20 25% in all stable clones that overex pressed the sh B1 shRNA.

Similarly, the stable AMPK B1 knockdown clones exhibited a 2 3 fold increase in cell growth based on the focus formation assay and a 4 5 fold increase in colony for mation using the anchorage independent growth ability assay. Given that Inhibitors,Modulators,Libraries overexpression of AMPK B1 could inhibit ovarian cancer cell growth, we investigated how AMPK B1 affected the cell cycle kinetics of ovarian cancer cells. We then demonstrated that overexpression of AMPK B1 induced G1 phase arrest in A2780cp and SKOV3 stables clones compared to the controls by a cell cycle analysis using flow cytometry. On the other hand, stable knock down of endogenous AMPK Inhibitors,Modulators,Libraries B1 enhanced the G1 phase in OV2008 and OVCA433 cells. In sum, these findings suggest that AMPK B1 plays a sup pressive role in the cell growth and anchorage independent growth capacity of ovarian cancer cells by inducing G1 phase arrest.

Loss of AMPK B1 promotes ovarian cancer cell migration and invasion We also studied the functional role of AMPK B1 in ovar ian cancer cell migration and invasion. Using transwell migration and invasion assays, enhanced AMPK B1 ex pression product information was found to significantly attenuate the cell mi gration and invasive capacities of SKOV3 stable clones. In contrast, stable depletion of endogenous AMPK B1 in AMPK B1 expressing OVCA433 cells using the sh B1 shRNA enhanced cell migration and invasion.