High-risk human papillomaviruses (HPVs) are the primary causes of cervical cancer. This study investigates the mechanism by which HPV16 E6 promotes cervical cancer cell proliferation by activating the pentose phosphate pathway (PPP). The authors found that HPV16 E6 increases glucose-6-phosphate dehydrogenase (G6PD) enzyme activity by inhibiting its lactylation, leading to increased NADPH and GSH levels and reduced ROS levels. In vitro and in vivo experiments demonstrated that inhibiting G6PD activity or mimicking lactylation through mutations in G6PD K45A significantly reduced cell proliferation and tumor growth. These findings suggest that HPV16 E6 activates the PPP to promote cervical cancer cell proliferation, providing a novel therapeutic target for HPV-associated cancers.High-risk human papillomaviruses (HPVs) are the primary causes of cervical cancer. This study investigates the mechanism by which HPV16 E6 promotes cervical cancer cell proliferation by activating the pentose phosphate pathway (PPP). The authors found that HPV16 E6 increases glucose-6-phosphate dehydrogenase (G6PD) enzyme activity by inhibiting its lactylation, leading to increased NADPH and GSH levels and reduced ROS levels. In vitro and in vivo experiments demonstrated that inhibiting G6PD activity or mimicking lactylation through mutations in G6PD K45A significantly reduced cell proliferation and tumor growth. These findings suggest that HPV16 E6 activates the PPP to promote cervical cancer cell proliferation, providing a novel therapeutic target for HPV-associated cancers.