IL-37 attenuated HPV induced inflammation and growth of oral epithelial cells via regulating autophagy

Abstract

This study investigated the impact of Human Papillomavirus (HPV) on inflammation and growth in oral epithelial cells, with a focus on the role of Interleukin-37 (IL37). Oral epithelial cells, including HOEC and HSC-3 cells, were employed in the research. The results revealed that HPV significantly induced inflammation in both types of oral epithelial cells, concurrently promoting cell growth and inhibiting apoptosis. IL37, a cytokine, was found to mitigate HPV-induced inflammation in oral epithelial cells. Moreover, IL37 counteracted HPV's effects on apoptosis and cell viability in oral epithelial cells. The study also identified a reduction in autophagy in HPV-infected oral epithelial cells, a phenomenon alleviated by IL37. Furthermore, chemical inhibition of autophagy was observed to attenuate HPV-induced inflammation and growth in oral epithelial cells. These findings contribute valuable insights into the pathogenesis of inflammation in oral epithelial cells associated with HPV and oral cancers, offering potential avenues for novel therapeutic strategies.

Keywords: Growth; HPV; IL-37; Inflammation; Oral epithelial cells.

Fig. 1

HPV E6 significantly induces inflammation in oral cells. (A) HPV E6 significantly increased expression of IL6 in HOEC cells in dose dependent manner; (B) HPV E6 significantly increased expression of IFNβ in HOEC cells in dose dependent manner; (C) HPV E6 significantly increased expression of IL10 in HOEC cells in dose dependent manner; (D) HPV E6 significantly increased expression of IL6 in HSC-3 cells in dose dependent manner; (E) HPV E6 significantly increased expression of IFNβ in HSC-3 cells in dose dependent manner; (F) HPV E6 significantly increased expression of IL10 in HSC-3 cells in dose dependent manner.

Fig. 2

HPV E6 significantly promoted growth and decreased apoptosis of oral epithelial cells. (A) HPV E6 significantly increased cell viability of HOEC cells in a dose dependent manner; (B) HPV E6 significantly increased cell viability of HSC-3 cells in a dose dependent manner; (C) HPV E6 significantly decreased expression level of caspase 3 on HOEC cells in a dose dependent manner; (D) HPV E6 significantly decreased expression level of caspase 3 on HSC-3 cells in a dose dependent manner; (E) HPV E6 significantly decreased expression level of caspase 7 on HOEC cells in a dose dependent manner; (F) HPV E6 significantly decreased expression level of caspase 7 on HOEC cells in a dose dependent manner; (G) HPV E6 significantly decreased expression level of caspase 9 on HOEC cells in a dose dependent manner; (H) HPV E6 significantly decreased expression level of caspase 9 on HOEC cells in a dose dependent manner.

Fig. 3

IL37 attenuated HPV induced inflammation on oral epithelial cells. (A) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of IL17 protein level on HOEC cells; (B) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of IL17 protein level on HSC-3 cells. (C) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of IL6 protein level on HOEC cells; (D) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of IL6 protein level on HSC-3 cells; (E) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of TNFɑ protein level on HOEC cells (F) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of TNFɑ protein level on HSC-3 cells.

Fig. 4

IL37 attenuated HPV promoted growth and decreased apoptosis on oral tumor cells. (A) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced decrease of casepase 3/7 activity on HOEC cells; (B) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced decrease of casepase 3/7 activity on HSC-3 cells; (C) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of apoptosis on HOEC cells. (D) IL37 (5 and 20 ng/mL) significantly attenuated HPV E6 induced increase of apoptosis on HSC-3 cells; (E) IL37 (20 ng/mL) significantly attenuated HPV E6 induced decrease of apoptosis on HOEC and HSC-3 mixed cells evaluated by FACS.

Fig. 5

IL37 attenuated HPV diminished autophay on oral epithelial cells. (A) HPV E6 significantly decreased expression of ATG8 on HOEC cells; (B) HPV E6 significantly decreased expression of P62 on HOEC cells; (C) HPV E6 significantly decreased expression of ATG8 on HSC-6 cells; (D) HPV E6 significantly decreased expression of P62 on HSC-3 cells; (E) HPV E6 significantly decreased protein level of ATG8 on HOEC cells; (F) HPV E6 significantly decreased protein level of ATG8 on HSC-3 cells.

Fig. 6

Increase of inflammation and decrease of apoptosis induced by HPV was through authopahgy. (A) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced increase of expression level IL17A on HOEC cells; (B) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced increase of expression level IL17A on HSC-3 cells; (C) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced increase of expression level IL6 on HOEC cells; (D) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced increase of expression level IL6 on HSC-3 cells; (E) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced increase of expression level TNFɑ on HOEC cells; (F) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced increase of expression level TNFɑ on HSC-3 cells; (G) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced decrease of caspase3/7 activity on HOEC cells; (H) Autophagy inhibitor 3-Methyladenine (3-MA) significantly inhibited HPV E6 induced decrease of caspase3/7 activity on HSC-3 cells.

figs1

figs2

figs3

figs4

figs5