Effect of interleukin-17A on stemness of hepatoma cell lines

Objective To investigate the effect of interleukin-17A (IL-17A) on stemness of human hepatoma cell lines Hep 3B, MHCC97 H, and MHCC97 L and the association between IL-17 A and the progression of liver cancer. Methods Human hepatoma cell lines Hep 3B, MHCC97 H, and MHCC97 L were selected, and in vitro 3D sphere formation assay was used to analyze the effect of IL-17 A on sphere formation ability. The control group with common culture solution and the experimental group with 50 ng/ml IL-17 A were established. Real-time cellular analysis was used to determine the effect of IL-17 A on the proliferation and migration of hepatoma cells with enhanced sphere formation ability; quantitative real-time PCR was used to measure the changes in the mRNA expression of IL-17 A receptors IL-17 RA and IL-17 RC and stemness-related genes SOX2, NANOG, OCT4, and BMI1 in hepatoma cells with enhanced sphere formation ability; Western blot was used to measure the expression of epithelial-mesenchymal transition-related proteins E-cadherin, N-cadherin, and vimentin. The t-test was used for comparison of continuous doota betwwen groups. Results With the presence of 50 ng/ml IL-17 A and 500 inoculated cells, Hep 3B cells had a significant increase in the number of spheres formed (113. 0 ± 10. 3 vs 180. 0 ± 7. 2, t =5. 533, P < 0. 001) , while MHCC97 H and MHCC97 L cells showed no significant changes (t = 1. 087 and 0. 279, P = 0. 325 and 0. 785) .The analysis showed that IL-17 A promoted the proliferation and migration of Hep 3B cells with an increased number of spheres formed. After the addition of 50 ng/ml IL-17 A, there was an increase in the mRNA expression of IL-17 A receptors IL-17 RA and IL-17 RC over the time of treatment; Hep 3B cells showed significant increases in the mRNA expression of stemness-related genes SOX2 (t = 4. 749, P =0. 042) , NANOG (t = 19. 600, P = 0. 003) , OCT4 (t = 37. 310, P < 0. 001) , and BMI1 (t = 16. 810, P = 0. 004) . Western blot showed no significant change in the expression of the epithelium-derived marker E-Cadherin; there was an increase in the expression of the interstitial marker N-cadherin after the treatment with 50 ng/ml IL-17 A for 48 hours (t = 17. 620, P = 0. 036) ; there was no significant change in the expression of vimentin. Conclusion To a certain degree, IL-17 A enhances the stemness of Hep 3B hepatoma cells.