Abstract No. 468 IL-17 treatment in nutritionally deprived condition decreases cell migration in rat hepatoma McA-RH7777

Abstract

To simulate postembolization environment in vitro, and evaluate cellular morphological and biological changes from IL-17 treatment using a hepatocellular carcinoma cell line. The hepatoma cell line McA-RH7777 was used in this study. The migration potential of cells was observed using wound healing assay, and proliferation potential was investigated using colony formation assay. In order to simulate nutrient-depleted environment after embolization and quantitatively measure postembolization changes in vitro, rat hepatoma McA-RH7777 cell line was exposed to low glucose culture media. Wound healing assay and colony formation assay were conducted under low and normal glucose condition with and without IL-17 treatment. For wound healing assay, cells were plated with >90% confluency before initiation of scratch. Serial pictures were taken and wound area was measured in pixels using ImageJ. For colony formation assay, 500 cells per well in 6-well plate were plated as single cell suspension and grown to form colony. After 12days plates were fixed, stained and only colonies with >50 cells were considered as colonies to be counted. Statistical analysis was performed using Graphpad prism 7.03 software. Migration assay indicated a decrease in migration potential in IL-17 treated and exposed in low nutritional environment. Result showed statistically significant difference in 24 hours (Low glucose DMEM 19.47 vs IL-17 treated Low glucose DMEM 23.69, p=0.003) and 48 hours (7.13 vs 15.34, p=0.001) from initial scratch. Colony formation assay showed higher proliferation potential in low glucose media compared to regular glucose media (183.3 vs 142.7, p=0.004). However, no significant difference found with and without IL-17 treatment in both nutritional environments. The low nutritional environment in the tumor cell line, simulating postembolization changes, compared to normal nutritional environment increases proliferative potential. However, in nutritionally deprived condition, IL-17 decreases migration. Decreased migration and increased proliferation potential of residual tumors of HCC may be responsible for local tumor recurrence after transarterial embolization.