Growth-promoting effect of di-2-ethylhexyl phthalate on human neuroblastoma cells / 复旦学报（医学版）

ABSTRACT

Objective To investigate the effect of di-2-ethylhexyl phthalate (DEHP) on the proliferation of SK-N-SH human neuroblastoma cells and its underlying mechanism. Methods Cells were cultured in estrogen-free improved Dulbecco's Modified Eagle's Medium and then divided into 5 groups: no treatment (control group); treated with 17β-estradiol (E_2 group); treated with DEHP (DEHP group); treated with both E_2 and phosphatidylinositol-3-kinases (PI3K) inhibitor LY294002 (E_2 + LY294002 group); treated with both DEHP and LY294002 (DEHP + LY294002 group). The absorbance value (AV) was measured on day 0, 2, and 5. DNA proliferation index (PI) and apoptotic index (AI) were determined by flow cytometry on day 5. Caspase-3 protein, protein-serine-threonine kinase (Akt) and phosphor-Akt (Ser473) protein expression were analyzed by Western blot on day 5. Results The AV of All groups increased on day 2, and 5. The AV of E_2 and DEHP groups were higher than that of the control group (P<0. 001), but the AV of E_2 + LY294002 and DEHP + LY294002 groups were lower than those of E_2 and DEHP groups (P<0.01) on day 2 and 5. On day 5, PI of E_2 and DEHP groups were also higher than that of control (P<0.01). However, PI of E_2 + LY294002 and DEHP + LY294002 groups were lower than those of E_2 and DEHP group (P<0.01) on day 5. There was no significant difference in AI and caspase-3 protein expression among the groups. At the same time, phosphor-Akt (Ser473) protein expression of E_2 and DEHP groups increased obviously, compared with the control group. Compared with E_2 and DEHP groups, E_2 + LY294002 and DEHP + LY294002 groups decreased significantly. However, Akt protein expression was equal among those groups. Conclusions DEHP can promote the growth of SK-N-SH cells to a level similar to that of E_2, with activation of the PI3K/Akt signaling pathway.