Receptor mediated endocytosis and cytotoxicity of transferrin-mitomycin c conjugate in the HepG2 cell and primary cultured rat hepatocyte

Intracellular disposition and cytotoxicity of macromolecular conjugate of m itomycin C (MMC) with transferrin (TF) were examined in the human hepatoma cell line HepG2 cell and normal cultured rat hepatocyte. The conjugate (TF- MMC) was specifically bound to the HepG2 cell as well as TF, The number of the binding site and the association constant of TF-MMC in the HepG2 cell w ere 396000 +/- 31000 molecules/cell and 3.24 x 10(7) +/- 0.58 x 10(7) M-1, respectively No difference in the binding parameters of TF-MMC and TF can b e detected in the HepG2 tell. The association constant for the TF receptor was almost identical between HepG2 cell and hepatocyte, however, the number s of the binding site of TF-MMC and TF in the HepG2 cell were from 40-times to 50-times greater than those in the hepatocyte, Furthermore, TF-MMC was internalized into the HepG2 cell and the hepatocyte as well as TF The rates of internalization of TF-MMC and TF into the HepG2 cell were nearly identi cal to those into the hepatocyte. However, the levels of the internalizatio n into the HepG2 cell were remarkably higher than those into the hepatocyte because the number of receptors in the HepG2 cell was larger than that in the hepatocyte, and the rate of release from the HepG2 cell was slower than that from the hepatocyte, TF-MMC inhibited the growth of the HepG2 cells. The 50% growth inhibition (GI(50)) of TF-MMC against the HepG2 cell was 0.9 mug MMC/ml, which was a little higher than that of MMC (GI(50) = 0,5 mug/m l). These results indicated that the TF-MMC might be useful for delivery of MMC to the HepG2 cell.