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To address this possibility, asbt was expressed in McArdle RH-7777 cells and then tested for response to glycine- and taurine-conjugated bile acids.
24, which were elevated compared to that in McArdle RH-7777 cells (Figure 10B).
This leaves the possibility that conjugated bile acids may activate the apoptotic cascade through Fas receptor in this cell line, as glycine-conjugated bile acids do in hepatocytes [ 25 ] . However, the activation of caspases does not require a specific kind of bile acid transporter since TCA is not toxic to McArdle RH-7777 cells irrespective of whether these cells internalize bile acids via ntcp or asbt.
The human bile acid binder (HBAB) cDNA [ 37 ] was cloned into a derivative of pCep4 (Invitrogen Corp., Carlsbad, CA), that had been previously modified by deleting the EBNA 1 gene [ 38 ] . The lipofectamine reagent (Canadian Life Technologies) was used to transfect the expression plasmid into CHO, McArdle RH7777, and McNtcp.
Treatment of McArdle RH-7777 and CHO cells with taurine- or glycine-conjugated bile acids had little effect on the release of LDH (Figure 4Cand 4D), indicating that bile acid uptake activity is necessary to manifest their cytotoxic potential in both cell lines.