Involvement of endoplasmic reticulum in paclitaxel-induced apoptosis

The participation of the mitochondrial pathway in paclitaxel-incluced apoptosis has been well documented. After addition of paclitaxel to U937 cells, however, we observed an early expression of five endoplasmic reticulum (ER) stress response genes that preceded the release of cytochrome cfrom the mitochondria and the cleavage of the caspases. Involvement of the ER was supported by the following evidence. Paclitaxel treatment not only activated calpain and caspase-4, but also induced a gradual increase in the cytosolic Ca2+ concentration at 3-6 h. Paclitaxel-induced apoptosis can be inhibited by the calpain inhibitor calpeptin and IP3 receptor inhibitors. Either buffering of the cytosolic Ca2+ or inhibition of rnitochondrial calcium uptake reduced BiP expression. These inhibitors also reduced mitochondrial apoptotic signals, such as mitochondrion membrane potential disruption, cytochrome c release and eventually reduced the death of U937 cells. Paclitaxel-incluced Bax/Bak translocation to the ER and Bax climerization on the ER membrane occurred within 3 h, which led to a Ca2+ efflux into cytosol. Moreover, we found that cytochrome c translocated to the ER after releasing from mitochondria and then interacted with the IP3 receptor at 12-15 h. This phenomenon has been known to amplify apoptotic signaling. Taken together, ER would seem to contribute to paclitaxelinduced apoptosis via both the early release of Ca2+ and the late amplification of mitochondria-mediated apoptotic signals.