Reduction of GRP78 expression with siRNA activates unfolded protein response leading to apoptosis in HeLa cells

The 78-kDa glucose-regulated protein GRP78 is a central regulator of endoplasmic reticulum (ER) homeostasis, functioning in protein folding, ER calcium binding and modulation of transmembrane ER stress sensor activity. ER stress uncouples the interaction between GRP78 and ER stress sensors, leading to activation of the unfolded protein response (UPR), including upregulation of ER chaperone proteins. In the present study, we observed unexpected and remarkable induction of glucose-regulated protein 94 (GRP94) in HeLa cells following their transfection with 2′-O-methyl-modified siRNA specific to GRP78 mRNA. Additionally, we found that this siRNA also increased the expression of other UPR-induced genes, such as CHOP, ERdj4 and P5. Activation of UPR-dependent transcription and induction of apoptosis were also observed in cells transfected with GRP78 siRNA. Induction of apoptosis by GRP78 siRNA was also observed in PC-3 cells, which expressed high basal levels of GRP78 protein similar to that observed in HeLa cells. By contrast, five other human cell lines with lower basal expression of GRP78 protein did not undergo apoptosis when treated with GRP78 siRNA. Possible reasons for the strong activation of the UPR and apoptosis induced by GRP78 knockdown in HeLa cells, and the therapeutic utility of 2′-O-methyl-modified GRP78 siRNAs in anticancer treatment, are discussed.