The Effect Of Periplocin From Cortex Periplocae And TRAIL On Gastric Cancer Cells And The Mechanism

Objective:To investigate the effect of periplocin and combination with TRAIL on proliferation of gastric cancer cells and to explore the mechanismMethods:1 The proliferation of gastric cancer cells treated with periplocin and combination with TRAIL was detected by MTS.2 Flow cytometry, TUNEL assay and Hochest assay detects periplocin and combination with TRAIL inducing apoptosis of gastric cancer.3 The expression levels of apoptosis related proteins were detected by western blotting.4 Acridine orange staining and immunofluorescence detects periplocin inducing apoptosis by lysosomal pathway.5 Gene chip detects the expression levels of genes on gastric cancer cells which treated with periplocin and proved by qPCR.6 The expression levels of EGR1 and p-ERK1/2 on gastric cancer cells which treated with periplocin and combination with TRAIL were detected by Western blotting.7 The proliferation of first treated with periplocin after knockdown EGR1 on gastric cancer cells were detected by MTS.8 In BALB/c nude mice detect the effect on growth viability of gastric cancer cells treated with periplocin and combination with TRAIL.9 TUNEL assay detects periplocin and combination with TRAIL inducing apoptosis of gastric cancer cells in vivo.10 Immunohistochemistry detects the expression levels of p-ERK1/2, EGR1, Ki-67, DR4,DR5,caspase-3,Bid,Mcl-1 on gastric cancer cells treated with periplocin and combination with TRAIL.Result:1 MTS assay demonstrated that periplocin and combination with TRAIL inhibited the proliferation of gastric cancer cells SGC-7901, MGC-803 and BGC-823, and in a time and dose dependent(P<0.05).2 Flow cytometry, TUNEL assay and Hochest assay demonstrated periplocin and combination with TRAIL induced apoptosis of gastric cancer(P<0.05).3 Western blotting demonstrated periplocin and combination with TRAIL increased the expression levels of cleaved caspase-3 and decreased the expression of Mcl-1 and pro-Bid(P<0.05).4 Acridine orange staining and immunofluorescence demonstrated periplocin destroyed the integrity of the lysosomal membrane and Cathepsin B, CathepsinD released into cytoplasm.5 Gene chip demonstrated periplocin from cortex Periplocae increased the expression of EGR1 on gastric cancer cells.6 Western blotting demonstrated periplocin from cortex Periplocae and TRAIL and increased the expression of p-ERK1/2, EGR1 on gastric cancer cells(P<0.05).7 MTS assay demonstrated that the proliferation of first treated with periplocin after knockdown EGR1 on gastric cancer cells was not changed(P>0.05).8 The in vivo experiment demonstrated periplocin and combination with TRAIL inhibited the growth of xenografts(P<0.05).9 TUNEL assay demonstrated periplocin and combination with TRAIL induced apoptosis of gastric cancer cells in vivo(P<0.05).10 Immunohistochemistry demonstrated the expression levels of EGR1,p-ERK1/2,DR4,DR5 on gastric cancer cells treated with periplocin and combination with TRAIL in vivo were increased,the levels of Mcl-1, pro-Bid, pro-caspase-3 were decreased.Conclusions:1 Periplocin and combination with TRAIL inhibit the proliferation of gastric cancer cells by induction of apoptosis.2 Periplocin may be induce apoptosis by lysosomal pathway.3 Periplocin and combination with TRAIL inhibit the proliferation of gastric cancer cells by increasing the expression levels of EGR1 and ERK1/2.