SiRNA inhibit glucose regulative proteins 78 and glucose regulative proteins 94 gene and its effect on cell proliferation and apoptosis of human gastric cancer cell line in vitro

Abstract

Objective To study the effect of psiSTRIKETM/glucose regulative proteins 78 (GRP78) and psiSTRIKETM/glucose regulative proteins 94 (GRP94) on the expression of GRP78 and GRP94 in human gastric cancer cell line SGC7901 and its effect on cell proliferation and apoptosis in vitro.Methods PsiSTRIKETM/GRP78 and psiSTRIKETM/GRP94 were transfected to gastric cancer cell by LipofectaminTM 2000.The expression of GRP78 and GRP94 mRNA were analyzed by reverse transcriptasepolymerase chain reaction (RT-PCR) and the protein of GRP78 and GRP94 were detected by immunofluorescence 72h after transfection.Cells proliferation was detected by methyl thiazolium tetrazolium (MTT) and the apoptosis of gastric cancer cell was detected by Flow Cytometry.Results The result of RT-PCR showed that GRP78 (0.49) and GRP94 (0.40)of experimental group were declined obviously compare with control groups.and immunofluorescence showed that the expression of GRP78 and GRP94 of gastric cancer cell were inhibited obviously 72h after transfection.The results of MTT showed that compared with control groups,the proliferation of gastric cancer cell in experiment group was inhibited obviously 72 h after transfection (P ＜ 0.05).The ratio of apoptosis was significantly increased in experiment groups (21.98％) compared with control groups (P ＜ 0.05).There were no statistics significance between negative control group (6.04％) and blank control group (1.05％).Conclusion The siRNA in vitro transcription can reduce the level of GRP78 and GRP94 in SGC-7901 efficiently,and can inhibit the proliferation and increase the ratio of apoptosis of SGC-7901 in vitrv. Key words: RNA interference; Glucose regulative proteins 78 ; Glucose regulative proteins 94 ; Gastric cancer cells ; Apoptosis