Study On Apoptosis Of BGC-823 Cells Induced By Soluble Human TRAIL

Objective: Constructing a plasmid of pBud-EGFP-TRAIL to study the effect of soluble TRAIL protein (sTRAIL)on the gastric cancer cell line of BGC-823 in vitro and vivo.Methods: EGFP was as the report gene while sTRAIL as the target gene, to form a recombinated plasmid of pBud-EGFP-TRAIL,which was then identified by the techonology of restriction digest and sequencing. After it had been transfected into gastric cancer cells of BGC-823 by LipofectamineTM2000, the expression of EGFP was observed by fluorescent microscope , RT-PCR and Western Blot were used to detected the expression of sTRAIL. The inhibition of cell proliferation was examined by MTT. The method of TUNEL and flow cytometry were selected to detect apoptosis. Cells of BGC-823 were injected into nude mice hypodermically to form a transplantation tumor, and the plasmid of pBud-EGFP-TRAIL was then injected into the tumors. The changes of sTRAIL were detected by RT-PCR and Western blot. The size of tumors were compared between the experimental group and the control ones. H.E dyeing and TUNEL were used to detect the pathological change and apoptosis in the tumors. Results: Sequencing result showed that the recombinant plasmid of pBud-EGFP-TRAIL had been constructed successfully. After the transfection of it into the BGC-823 cells, sTRAIL protein was expressed and cells were induced to apoptosis. The inhibition rate was higher than that of the control ones though MTT, and the apoptosis rate was detected higher through the flow cytometry. Karyopycnosis, nuclear chromosomal condensation and segmentation were observed by TUNEL. In vivo, the results showed that the volume of tumors in the pBud-EGFP-TRAIL group is much smaller than that of the control ones, and more cell death and apoptosis were detected through H.E dyeing and TUNEL.Conclusion: It has been initially proved that the recombinated eukaryotic expressing plasmid of pBud-EGFP-TRAIL can induce apoptosis of gastric cancer cell of BGC-823 both in vitro and vivo ,which is a basic foundation for the genetic therapy of gastric cancer in future.