半夏泻心汤对胃癌细胞来源外泌体诱导BMSCs体内致瘤的影响OA

Objective To observe the tumorigenicity of bone marrow mesenchymal stem cells(BMSCs)induced by gastric cancer cell-derived exosomes and the effect of Banxia Xiexin decoction treatment.Methods Exosomes were isolated from human gastric cancer NCI-N87 cells,characterized by transmission electron microscopy and Western Blot,and rat BMSCs were passaged and characterized by inverted microscope and immunofluorescence staining.Sixty SPF BALB/c-Nude mice were randomly divided into and control,model,and high-,medium-,low-dose Banxia Xiexin decoction and groups(n=10 per group).The control group was inoculated with normal BMSC suspension(200 μL in axilla);other groups were inoculated with the same dose of BMSC suspension induced by co-culture of exosomes from gastric cancer cells and BMSCs.High-,medium-,low-dose groups were given 1.56,0.78,0.39 g/kg Banxia Xiexin decoction by gavage,respectively,once a day(0.2 mL each time).The positive group was given 0.015 g/kg 5-FU by intraperitoneal injection once every 3 d(0.2 mL each time).The control group received 0.9％sodium chloride solution at an equal volume.Tumor volume was measured every 7 d.After 21 d,subcutaneous tumors were stripped and mass,tumor length and diameter were measured,and tumor volume and inhibition rates were calculated.The tumor tissue ultrastructure was observed by transmission electron microscopy,and latent membrane protein 1(LMP1)and transcription factor 21(TCF21)expression was detected by immunohistochemical staining.Results No tumor formation was observed in the control group after inoculation with normal BMSCs.In the remaining groups,subcutaneous tumors appeared in the axilla on day 14 after inoculation with BMSCs.Compared with the model group,the tumor volumes in the high-and medium-dose Banxia Xiexin decoction groups and the positive group were reduced(P＜0.05).On day 21,the axillary tumor volume in each group increased significantly.Compared with those in the control group,tumor volume and mass in the model group were significantly increased,while those in the positive and high-and medium-dose Banxia Xiexin decoction groups were significantly smaller than those in the model group(P＜0.05);these groups also exhibited tumor inhibition,which was most significant in the high-dose Banxia Xiexin decoction group(P＜0.05).Transmission electron microscopy showed that tumor cells in the control group were densely distributed,with long spindle-and irregular-shaped nuclei.The nuclear membrane was relatively level,and intranuclear euchromatin was abundant,with subcutaneous fibrous tissue between cells.The cells in the model group were arranged closely,with abundant organelles.The nuclei in each dose group of Banxia Xiexin decoction were irregular and damaged to different degrees.Mucus particles in the nuclei were decreased,small,and high-density,especially in the high-dose Banxia Xiexin decoction group.Immunohistochemistry showed that LMP1 protein expression was increased in the tumor tissue of the model group,while TCF21 protein expression was decreased(P＜0.05).After intervention,LMP1 protein expression was decreased in the tumor body tissue of the positive group and each dose group,while that of TCF21 was increased(P＜0.05).LMP1 and TCF21 expression changes were most significant in the high-dose Banxia Xiexin decoction group(P＜0.05).Conclusions BMSCs induced by gastric cancer cell-derived exosomes had tumorigenic properties in vivo.The mechanism involved may be related to increasing LMP1 and reducing TCF21 expression.Banxia Xiexin decoction reduced LMP1 expression and increased that of TCF21 to inhibit BMSC tumorigenesis induced by gastric cancer cell-derived exosomes in vivo.