The molecular mechanisms of exosomes derived from human bone marrow mesenchymal stem cells for pancreatic cancer in vivo
Objective To investigate the molecular mechanism of exosomes secreted from human bone marrow mesenchymal stem cells(BMSC)for pancreatic cancer(PC)cells.Methods The nude mouse model of PC was established using PANC-1 and AsPC-1 cell lines respectively,and each cell line was divided into 3 groups:control group[mice were injected with phosphate buffer saline(PBS)via the tail vein],treatment group(PBS suspension containing exosomes was injected into the tail vein),inhibitor group[phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway inhibitor LY294002 was added,followed by exosomes].After 8 weeks,the mice were killed,tumor inhibition rate and tumor volume in situ were measured,tumor metastasis and ascites were observed,and tumor weight and liver weight were observed.The expression of PI3K and Akt in tumor cells and the expression of Survivin,matrix metalloproteinase(MMP)-9 and CD31 were detected by Western blottingThe expression of CA199 and B7-H4 was detected by ELISA.Significant differences among groups were determined by the Student t test for 2-group comparisons and One Way ANOVA followed by post hoc analysis for multiple-group compari-sons.Results The exosomes secreted by BMSC were extracted successfully.After exosome treatment,the expression of phosphorylated PI3K(p-PI3K)and phosphorylated Akt(p-Akt)was up-regulated in the two cell lines[PANC-1(p-PI3K:8.5±2.7 vs.17.9±5.6,F=19.645,p-Akt:9.3±2.9 vs.19.4±6.2,F=26.813),AsPC-1(p-PI3K:9.7±3.2 vs.19.7±6.4,F=18.578,p-Akt:8.4±2.6 vs.18.7± 6.3,F=19.817),P＜0.05],but there was no significant differernce between the control group and the inhibitor group for the p-PI3K and p-Akt(P＞0.05).After exosome treatment,the levels of pancreatic cancer markers and CD31 in the two cell lines were lower than those in the blank control group[PANC-1(B7-H4:15.9±4.8 vs.6.8±2.3,F=23.467,CA199:17.5±5.8 vs.5.8±1.7,F=29.284,Survivin:12.4±4.0 vs.4.7±1.4,F=28.714,MMP-9:10.3±3.4 vs.3.9±1.3,F=28.927,CD31:16.9±5.6 vs.6.1±1.9 F=30.168),AsPC-1(B7-H4:14.7±4.8 vs.3.1±0.9,F=18.736,CA199:16.2±5.7 vs.3.9±1.2,F=26.948,Survivin:14.4±4.7 vs.4.8±1.5,F=29.841,MMP-9:11.3±3.8vs.4.7±1.5,F=29.798,CD3:17.1±5.7vs.5.7±1.8,F=32.864),P＜0.05],while there was no statistically significant difference between the control group and the inhibitor group for the above level(P＞0.05).Conclusion Exosomes secreted by BMSCs can inhibit the prolifera-tion,invasion and metastasis of PC by activating the PI3K/Akt signaling pathway,and inhibit the angio-genesis of PC.
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