Clinical Microbiology: Open Access

Exosomes derived from human umbilical cord mesenchymal stromal cells deliver exogenous miR- 145-5p to inhibit pancreatic ductal adenocarcinoma progression

Joint Event on 25^th Asia Pacific Biotechnology Congress & 3^rd International Conference on Medical and Clinical Microbiology

May 01-02, 2019 Kyoto, Japan

Fei Li and Yixuan Ding

Capital Medical University, China

Posters & Accepted Abstracts: Clin Microbiol

Abstract:

Objective: To investigate whether huc-MSCs-exo could inhibit pancreatic ductal adenocarcinoma progression.

Method: Exosomes were loaded with miR-145-5p mimics and negative control miRNA by transient transfection. The proliferative activity and invasion abilities of PDAC cells were measured by CCK-8 and transwell assays. The cell cycle and cell apoptosis were detected by Flow cytometry analysis. The expression levels of SMAD3 were detected by Western blotting. Six-week-old male BALB/c nude mice were inoculated subcutaneously on both flanks with Panc-1 cells (1×106). After seven days of tumor growth, exosomes alone, NC-exo and 145-exo suspension were directly administered via intra-tumor injection for three days per week.

Result: CCK-8 assays and transwell assays demonstrated that 145-exo inhibits proliferation and invasiveness capability of PDAC cells. Flow cytometric analysis revealed that 145-exo caused an increase in the percentage of the cells in the G0/G1 stage of the cell cycle from 32.82±1.2% (NC-exo) to 43.45±1.3% (145-exo). Subsequently, we observed an increase in apoptotic cells among 145-exo treated PDAC cells. The SMAD3 protein expression was significantly down-regulated in PDAC cells upon addition of 145-exo. We also found that 145-exo led to up-regulation of the E-cadherin, concurrent with down-regulation of the N-cadherin. Moreover, the expression of the BAX was increased compared with a significant down-regulation of Bcl-2. Mice injected with 145-exo at seven days after subcutaneous inoculation had a much smaller tumor volume and weight. In addition, fewer Ki-67-positive and more TUNEL-positive cells were observed in the tumor sections in the 145-exo group.

Conclusion: 145-exo inhibit PDAC cell proliferation and invasion and increased apoptosis and cell cycle arrest, concomitant with decreased SMAD3 expression in vitro. We also demonstrated that 145-exo significantly reduced the growth of xenograft tumors in vivo.

Biography :

E-mail: lifei20180619@163.com