Effects of doxorubicin-loaded tumor-derived extracellular vesicles on cell proliferation and apoptosis of human hepatocellular carcinoma / 中华肝胆外科杂志

ABSTRACT

Objective:To investigate the effects of doxorubicin (Dox)-loaded tumor-derived extracellular vehicles (EVs) on cell proliferation and apoptosis of human hepatocellular carcinoma.Methods:The extracellular vesicles loaded with Adriamycin (EVs-Dox) were prepared by the method of directly co-incubation. The morphology of EVs-Dox was detected by transmission electron microphotometer. The diameter of EVs-Dox was determined by dynamic light scattering (DLS). Western blotting was utilized to detect the expression of CD63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was calculated by tandem mass spectrometry (LC-MS/MS). The drug release experiment in vitro was utilized to simulate the drug release of drug-loaded vesicles in vivo. PKH67-labeled EVs-Dox was showed cellular uptake. After treatment with EVs-Dox, MTS assay and flow cytometry assay were conducted to investigate the effects of EVs-Dox on cell proliferation and apoptosis of PLC/PRF/5.Results:The EVs-Dox showed an elliptical double-layer membrane structure of different sizes under transmission electron microscope. The diameter of EVs-Dox was (115.9±5.2) nm.Western blotting data showed high expression of CD 63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was 0.77%. The in vitro release experiment showed that the drug-loaded vesicles could release the drug slowly. PKH67-labeled EVs-Dox showed that carcinoma cells can uptake EVs-Dox within 16h. MTS assay showed that the cell viability rate of (54.9±3.2) % was significantly lower than that of in the Dox group [(77.7±5.4)%, P<0.05]. EVs-Dox inhibited hepatocellular carcinoma proliferation. Flow cytometry assay showed that the apoptosis rate of EVs-Dox (47.9±7.0) % was higher than that in the Dox group [(38.0±1.5)%, P<0.05]. Conclusion:EVs-Dox inhibits cell proliferation and accelerates apoptosis of hepatocellular carcinoma cells.