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Chinese Journal of Hepatobiliary Surgery ; (12): 247-252, 2020.
Article in Chinese | WPRIM | ID: wpr-868808


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.

Chinese Journal of Tissue Engineering Research ; (53): 4491-4497, 2014.
Article in Chinese | WPRIM | ID: wpr-452957


BACKGROUND:The obesity has led to a plenty of diseases including hypertension, coronary heart disease, fatty liver, hyperlipidemia, and type 2 diabetes. Therefore, understanding the mechanism of adipocyte differentiation is of far-reaching significance to the prevention and treatment of obesity. For the current studies of the mechanism of adipocyte differentiation pay more attention to microRNA, rather than long non-coding RNAs (lncRNAs). OBJECTIVE:To obtain the lncRNAs whose fold change was apparent during adipogenic differentiation, and to further screen the lncRNAs that possibly play a crucial role in adipogenic differentiation for verification. METHODS:Subcutaneous fat was obtained from human abdomen. Adipose-derived stem cells were col ected using tissue culture method. The third passage of adipose-derived stem cells was used for adipogenic differentiation. Through microarray technology, the expression levels of lncRNAs and mRNA were analyzed at 0, 5 and 12 days in adipogenic differentiation. Combining with bioinformatics report, lncRNAs apparently presented fold change were screened and verified by qRT-PCR. RESULTS AND CONCLUSION:Fold change 1.5 (P<0.05) was considered as a criterion during adipogenic differentiation. The number of up-regulated lncRNAs was 748 for 5 days versus 0 day, 847 for 12 days versus 0 days, 593 for 12 days versus 5 days. At the same time, the down-regulated number was 828 for 5 days versus 0 day, 1 113 for 12 days versus 0 day, 750 for 12 days versus 5 days during adipogenic differentiation. In combination with bioinformatics analysis results, 3 of 28 lncRNAs were related to lipid metabolism:AK304548, BP216319 and DA852857, according to the standard that fold change in 0, 5 and 12 days was higher, and the target genes were known to be associated with adipogenesis-related genes. PCR results showed that the expression of AK304548 and BP216319 and its target gene presented an up-down trend, which is consistent with the microarray sequencing results. These results indicated that lncRNA plays a critical regulatory role in the adipogenic differentiation.