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Objective To investigate the effect of human platelet-rich plasma-derived exosomes(PRP-exos)on the proliferation of Schwann cell(SC)cultured in vitro. Methods PRP-exos were extracted by polymerization-precipitation combined with ultracentrifugation.The morphology of PRP-exos was observed by transmission electron microscopy,and the concentration and particle size distribution of PRP-exos were determined by nanoparticle tracking analysis.Western blotting was employed to determine the expression of the marker proteins CD63,CD81,and CD9 on exosome surface and the platelet membrane glycoprotein CD41.The SCs of rats were isolated and cultured,and the expression of the SC marker S100β was detected by immunofluorescence staining.The fluorescently labeled PRP-exos were co-cultured with SCs in vitro for observation of their interaction.EdU assay was employed to detect the effect of PRP-exos on SC proliferation,and CCK-8 assay to detect the effects of PRP-exos at different concentrations(0,10,20,40,80,and 160 μg/ml)on SC proliferation. Results The extracted PRP-exos appeared as uniform saucer-shaped vesicles with the average particle size of(122.8±38.7)nm and the concentration of 3.5×1012 particles/ml.CD63,CD81,CD9,and CD41 were highly expressed on PRP-exos surface(P<0.001,P=0.025,P=0.004,and P=0.032).The isolated SCs expressed S100β,and PRP-exos could be taken up by SCs.PRP-exos of 40,80,and 160 μg/ml promoted the proliferation of SCs,and that of 40 μg/ml showed the best performance(all P<0.01). Conclusions High concentrations of PRP-exos can be extracted from PRP.PRP-exos can be taken up by SCs and promote the proliferation of SCs cultured in vitro.
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Humanos , Ratos , Animais , Exossomos/metabolismo , Plasma Rico em Plaquetas , Células de Schwann , Técnicas de Cocultura , Proliferação de Células , Células CultivadasRESUMO
Objective: To investigate the expression of circular RNA circSP3 (hsa-circ-0002642) in hepatocellular carcinoma (HCC) and its effect on proliferation and migration of HCC cells. Methods: The tumor tissue and the adjacent paratumor tissue samples were collected from 42 HCC patients who underwent surgery from Jun. 2017 to Dec. 2018 in Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University. The expression of circSP3 in the samples was detected by real-time quantitative PCR, and the relationship between circSP3 expression in the tumor tissues and clinicopathological parameters of the patients was analyzed. Human HCC cell lines (Hep-3B, Huh7, SMMC-7721 and Bel-7402) and human normal liver cell line (HL-7702) culturion, and the expression of circSP3 was detected. After circSP3 overexpression and interference plasmids transfection into Hep-3B and Huh7 cells, the cell proliferation was detected by cell counting kit-8 (CCK-8) method, the invasion and migration abilities were detected by Transwell assay, and expression levels of epithelial-mesenchymal transformation (EMT)-associated markers (vimentin and E-cadherin) were determined by Western blotting. Results: The expression of circSP3 of tumor tissues was significantly higher than that of the paratumor tissues in the HCC patients (P < 0.01), and the expression of circSP3 was positively correlated with the tumor maximum diameter and clinical TNM stage (both P < 0.05). The expression levels of circSP3 in the 4 HCC cell lines were significantly higher than that in the normal liver cell lines (all P < 0.01). Overexpression of circSP3 could significantly promote proliferation, invasion and migration of HCC cells (P < 0.05, P < 0.01), while interference circSP3 expression could significantly inhibit the proliferation, invasion and migration (P < 0.05, P < 0.01). The expression of vimentin was significantly higher in circSP3-overexpressed cells than that in control cells (P < 0.05), while it was significantly lower in circSP3-interfered cells than that in control cells (P < 0.05). The expression of E-cadherin was significantly lower in circSP3-overexpressed cells than that in control cells (P < 0.01), while it was significantly higher in circSP3-interfered cells than that in control cells (P < 0.01). Conclusion: The abnormal expression of circSP3 is positively correlated with tumor size and TNM stage of HCC, and determining its expression is helpful to evaluate the severity and prognosis of HCC. CircSP3 can promote the proliferation of HCC cells, and may promote the migration and invasion by promoting the EMT process.
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<p><b>OBJECTIVE</b>To investigate the toxicity attenuation and efficacy potentiation effects of Sijunzi decoction (SJZD) on bladder carcinoma treated by chemotherapy in mice.</p><p><b>METHODS</b>T739 mice were randomly divided into 8 groups after subcutaneous inoculation of bladder carcinoma cells, the control group (A); two mitomycin C (MMC) group, treated with MMC of routine dosage (B) and low-dosage (C) respectively; three SJZD groups, treated with SJZD of high (D), medium (E) and low-dosage (F) respectively; and two combined treatment groups, treated with SJZD of high-dosage + MMC of routine dosage(G) and SJZD of high-dosage + MMC of low-dosage(H). The medication was begun at 24 hrs after inoculation. The tumor inhibitory rate, activity of peritoneal macrophages after 14 days of treatment and change of peripheral white blood cells after 7 days of treatment were determined and the survival time of mice was observed.</p><p><b>RESULTS</b>The survival time of mice in Group D was significantly higher than that in Group A (P < 0.05), while those in Group E and F showed insignificant difference as compared with those in Group A (P > 0.05). The highest tumor inhibitory rate was shown in Group B, but the survival time in that group showed no significant difference as compared to those in Group A (P > 0.05). The longest survival time (32.7 +/- 1.3 days) was shown in Group H, which was obviously different to that in other groups (P < 0.05). And the leukocyte counts and macrophage activity in Group H were better than those in Group B, C and G (P < 0.05), except that the tumor inhibitory rate was significantly lower than that in Group B, C and G (P < 0.05).</p><p><b>CONCLUSION</b>Combined chemotherapy of SJZD with low dosage MMC has definite effect in inhibiting tumor growth in mice with bladder carcinoma, displaying special effects of toxicity attenuation and efficacy potentiation.</p>