ABSTRACT
Objective To investigate the effect of human bone marrow mesenchymal stem cells (BM-MSCs) stimulated by platelets in vitro on the metastasis of cancer cells.Methods The BM-MSCs were isolated and cultured in vitro and platelets from the peripheral blood of healthy persons were purified.The MSCs (control),platelets + MSCs,and platelets treated with culture media (CM) of SGC-7901 tumor cells + MSCs (T-platelets + MSCs) were cultured,respectively,and the MSCs and supernatants (MSCs-CM and SGC-7901-CM) were collected,respectively,after 24 hours.The expressions of markers of cancer-associated fibroblasts (CAF),such as α-SMA and Vimentin,were determined by Western-blotting.The immigration ability of BM-MSCs were analyzed by Transwell assay.The levels of P-selectin in platelets stimulated by MSCs-CM or SGC-7901-CM were detected with flow cytometry.The metastasis model of gastric cancer SGC-7901 cells was established in BALB/c nude mice by the injection of tail vein,and the tumor metastasis in vivo was observed.Results The expression levels of P-selectin in platelets stimulated by MSCs-CM ([21.37 ± 1.00] ％) or SGC-7901-CM ([31.4 ± 1.71] ％ were significantly higher than that in the control ([3.17 ± 0.40] ％,t =27.85 and 29.18,P ＜ 0.01).The expression levels of α-SMA and Vimentin in platelets + MSCs group (0.79 ± 0.08 and 0.88 ± 0.01) and T-platelets + MSCs group (0.90 ±0.06 and 0.96 ±0.04) were significantly higher than that in the control (0.64 ±0.02 and 0.75 ±0.05,t =2.96 and 6.45 forα-SMA,t =4.73 and 5.73 for Vimentin,P ＜0.01).The amounts of immigration cells in platelets + MSCs group (340.3 ±27.7) and T-platelets ± MSCs group (424.3 ± 17.6) were significantly higher than that in the control (220.7 ± 19.4,t =6.14 and 13.48,P ＜ 0.01).The in vivo experimental results showed that the metastatic foci in platelets ± MSCs group (4 ± 2) and T-platelets ± MSCs group (21 ± 4) were significantly higher than that in the control (0.33 ± 0.06,t =3.051 and 8.857,P ＜ 0.01).Conclusion Platelets promote the immigration and the enhanced tumor metastasis in vivo of BM-MSCs.
ABSTRACT
Objective To establish the HPLC fingerprints of Fructus Ligustri Lucidi.Methods HPLC-ELSD analysis was carried out with Lichrospher C18 column(4.6 mm ? 250 mm,5 ?m).The detector was Alltech ELSD 2000.The method was developed by gradient elution with methanol and water as the mobile phase.With oleanolic acid as the marker,sixteen batches of Fructus Ligustri Lucidi were analyzed with computer-aided similarity evaluation system.Results HPLC fingerprints of Fructus Ligustri Lucidi showed 14 characteristic peaks and the similarity of the fingerprints of 10 batches of samples was over 0.90,indicating the fingerprints being stable and repeatable.Conclusion The method is simple,accurate and with a good reproducibility,and can be used for the identification and quality control of Fructus Ligustri Lucidi.