ABSTRACT
OBJECTIVE:To study the in vitro uptake of Resibufogenin(RBG)lactic acid glycolic acid copolymer-water solu-ble vitamin E (PLGA-TPGS) in human liver cancer HepG2 cells,mouse ascites-type lymphatic metastasis of tumor HCa-F cells, and the toxicity on HepG2 cells. METHODS:RCPTN loading RBG and coumarin-6(C6)were prepared. Fluorescent inverted mi-croscope was used to observe the in vitro uptake by RCPTN HepG2,HCa-F cells. It was divided into negative control group,blank PLGA-TPGS nanoparticles(EPTN)group,5-fluorouracil solution(FS)group,RBG solution(RS)group,RBG/PLGA nanoparti-cles(RPN)group and RPTN group. WST-1 was conducted to investigate the optical density at 450 nm wavelength of HepG2 cells after 24,48,72 h incubated by FS,RS,RPN and RPTN with different final concentrations (1.25,2.5,5,10,20 μg/mL);the cell viability (CV) and half inhibitory concentration (IC50) were calculated. RESULTS:RCPTN distributed around the nucleus of HepG2,HCa-F cells. CV was decreased by RBG concentration increased in RPN group and RPTN group,and decreased by time prolonged;compared with FS group,CV in RPTN group was decreased(PFS>RPN>RPTN;IC50 incubated by RPN and RPTN for 48,72 h was obviously less than that of FS and RS(P<0.05 or P<0.01). CONCLUSIONS:RPTN can deliver RBG in-to HepG2,HCa-F cells,showing inhibition effect on HepG2 cells which is stronger than RPN,RS and FS.
ABSTRACT
OBJECTIVE:To study the inhibitory effects of sinomenine(SIN)-loaded polylactic-co-glycolic acid-D-α-tocopherol polyethylene glycol 1000 succinate(PLGA-TPGS)nanoparticles(SPTN)on the proliferation of HCa-F cells in lymph tubes and ec-topic transplantation tumors in mice. METHODS:HCa-F cell suspension were incubated with normal saline,5-fluorouracil(FS), sinomenine solutions (SS),sinomenine PLGA nanoparticles (SPN) and SPTN,with concentration of 80 μg/ml. The cells were marked with CFSE,and then were injected with suspension 50 μl via one footpad of mice. Inhibitory effect of above suspensions on the proliferation of HCa-F in lymph tubes of mice was observed by fluorescence inverted microscope at 3,6,9,12,24 h(n=15). Mice were divided into normal control group,blank PLGA-TPGS nanoparticles (EPTN) group,normal saline group,SPTN group,SPN group,SS group and FS group with 10 mice in each group. The latter 5 groups were injected with relevant medicine 15 mg/kg,once a day,via tail vein for consecutive 10 days after the model of HCa-F cells-bearing ectopic transplantation tumor mice was established. The serum content of ALT,AST,γ-glutamyltransferase(γ-GT),ALB and T-BIL were determined;solid tu-mors were taken,measured and weighed,and the inhibitory rate of tumor was also calculated. RESULTS:The inhibitory effect of above solution on the proliferation of HCa-F cells in descending order was as follows:SPTN>SPN>FS>SS>normal saline. Com-pared with normal saline group,the serum levels of ALT,AST,γ-GT and TBIL of SPTN group,SPN group,SS group and FS group decreased,while ALB level increased(P<0.05);the amount of tumor volume increase and tumor weight in SPTN group, SPN group and FS group decreased significantly (P<0.05). The inhibitory rate of tumor in 3 groups were 49.62%,40.53% and 33.90%. CONCLUSIONS:SPTN can inhibit the proliferation of HCa-F cells in lymph tubes of mice,and can improve HCa-F cells-bearing ectopic transplantation tumor in mice. It is better than SPN and FS.
ABSTRACT
Objective L-selectin is an important adhesion molecule,which is the lymph cell's homing receptor specifically expressed on leukocytes.A non-lymphoma cell line-HCa-F has the high potential ability of metastasis to peripheral lymph nodes,whose character is similar to lymphoma's.We want to explore whether the HCa-F cell's metastasis mechanism is also similar to lymphoma's,that is to say,whether HCa-F cell can also express L-selectin and the expression of L-selectin bring about the lymph node metastasis character. Methods First,?-fetoprotein,CD3 and CD20's monoclonal antibodies were used to identify the origin of HCa-F cell;then we analysed whether the HCa-F cell can express L-selectin by means of flow cytometry analysis,RT-PCR,and Western blotting;finally,Stamper-Woodruff method was used to examine the L-selectin's function. Results Immunostaining showed that the HCa-F cell was ?-fetoprotein positive,CD3 and CD20 negative cell line,which suggests that it is a liver cell line.And results of flow cytometry,RT-PCR,and Western blotting showd that HCa-F cell can express L-selectin.Additionally,anti-L-selectin's antibody can inhibit the adhesion between HCa-F cells and frozen section of mouse lymph node.Conclusion L-selectin expressed on HCa-F cells is a functional molecule which can facilitate HCa-F cell's metastasis to lymph nodes.