AIM: To investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng,5-fluorouracil and adriamycin,and the possible molecular mechanisms of delisheng...AIM: To investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng,5-fluorouracil and adriamycin,and the possible molecular mechanisms of delisheng. METHODS: Human hepatocellular carcinoma HepG2 cells were cultured with a liquid overlay technique. After the formation of multicellular spheroids,morphology was analyzed by phase contrast microscopy,scanning electron microscopy and transmission electron microscopy. Sensitivity of HepG2 cells to delisheng,5-fluorouracil and adriamycin was investigated by MTT assay in multicelluar spheroids and monolayers. Vascular endothelial growth factor (VEGF) and endostatin expression were analyzed in multicellular spheroids treated with delisheng,5-fluorouracil,adriamycin and negative control PBS,with immunohistochemical staining. RESULTS: Multicellular spheroids exhibited structural characteristics somewhat different to those in monolayers. The cells in three-dimensional cell culture turned out to be less sensitive to delisheng,5-fluorouracil and adriamycin than the cells cultured in monolayer. This showed that delisheng had a satisfactory cells inhibition ratio compared to 5-fluorouracil and adriamycin. Immunohistochemical staining showed that VEGF and endostatin expression was positive during growth as multicellular spheroids,and endostatin expression in spheroids with treatment of delisheng was higher than that with 5-fluorouracil,adriamycin and PBS (139.35 ± 7.83,159.23 ± 10.34,162.83 ± 3.47 and 148.48 ± 11.06,P < 0.05).CONCLUSION: Chinese medicine compound delisheng has satisfactory anti-tumor activity in HepG2 cells in three-dimensional culture,and the effects are associated with up-regulation of endostatin.展开更多
Objective:To investigate the effect of Delisheng Injection（得力生注射液 DLS）,a Chinese medicinal compound,DLS combined with cis-platinum（DDP）,an active agent used in lung cancer chemotherapy,on a human highly meta...Objective:To investigate the effect of Delisheng Injection（得力生注射液 DLS）,a Chinese medicinal compound,DLS combined with cis-platinum（DDP）,an active agent used in lung cancer chemotherapy,on a human highly metastatic giant lung carcinoma cell line PGCL3.Methods:The suspended PGCL3 cells at10;/mL cultured in 96-well tissue culture plates were divided into 4 groups:DLS treatment group（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）,DDP treatment group（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL）,combined DLS with DDP treatment group（DLS:DDP 2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL）and a control group.The cytotoxicity of DLS with different concentrations（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）on PGCL3 cells was determined by 3-（4,5）-dimethylthiahiazo（-z-y1）-3,5-di-phenytetrazoliumromide（MTT） assay.Effect of DLS on adhesion of PGCL-3 cells was tested by cell-matrigel adhesion assay.Chemotactic movement model of transwell camerula was used to determine the effect of DLS on invasion and migration of PGCL-3 cells.Results:Compared with the control group,DLS（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL） could significantly decrease cell proliferation,adhesion,invasion and migration abilities（P<0.05）.Cell adhesion,invasion and migration abilities were significantly decreased after combination treatment of DLS:DDP（2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL） compared with DDP single-agent treatment（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL,P<0.05）,respectively.Conclusions:DLS single-agent has a satisfying inhibition effect in PGCL3 cell line and DLS might enhance the inhibition effect of DDP on cancer metastasis.Our research provided a experimental basis about the treatment on highly metastatic lung caner.展开更多
文摘AIM: To investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in three-dimensional culture to delisheng,5-fluorouracil and adriamycin,and the possible molecular mechanisms of delisheng. METHODS: Human hepatocellular carcinoma HepG2 cells were cultured with a liquid overlay technique. After the formation of multicellular spheroids,morphology was analyzed by phase contrast microscopy,scanning electron microscopy and transmission electron microscopy. Sensitivity of HepG2 cells to delisheng,5-fluorouracil and adriamycin was investigated by MTT assay in multicelluar spheroids and monolayers. Vascular endothelial growth factor (VEGF) and endostatin expression were analyzed in multicellular spheroids treated with delisheng,5-fluorouracil,adriamycin and negative control PBS,with immunohistochemical staining. RESULTS: Multicellular spheroids exhibited structural characteristics somewhat different to those in monolayers. The cells in three-dimensional cell culture turned out to be less sensitive to delisheng,5-fluorouracil and adriamycin than the cells cultured in monolayer. This showed that delisheng had a satisfactory cells inhibition ratio compared to 5-fluorouracil and adriamycin. Immunohistochemical staining showed that VEGF and endostatin expression was positive during growth as multicellular spheroids,and endostatin expression in spheroids with treatment of delisheng was higher than that with 5-fluorouracil,adriamycin and PBS (139.35 ± 7.83,159.23 ± 10.34,162.83 ± 3.47 and 148.48 ± 11.06,P < 0.05).CONCLUSION: Chinese medicine compound delisheng has satisfactory anti-tumor activity in HepG2 cells in three-dimensional culture,and the effects are associated with up-regulation of endostatin.
基金Supported by the Project of Science and Technology Development in Shaanxi Province of China(No.2008KG10-01)
文摘Objective:To investigate the effect of Delisheng Injection（得力生注射液 DLS）,a Chinese medicinal compound,DLS combined with cis-platinum（DDP）,an active agent used in lung cancer chemotherapy,on a human highly metastatic giant lung carcinoma cell line PGCL3.Methods:The suspended PGCL3 cells at10;/mL cultured in 96-well tissue culture plates were divided into 4 groups:DLS treatment group（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）,DDP treatment group（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL）,combined DLS with DDP treatment group（DLS:DDP 2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL）and a control group.The cytotoxicity of DLS with different concentrations（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）on PGCL3 cells was determined by 3-（4,5）-dimethylthiahiazo（-z-y1）-3,5-di-phenytetrazoliumromide（MTT） assay.Effect of DLS on adhesion of PGCL-3 cells was tested by cell-matrigel adhesion assay.Chemotactic movement model of transwell camerula was used to determine the effect of DLS on invasion and migration of PGCL-3 cells.Results:Compared with the control group,DLS（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL） could significantly decrease cell proliferation,adhesion,invasion and migration abilities（P<0.05）.Cell adhesion,invasion and migration abilities were significantly decreased after combination treatment of DLS:DDP（2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL） compared with DDP single-agent treatment（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL,P<0.05）,respectively.Conclusions:DLS single-agent has a satisfying inhibition effect in PGCL3 cell line and DLS might enhance the inhibition effect of DDP on cancer metastasis.Our research provided a experimental basis about the treatment on highly metastatic lung caner.
