ABSTRACT
Objective Based on the previous research that the ethanolic extract from traditional Chinese medicine fructus forsythiae (Lianqiao) can obviously inhibit cancer cells in vitro, the article aimed to investigate the anti-proliferation effects of dammar-24-ene-3β-acetate-20S-ol (DM) extracted from fructus forsythiae on gastric cancer cells and its mechanism.Methods MTT assay was used to assess the anti-proliferation effects of DM on gastric cancer cells including SGC-7901, BGC-823, and MKN-45 in vitro.There were MKN-45 control group and its low dose and high dose groups, BGC-823 control group and its low dose and high dose groups, SGC-7901 control group and its low dose and high dose groups in the experiment.Flow cytometry was used to analyze the cell apoptosis rate.Cellquest software was used to analyze the results and record the ratio of cells at different cycles.DCFH-DA probe was applied to detect the ROS levels of blank control group, docetaxol group and DM group.The reaction system of microtubule assembly test was set with 10?mol/L docetaxol, 50 or 100 μmol/L DM final density and no medicine in blank control group.The readings of UV spectrophotometer were recorded.Microtubule assembly assay and microtubule immunofluorescence staining were applied to investigate the effects of DM on microtubule system.Results The inhibition ratio of 50 μg/L DM on the proliferation three gastric cell lines were all above 80%, with IC50s of MKN-45 11.72±1.35 μg/mL, BGC-823 17.19±0.82 μg/mL, SGC-7901 7.55±0.79 μg/mL.8 days′ low density culturing at 48 hours after 2 μg/mL DM treatment, compared with control group, the number of cell clones significantly reduced without much change in clone size, while 48 hours after 10 μg/mL DM treatment, besides a few clones of BGC-823, there were just several megascopic clones of SGC-7901 and MKN-45.In comparison with apoptotic cell ratio in MKN-45 control group[(21.1±2.5)%], its low dose group and high dose group resulted in significant rise of apoptotic cell ratio[(25.1±1.3)% and (55.2±2.3)%] (P0.05).In comparison with MKN-45 control group, the ratio of cells at S phase decreased in its low dose group[(14.5±2.7)% vs (12.3±3.3)%,P>0.05].In comparison with BGC-823 control group, the ratio of cells at S phase increased in its low dose group[(12.2±5.4)% vs (20.2±2.1)%,P<0.05].In comparison with SGC-7901 control group, the ratio of cells at S phase increased in its low dose group[(21.5±3.8)% vs (31.3±2.6)%,P<0.05].From the detection of intracellular active oxygen after DM treatment, dose-dependent ROS level increased in all three cell lines 48 hours after 10μg/mL and 50μg/mL DM treatment.From the results of microtubule immunofluorescence staining, 48 hours after the treatment of IC50 docetaxol and 10μg/mL DM, the fluorescence signals were in local concentration and disorder.Conclusion Dammar-24-ene-3β-acetate-20S-ol demonstrated anti-proliferation effects due to the apoptosis induced by cell cycle arrest at S phase.
ABSTRACT
Aim: To investigate a new type wound dressing,basic fibroblast growth factor(bFGF)/collagen com-posite sponge,and conduct its pharmacological studies in vitro and in vivo.Methods: bFGF/collagen composite sponge was prepared using fresh pig skin and bFGF.The sponge's physicochemical properties were studied.MTT assay was used to detect the proliferation effect of the sponge extract on 3T3 cells.Delayed allergy of the sponge was tested for the assurance of its biosafety.Results: Results showed that the physicochemical properties of bFGF/collagen composite sponge with high and low doses of bFGF have no significant difference from those of blank collagen sponge.SDS-page analysis indicated that the composite sponge has apparent strip in 18 kD.It was also found that bFGF/collagen composite sponge was responsible for significant effects on 3T3 cell proliferation in comparison to saline treatement(P <0.01,P <0.05).In the allergy study,during the periods of the induction and stimulation,no allergic reaction was found in bFGF/collagen composite sponge groups with high and low doses of bFGF,while severe reactions and inflammation occurred in positive group(2,4-dinitrochlorobenzene).Furthermore,pathological examination indicated the intact dermal structure and no sign of inflammation.Conclu-sion: The developed sponge has good physicochemical propertis and noticed cellular proliferation without dermal irritation.There is much potential to develop bFGF/collagen composite sponge into a new kind of wound dressing material for clinical use.
ABSTRACT
Objective To understand the effects of 17?-estradiol on MCF-7 cell proliferation and the regulation of estrogen receptor-related recepto(rERR?).Methods MCF-7 cells were treated with 17?-E2 at the doses of 10-12,10-11,10-10,10-9,10-8,10-7 and 10-6 mol/L for 24 h.Dimethyl sulfoxide were used as the solvent control.MTT method was used to determine inhibitory rate of MCF-7 proliferation,and the cells were treated with 17?-E2 at the doses of 10-10,10-9,10-8 and 10-7 mol/L for 24 h.RT-PCR method was used to detect the expression level of ERR?.Results Cell proliferation were promoted by exposure to 17?-E2 at the doses of 10-12,10-11,10-10,10-9,10-8,10-7 and 10-6 mol/L,the proliferation rate were 112.09%,122.09%,123.42%,120.46%,124.60% and 109.23% respectively,however,cell growth was inhibited at 10-6 mol/L of 17?-E2.The expression of ERR? in MCF-7 cells was upregulated at the doses of 10-10,10-9,10-8 and 10-7 mol/L when cells were treated with 17?-E2 for 24 h.Conclusion In certain dosage range,cell proliferation and expression of ERR? can be promoted by 17?-E2,which suggests that ERR? should play an important role in the process of MCF-7 cells growth.