| Objectives:1.To detect the effects of monotherapy and combined treatment of scutellarin and gemcitabine on human pancreatic cancer PANC-1 cells on cell proliferation,cell morphology.Moreover,investigate whether combined treatment of scutellarin and gemcitabine could synergistically inhibit cell proliferation.2.To detect the effects of monotherapy and combined treatment of scutellarin and gemcitabine on human pancreatic cancer PANC-1 cells on cell viability,cell apoptosis rate.Moreover,investigate whether combined treatment of scutellarin and gemcitabine could synergistically promote apoptosis.3.The possible functional mechanism of scutellarin and gemcitabine monotherapy and combined treatment on human pancreatic cancer PANC-1 cells was explored,and the changes of related gene expression in this process were detected.Methods:1.CCK-8 assay was used to determine the effects of scutellarin and gemcitabine monotherapy and combined treatment in a certain concentration and time range on the inhibition rate of human pancreatic cancer PANC-1 cells in vitro.2.The effects of scutellarin and gemcitabine on human pancreatic cancer PANC-1cells on cell morphology were observed by inverted microscope.3.The effects on human pancreatic cancer PANC-1 cells on cell viability was determined by trypan blue staining.4.The effects on human pancreatic cancer PANC-1 cells on apoptosis rate was detected by PI double staining flow cytometry.5.QPCR was used to detect the effects of scutellarin and gemcitabine on the expression of P53,Bcl-2 and Bax related genes in human pancreatic cancer PANC-1cells.Results:1.Effects of scutellarin and gemcitabine monotherapy and combined treatment on cell inhibition rate in vitro at a certain concentration and time range?1?The cell inhibition rates by scutellarin with different concentrations(200.00,240.00,280.00,320.00?g.mL-1)showed significant time-and concentration-dependent effects after 24,48,72 h.The higher the scutellarin concentration,the lower the cell inhibition rate;and the longer the scutellarin treatment time,the higher the cell inhibition rate.?2?The cell inhibition rates by gemcitabine with different gemcitabine concentrations(81.38?162.76?244.14?325.52 nmol.L-1)showed no significant correlation between the cell inhibition rate and the concentration after 24,48,72 h.At the specific concentration(81.38 nmol.L-1),the cell showed the highest inhibition rate,but there was a certain correlation between the cell inhibition rate and the treatment time.The longer the treatment time,the higher cell inhibition rate.?3?After 72 h of scutellarin and gemcitabine monotherapy and combined treatment,the cell inhibition rates of scutellarin group,gemcitabine group and the combined treatment group were all increased,and the inhibition rate of the combined treatment group were significantly higher than that of the monotherapy group,indicating that the combined treatment of scutellarin and gemcitabine had a synergistic inhibitory effect on cell proliferation.2.Effects of scutellarin and gemcitabine monotherapy and combined treatment on cell morphologyThe human pancreatic cancer PANC-1 cells were cultured for 24 h in 4 different ways,and then observed their cell morphology under an inverted microscope:normal cells were adherent,mostly were epithelioid polygons,have uniform cell size,integrated cell membrane and clear nucleus.After treatment with scutellarin and gemcitabine,the cells showed morphological changes:the cells became larger and rounder,the cell membrane was incomplete,the intercellular space became larger,and the cells were even detached in some severe cases.The morphological changes mentioned above were more obvious when scutellarin and gemcitabine combined treatment.This indicated that scutellarin combined with gemcitabine could synergistically promote apoptosis of pancreatic cancer PANC-1 cells.3.Effects of scutellarin and gemcitabine monotherapy and combined treatment on cell vitalityThe human pancreatic cancer PANC-1 cells were cultured for 24 h in 4 different ways,compared with the negative control group,the survival rate of cells in scutellarin group(200.00?g.mL-1),gemcitabine group(81.38 nmol.L-1),and the combined treatment group were decreased,moreover,the survival rate of cells in combined treatment group decreased more significantly than that in the monotherapy group.This indicated that scutellarin and gemcitabine combined treatment could synergistically suppress the proliferation of pancreatic cancer PANC-1 cells.4.Effects of scutellarin and gemcitabine monotherapy and combined treatment on apoptosis rateThe human pancreatic cancer PANC-1 cells were cultured for 24 h in 4 different ways,compared with the negative control group,the apoptosis rate of cells in scutellarin group(200.00?g.mL-1),gemcitabine group(81.38 nmol.L-1),and the combined treatment group were increased,and the apoptosis rate in combined treatment group increased more obviously than that in monotherapy group.This indicated that scutellarin and gemcitabine combined treatment could synergistically promote apoptosis of pancreatic cancer PANC-1 cells.5.Effects of scutellarin and gemcitabine monotherapy and combined treatment on the expression of P53,Bcl-2,Bax related genes?1?The human pancreatic cancer PANC-1 cells were cultured for 24 h in 4different ways,there was no evident difference in the relative expression of P53 gene between the scutellarin and gemcitabine monotherapy and combined treatment group.?2?The human pancreatic cancer PANC-1 cells were cultured for 24 h in 4different ways,compared with the negative control group,the relative expression of Bcl-2 in scutellarin group(200.00?g.mL-1),gemcitabine group(81.38 nmol.L-1),and the combined treatment group were all downregulated,and the difference was statistically significant when compared with the negative control group.?3?The human pancreatic cancer PANC-1 cells were cultured for 24 h in 4different ways,compared with the negative control group,the relative expression of Bax in scutellarin group(200.00?g.mL-1),gemcitabine group(81.38 nmol.L-1),and the combined treatment group were all increased,and the difference was statistically significant when compared with the negative control group.Conclusion:1.Scutellarin and gemcitabine monotherapy treatment had inhibitory effects on cell proliferation,and the inhibitory effects was more significant when these two medicines combined treatment.2.Scutellarin and gemcitabine monotherapy treatment had promotion effects on cells apoptosis,and the promotion effects was more significant when these two medicines combined treatment.3.The anti-pancreatic cancer effect of scutellarin might mediated by cell apoptosis through up-regulating the relative expression of Bax genes and down-regulating the Bcl-2 gene expression of induce apoptosis. |
PDF Full Text Request