| [Background and objective]Polydatin(PD)is a kind of monomer that obtained from the dried rhizome of cuspidatum,and it is also known as cuspidatum crystal 4th.In addition,there are a lot of resveratrol in grapes and red wine.In recent years,some studies about resveratrol at home and abroad have shown that resveratrol has a significant effect in antiplatelet aggregation,anti-thrombosis,enhancing cardiomyocyte contraction and diastolic function,and improving microcirculation of important tissues and organs after patients are in a shock,In addition,Polydatin also play an important role on the lung injury of anti-endotoxin shock,protecting liver cells,lipid-lowering,anti-lipid peroxidation,reducing chronic pain caused by endometriosis and improving the symptom of colitis.Apoptosis is a phenomenon that occurs mostly in the physiological condition of cell death,so it is named physiological death,apoptosis,also known as cell programmed death.The important feature of the cells is the swell,dissolution,and the releasing of lysis products of cells,which making the surrounding tissue produce inflammatory response.In 1972,Kerr proposed this concept first.Apoptosis is an important self-stabilizing mechanism for multicellular organisms.It can remove the excess cells initiatively,which is specially,of which the differentiation ability is not compatible with the body,which have completed its task and is useless,which is potentially dangerous,such as the own reactive lymphocytes and the cells with the damaged DNA that cannot be repaired that are at risk of cancer.Therefore,apoptosis is an essential mechanism for regulating the organisms'normal development,and the failure of regulation will lead patients to suffer from some disease,such as cancer and even death.Tumor cells are immortal,and the occurrence of tumor are not only related with the cells' abnormal proliferation and differentiation,but also related with the cells' abnormal apoptosis.Polydatin is the derivative of resveratrol,and a large number of clinical studies have shown that resveratrol has a good effect on preventing tumor and curing cancer.However,the studies that whether Polydatin has a good effect on prevention and treatment of cancer have just started.It has been observed that Polydatin has effects on transplanted hepatocellular carcinoma in mice and Ehrlich ascites carcinoma has been found,and can inhibit tumor from growing continuous and arrest cell cycle in G2/M phase.The aim of this study was to investigate whether resveratrol could inhibit the proliferation of hepatoma cells in vitro and in vivo,and to further understand the effect of resveratrol on the apoptosis and motility of human hepatoma cell line HepG2.The study provides a broader experimental basis.The aim of our study is to provide the broader experimental basis for further understanding of the mechanism of the effect of Polydatin on the apoptosis of human hepatocellular carcinoma cell and its mechanism.This topic will be studied around the relevant issues.[Methods]In this study,we use human hepatocellular carcinoma cell lines HepG2 and use the CCK-8 to detect the cell viability,and we use it to detect the effects on human hepatocellular carcinoma cell after treat them with different concentrations(12.5?200?mol/L)at different time(24,48,72h).The cell and nuclear changes were observed by inverted microscope and fluorescence microscope.Flow cytometry was used to detect the apoptosis rate of human hepatocellular carcinoma cells after the treatment.The effect of resveratrol on the invasion ability of hepatocellular carcinoma cells was detected by adhesion test,exercise test and invasion assay.The effects of different cell lines on the cell invasion were analyzed.The results were expressed as(mean± SD),using the variance analysis of factorial design and single factor analysis of variance.The LSD method was used to compare the mean numbers.When it was not homogeneity of variance,we use the Dunnet-t method.And we use SPSS 19.0 statistical software to process the data.[Results]1.Polydatin inhibited the growth of hepatocellular carcinoma cellsThe results of CCK-8 experiment showed that after treating the cells in 24-72h with different concentration of Polydatin that was in the range of 12.5?200?mol/L,we found that the proliferation of HepG2 cell line was inhibited with concentration-dependent manner and time-dependent manner(P<0.05).After we treated the cells with Polydatin that was above 12.5?mol/L,we found that the proliferation of HepG2 cells was inhibited.And when it reached 25?mol/L,the apoptosis of HepG2 cells was induced.2.Polydatin induced apoptosis of hepatocellular carcinoma cellsFlow cytometry analysis showed that when the cells were treated with different concentrations of Polydatin,and the cells were detected by flow cytometry at different time points.Compared with the control group,the percentage of apoptosis was measured with the concentration and time.It can be seen that Polydatin can induce apoptosis of cells in a time-dependent manner.3.Polydatin changed the morphology of hepatocarcinoma cells and nucleiInverted microscope observation:morphological changes of 24h group has been seen obvious differences.The cells in the control group was adherent as big,fusiform or polygonal with good light transmittance and high cell density.Only a small amount of cells were shedding in the culture medium.The cell growth status of the experimental group was significantly worse than that of the control group.The cells were shrunk.With the increase of the concentration,the cells survived less and the floating cells increased.The same concentration group was significantly different in different time groupsHoechhest33342 fluorescence staining showed that the cell and nucleus morphology changed significantly after treatment with Polydatin,cell volume was reduced and deformed,apoptotic bodies were formed,nucleus nuclear was pyknotic,nuclear fragmentation and nuclear dissolution were observed,when cells in the apoptotic stage,the nucleus became significantly different with the normal ones with the concentration and time-dependent manner.4.Polydatin reduced the invasive ability of hepatocarcinoma cellsThe results showed that the ability of the invasion and metastasis of hepatocellular carcinoma cells were significantly decreased with the increase of the concentration of polydatin and the time of treatment.At the concentration of Polydatin in 25?mol/L and 50?mol/L,it could inhibit the adhesion and the invasive ability of HepG2 cells(P<0.01).[Conclusion]Polydatin mainly inhibited the proliferation of hepatocarcinoma cells and induced the apoptosis of hepatocellular carcinoma cells in the concentration-dependent and time-dependent manner,and it inhibited the invasive ability of hepatocarcinoma cells though inhibiting the adhesion of hepatocellular carcinoma cells between FN and the degradation of the basement membrane matrix,and it also can reduced the invasive ability and metastatic ability of hepatocellular carcinoma cells. |
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