Effects Of Paclitaxel On The Regulation Of Stem Genes Expression And Malignant Behavior In Liver Cancer Cells

Objective: The main stem gene markers of liver cancer include AFP,Sox2,C-myc,Klf4,EpCAM,Oct4,etc.Paclitaxel drugs can aggregate a protein in microtubules after acting on cells,thus destroying the spiral structure therein,significantly inhibiting the depolymerization of microtubules,and achieving the effect of inhibiting tumor cells.At the same time,taxanes can play a sensitizing role at higher concentrations,strengthening the polymerization of microtubules,which results in serious inhibition of depolymerization of microtubules.Thefinal effect is to inhibit the proliferation of tumor cells and then induce the death of tumor cells.In this study,the effect of paclitaxel on the expression of stem genes in liver cancer was observed under the intervention of paclitaxelin liver cancer cells,so as to explore the effect of paclitaxel on malignant behavior of liver cancer.Methods: MTT method was used to observe the growth of L-02,HLE and Bel7402 cells after 24 hours and 48 hours of intervention with different concentrations of paclitaxel.The optimal concentration and time of paclitaxel were selected.The effect of paclitaxel on the cell migration ability of L-02,HLE and Bel7402 was observed through Transwell cell migration experiment.Western blotting was used to observe the expression changes of paclitaxel on the stem genes AFP,Sox2,c-Myc,Klf4,EpCAM,Oct4 and other proteins in L-02,HLE,Bel7402 cells.DAPI staining was used to observe the morphological effects of paclitaxel on the nuclei of normal hepatocytes and hepatoma cells.We can observe the apoptosis effectof paclitaxel on normal hepatocytes and hepatoma cells through the flow cytometry.Results: The results of MTT assay showed that the cell proliferation rate of paclitaxel was significantly lower at 20 μg/mL.Western blot was used to detect the expression of stem genes AFP,Sox2,c-Myc,Klf4,EpCAM,Oct4 and other proteins in L-02,HLE and Bel7402 cells.The results showed that the concentration of paclitaxel was compared with cells without paclitaxel.The expression levels of stem genes such as AFP,Sox2,c-Myc,Klf4,EpCAM and Oct4 were significantly lower in L-02,HLE and Bel7402 cells cultured for 24 h at 20 μg/mL paclitaxel DMEM.The Transwell chamber migration test results showed that paclitaxel-acting cells were worn.The number of cells in the Transwell chamber membrane was much lower than that in the cells without paclitaxel,indicating that paclitaxel can inhibit the migration of hepatoma cells in a certain extent;DAPI staining results show that hepatoma cells under the action of paclitaxel,the nucleus is bright blue with strong refraction,dense nucleus,irregular shape,and most of the nucleus is half-moon or round,even apoptotic bodies are visible;flow cytometry results show Under the action of paclitaxel,the early apoptosis and late apoptotic cells of liver cancer cells were significantly more than those without paclitaxel,P<0.001,which was statistically significant.Conclusion: The results show that paclitaxel can reduce the expression of stem genes,include AFP,Sox2,c-Myc,Klf4,EpCAM,Oct4 and other proteins in liver cancer cells,thereby paclitaxel can inhibit the malignant behavior of liver cancer.