| Background and Objective:Cervical cancer as one of the most common gynecological malignancies,its prevalence has ranked fourth in the world of women’s malignancies,mortality has also ranked fourth in the world of women’s malignancies.Cervical cancer is gradually becoming younger,and the cause of death lies in the failure of tumor treatment,recurrence and metastasis,which seriously threatens the life and health of women.Therefore,it is particularly important to explore new or adjuvant treatment methods for cervical cancer.In recent years,more and more studies have proved that flavonoids and their derivatives have anti-tumor effects and can prevent tumors,which may be related to inhibiting cell proliferation,adhesion,invasion,inducing cell differentiation,leading to cell cycle arrest,Apoptosis,etc.The total flavonoids from Baeckea frutescens are a new type of flavonoids from the analysis and purification of Baeckea frutescens,but there are few reports on its anti-tumor pharmacological effects.Therefore,the purpose of this study is to investigate effect and pharmacological of total flavonoids from Baeckea frutescens on migration,invasion and epithelial-mesenchymal transition of Cervical cancer Si Ha cells.Methods:This study was done through in vitro cell experiments.1.CCK8 kit was used to detect the proliferation and half inhibitory concentration(IC50)of cervical cancer SIHA cells in vitro by intervening five concentrations of total flavonoids from Baeckea frutescens（25ug/m L、50ug/m L、100ug/m L、200ug/m L、400ug/m L）.2.The change of in vitro migration ability of cervical cancer Si Ha cells was observed by scratch test and transfer well chamber transfer test.3.Transwell invasion assay was used to observe the changes of invasiveness of SIHA cells in vitro.4.Flow cytometry was used to detect the effect of total flavonoids from Baeckea frutescens on apoptosis of SIHA cells in vitro.5.Laser confocal microscope to observe total flavonoids from Baeckea frutescens intervention changes in the form of Si Ha cells in vitro.6.The expression of EMT-related proteins before and after the treatment with total flavonoids from Baeckea frutescens was detected by Western Blot.Result:1.Total flavonoids from Baeckea frutescens can significantly inhibit the proliferation of cervical cancer Si Ha cells.With the increase of concentration and time,the inhibition effect becomes more significant.2.Total flavonoids from Baeckea frutescens can significantly inhibit the migration ability of cervical cancer SIHA cells in vitro;In scratch test,there were statistically significant differences in 48h cell healing ability between the control group and the three drug concentration groups（P=0.00<0.001）;In Transwell chamber migration test,there were statistically significant differences in 48h cell penetration numbers between the control group and the three drug concentration groups（P=0.00<0.001）.3.Total flavonoids from Baeckea frutescens can significantly inhibit the invasion ability of Si Ha cells in vitro.In Transwell chamber invasion test,there were statistically significant differences in 48h cell penetration numbers between the control group and the three drug concentration groups（P=0.00<0.001）.4.Total flavonoids from Baeckea frutescens can induce apoptosis of cervical cancer Si Ha cells.Flow cytometry instrument test results,compared with control group,with the increase of concentration,the apoptosis rate increased significantly;The laser confocal microscope observation showed that with the increase of concentration,more and more apoptotic cells were observed in the field of vision,and the typical apoptotic morphology,such as the fragment with different size and irregular shape of the nucleus,and the set of nuclear edges,appeared.5.Western Blot results showed that total flavonoids from Baeckea frutescens can up-regulate E-cadherin protein（P=0.00<0.05）,down-regulate N-cadherin protein（P=0.00<0.05）and Vimentin（P=0.013<0.05）protein,in a dose-dependent manner.Conclusion:1.Total flavonoids from Baeckea frutescens have obvious inhibitory effect on cervical cancer Si Ha cells,which is concentration and time dependent.2.With the increase of the concentration of total flavonoids from Baeckea frutescens,the apoptosis rate increased significantly,and the number of cells in the middle and late stages of apoptosis also increased significantly.3.Total flavonoids from Baeckea frutescens inhibited the migration and invasion of cervical cancer Siha cells,and the mechanism may be related to the up-regulation of E-cadherin protein and down-regulation of N-cadherin protein and Vimentin protein。... |
