The Mechanism Of Genistein On Proliferation,Migration Of Cervical Cancer Hela Cell

Genistein is one of the main components in soy foods and traditional Chinese medicine,which is the main active ingredient of soybean isoflavones.There are many reported studies have demonstrated that genistein,as a kind of common phytoestrogen,exists in a variety of traditional Chinese medicine,divided into three categories,tonic drug,blood-activating drug and other classes.In the tonic drug such as semen Cuscutae,epimedium,psoralea carylifolia L,cistanche,fructus lycii,ligustrum lucidum Ait contains genistein.In the blood-activating drug such as salvia miltiorrhiza,twotooth achyranthes root,safflower,cowherb seed also contain genistein.other drug such as soybeans,puerarin has been containing genistein.As a kind of common phytoestrogen,Genistein is widely known for many beneficial functions such as anti-cancer,anti-inflammatory and antioxidant.At present,the high incidence of cervical cancer as a kind of high malignant degree,the common tumors,are harmful to the women's reproductive health,especially the young women's health.Based on clinical situation,clinical common treatment for surgical treatment,and chemical treatment and traditional Chinese medicine treatment,etc.Used alone,or combined treatment.Although surgical treatment is the most effective treatment,the surgical treatment is mainly suitable for ? under a period of early cervical cancer patients,and requires patients without severe local complications and surgical contraindications,for patients with advanced,existing transfer or intolerance to surgical treatment of patients with limitations.Radiation therapy can be used alone,and can also be associated with surgical treatment at the same time,suitable for late cancer patients and can't tolerate surgery.Radiation treatment still has its limitations,radiotherapy for cervical cancer is not very sensitive and the surrounding normal tissue toxicity,easy to produce a number of complications,and studies have shown that radiation therapy can't prolong the survival rate.Chemical treatment has side effect on cardiovascular,liver,kidney and other organs,and can produce multi-drug resistance.So looking for an antitumor drug that can effectively improve the patients survival rate and have less toxic and side effect,is one of the most important researches.At present,the research of Chinese herbal medicine as antitumor drugs has become an important development direction of cerical cancer treatment.Treatment of traditional Chinese medicine is mainly used to eliminate pathogenic factors and strengthen the body resistance.Treatment with syndrome differentiation of traditional Chinese medicine theory.adjust the body function,can improve clinical symptoms,reduce the side reaction of radiation therapy,chemotherapy,and improve the body's ability to resist infection and immunity ability before and after operation,is one of the hot spot of current research.In this study,we screened genistein from 10 kinds of Chinese medicine effective component as the research object.mainly explores the mechanism of genistein on proliferation,migration of cervical cancer Hela Cell.The research will be described in three parts as follows:Part I A variety of traditional Chinese medicine composition screeningObjective:We already know that 10 kinds of effective components of Chinese herbal medicine have effect on a wide variety of tumor cells.This study was aimed to find the obvious effective component to delve into the next.Methods:CCK-8 assay was used to determine the 10 kinds of effective components on the proliferation of HeLa cells.Results:1.10 kinds of extracts showed different inhibitory effect on Hela cells and.2.Seven kinds of extracts(genistein,icariin,cryptotanshinone,magnolol,curcumenol,oleanolic acid,tanshinone I)in high density showed potent inhibitory effect.3.The results showed that genistein has an inhibitory effect on the cell viability in a time and dose dependent manner.Only the highest concentration of genistein(100 ?M)significantly inhibits the growth.With lower genistein(12.5-50 ?M)concentrations,no influence on the cell viability was observed.Conclusions:cervical cancer is malignant tumors which have a poor cure rate because of their invasive behavior.Nowadays,Cervical cancer recurrence after the shift has high mortality rate.CCK-8 assay was used to determine the 10 kinds of effective components on the proliferation of HeLa cells,and we find the obvious effective component to delve into the next.Part II The effact of Genistein on Proliferation,Migration of cervical cancer Hela CellObjective:Through the first part,we choose genistein for further research.We study the effect of the different concentrations of genistein on proliferation,migration of cervical cancer Hela.Methods:CCK-8 assay,cell counting and colony formation assay were used to determine the genistein on the proliferation.Wound healing mobility assay,adhesion assay,cell migration and invasion assay were used to determine the genistein on the migration and invasion.Results:1.The influence of genistein concentrations and incubation times on the viability of cervical cancer Hela cells was tested by the CCK-8 assay,colony forming assay and cell counting assay.Only the highest concentration of genistein(100 ?M)significantly inhibits the growth of Hela after 24 and 48h treatment.With lower genistein(12.5-50 ?M)concentrations,no influence on the cell viability was observed.2.The data demonstrated that pre-treatment of Hela cells with genistein for 24 h significantly inhibited cell adhesion in a dose dependent manner by the Adhesion Assay.3.With genistein treatment,the migration of Hela cells was significantly reduced by the Wound Healing Mobility Assay.4.The inhibitory effects of genistein on Hela cell migration and invasion were measured by a transwell assays.Conclusions:Our results showed that genistein strongly inhibits Hela cell proliferation,adhesion,invasion and migration in time and concentration dependent manners.Understand the antitumor function of genistein,we studied the mechanism of genistein on proliferation and migration of cervical cancer Hela in order to solve this problem,we conducted in-depth study of the next step.Part ? The mechanism of Genistein on Proliferation,Migration of cervical cancer Hela CellObjective:By the second part after the apparent results of cell proliferation,migration,we need to find out the final cause of the apparent results.Therefore,we need to be further in the protein expression and gene expression level do mechanism discussed.Methods:We explore its protein expression by Western blot,and explore its mRNA expression by RT-PCR.Results:1.Genistein treatment strongly inhibits the protein level of the phosphorylation of FAK,paxillin,?-catenin and Vimentin in a concentration dependent manner.2.Genistein treatment strongly inhibits the protein level of the phosphorylation of p38 and p42/44 in a concentration dependent manner.3.Genistein treatment strongly inhibits the gene level of the FAK,paxillin,snail and twist in a concentration dependent manner.Conclusions:genistein inhibited cell proliferation,adhesion,migration and invasion through the suppression of proteins and gene from FAK-paxillin pathway.The RNA and protein molecular target therapy for regulating network provides a new possibility for the diagnosis and treatment of cervical cancer.