Inhibitive Effect And Mechanism Of Brucea Javanica Oil Emulsion On SiHa Cells From Cervical Cancer

PrefaceCervical cancer is the most common female genital tract cancer,female reproductive system accounted for more than half of malignant tumors,a serious threat to women's health.The incidence of cervical cancer increased year by year in our country,and the trend was younger.HPV(Human papilloma virus) infection is one of the reasons why essential to disease.Brucea javanica oil emulsion is the extracts of Brucea javanica mature fruit,it may be able to penetrate the blood-brain barrier and reduce intracranial pressure.At the same time can also be used to treat a variety of HPV-induced warts.Studies have shown that the main active ingredient-oleic acid and linoleic acid of Brucea javanica oil emulsion has a strong anti-tumor activity.Brucea javanica oil emulsion combined with radiotherapy and/or chemotherapy can prevent the decrease in white blood cell and platelet,but also promote the formation of bone marrow stem cells to regulate the immune system,thereby enhancing the quality of patient's life.It has been used in clinical lung cancer,liver cancer,metastatic brain tumors and other adjuvant therapy for malignant tumors.However,it has not to be seen that the clinical treatment of cervical cancer report.In this study,To investigate the effectiveness mechanism of Brucea javanica oil emulsion's inhibition on the SiHa cells from cervical cancer in vitro.The results could proved the basis of laboratory and foundation of Brucea javanica oil emulsion can be used for the adjuvant treatment of clinical cancer.MethodsCell culture:Culture SiHa cells in cnventional methods.The Rate Of Inhibition Of Cells:SiHa cells are transferred to 96-well cell culture plate,adding different concentrations of drugs,to continue for 24h,72h and 48h, adding MTT and DMSO to each hole,measuring the absorbance at 550 nm by Meibiaoyi.The Observation Of Apoptotic Cells:After treatment with different concentrations of Brucea javanica oil emulsion for 48 hours.Collecting all the cells, making electron microscope specimens in conventional methods,observing morphology of apoptotic cells by TEM.Detection Of The Ladder Of Apoptosis:After treatment with different concentrations of Brucea javanica oil emulsion for 48 hours.Collecting all the SiHa cells,extracting DNA in conventional methods,using Agarose gel electrophoresis, UV-light camera observation.Detection Of Apoptosis Rate:Addding different concentrations of Brucea javanica oil emulsion,incubated for 48h,collect all the SiHa cells,detect Apoptosis rate by flow cytometry.ResultsThe Inhibition Rate of Drugs to Cell:Different concentrations of Brucea javanica oil emulsion elected on the SiHa cells for 24h,48h,72h,we tested the apparent inhibitory.Cell inhibition rate is concerned with time and drug concentration. with the extension of time and drug concentration increased,the cytotoxicity gradually increased,there is time and dose-dependent manner.Cell Morphological Changes:Morphology of apoptotic cells by TEM,the control group cells Villonodular clear,membrane,nuclear envelope are complete; different concentrations of drugs after 48h,we can see that the early chromatin condensation and Margination,late in the nuclear membrane,the nuclear apoptotic bodies appear.Observation Of Agarose Gel Electrophoresis Results:20μg/ml drug groups on agarose gel electrophoresis shows a clear ladder of apoptosis,and no fragmentation in the control group and 10μg/ml drug groups.Observation Of Apoptosis Rate:The different concentrations of Brucea javanica oil emulsion added on the SiHa cells for 48h,the cells were analyzed by Flow Cytometry.Discuss This is the first observation of Brucea javanica oil emulsion's inhibition on the SiHa cells from cervical cancer in vitro.SiHa cells have a strong inhibition,and with the extension of time and drug concentration increased,the cytotoxicity gradually increased.Tumor occurrence and cell apoptosis are related,tumor cell apoptosis induced by chemotherapeutic drugs is one of the mechanisms.This study found that the cells were added Brucea javanica oil emulsion after 48h,apoptotic bodies appeared by MEM and the apoptotic DNA ladder was formated.Proving the Brucea javanica oil emulsion can induce SiHa cells apoptosis from the perspective of Morphology and biochemistry,From Flow Cytometry detecting cell apoptosis rate,we can see that the overall rate of apoptosis gradually high.The group difference was significant between early and late apoptotic rate together with control.Further explanation of the inhibit of Brucea javanica oil emulsion on SiHa cells is through induction of apoptosis.Conclusion1,Brucea javanica oil emulsion can inhibit SiHa cervical cancer cell proliferation in vitro,there is a time and concentration dependent.2,The possible mechanisms of Brucea javanica oil emulsion on SiHa cell proliferation is inducing apoptosis.