The Effects Of Xihuang Pills On The Proliferation And Apoptosis Of SK-NEP-1 Cells And Its Mechanism

BackgroundWilms tumor also known as nephroblastoma,embryonal tumor.It was first described by Rance in 1814.Max Wilms described its features further in 1899,so Wilms tumor was also known.It is one of the most common childhood malignant solid tumors,accounting for 95% of children with kidney tumors.The annual incidence of less than 15 years old is about 7-10 / 100,000,more than 80% of patients with nephroblastoma are under the age of 5.The average age of onset is 3.5 years old and the peak incidence age is about 3 to 4 years old.The pathogenesis of Wilms tumor is a complex multi-gene regulation process.Its pathogenesis involves multiple genes.The deletion of WT1 gene plays an important role in the pathogenesis of Wilms' disease.The WT1 gene has been shown to exist only in the kidneys,gonads and heart.Related animal experiments shown that WT1 gene mutations lead to urinary and reproductive system developmental deformity,WT1 gene deletion only can cause urinary and reproductive system abnormalities.In addition,WT2 gene,as well as other chromosomal abnormalities have a significant impact on the occurrence of nephroblastoma.Initially,peritoneal nephrectomy is the only treatment of Wilms tumor.In 1960,vincristine and doxorubicin were found to be effective in treating Wilms tumor,and chemotherapeutics began to be used in the treatment of Wilms tumor.At present,transabdominal or laparoscopic surgery combined with postoperative chemotherapy is a common regimen of nephroblastoma.However,with the continuous development and use of chemotherapeutic drugs,the drug resistance of tumor cells is constantly increasing.At the same time,due to the killing of tumor cells by chemotherapeutic drugs,normal human cells are also killed to some extent.As a result,a variety of side effects have occurred.Xihuang Pills is a traditional Chinese medicine anti-cancer prescription.It has the effection of dispersing carbuncle and detoxifying,softing hard and loosing knot and so on.Studies have shown that Xihuang Pills have inhibitory effect on human colon cancer cells and tumor cells of various nodes,and it has been widely used in the treatment of breast cancer.Yet studies have examined whether it is effective on Wilms tumor.So through the method of experiment,this study explored the action of Xihuang Pills on SK-NEP-1 cells and its possible mechanism.ObjectiveThis study is aim to investigate the effects of Xihuang Pills on the proliferation and apoptosis of nephroblastoma cell line SK-NEP-1,and to explore the underlying mechanism of action.Materials and methods1.Materials: The leachate of Xihuang Pills was made and concentrated and diluted into different concentrations to fulfil the demand of the study.2.Methods: MTS was used to detected the proliferation inhibition rate of the SK-NEP-1 cells,The flow cytometry was used to detected the rate of apoptosis in SK-NEP-1 cells,and the Western blot was used to detected the relative expression levels of Bax and Bcl-2 protein.3.Statistical analysis: Statistical software SPSS21.0 was used to analysis the data of the study.The data was shown by mean ± standard deviation(±s).Pearson correlation analysis was used to analyze the association.One-way ANOVA was used to analyze the comparison among groups.LSD-t test was used to compare the two groups.P <0.05 means the difference was statistically significant.Results1.After reaction for 72 hours,the proliferation inhibition rates of SK-NEP-1 cells in all groups of Xihuang Pills were higher than the control group,and with the increase of the concentration of Xihuang Pills,the proliferation inhibition rate also increased,the difference was statistically significant(P <0.05).2.After reaction for 48 hours,the apoptosis rates of SK-NEP-1 cells in all groups of Xihuang Pills were higher than the control group,and with the increase of the concentration of Xihuang Pills,the apoptosis rate also increased,the difference was statistically significant(P <0.05).3.After reaction for 24 hours,the relative expression of Bax protein of SK-NEP-1 cells in all groups of Xihuang Pills were higher than the control group,and with the increase of the concentration of Xihuang Pills,the relative expression of Bax protein also increased,the difference was statistically significant(P <0.05).And after reaction for 72 hours,the relative expression of Bcl-2 protein rates of SK-NEP-1 cells in all the groups of Xihuang Pills were lower than the control group,and with the increase of the concentration of Xihuang Pills,the relative expression of Bcl-2 protein rates also reduced,the difference was statistically significant(P <0.05).Conclusions1.Xihuang Pills can reduce the proliferation rate of SK-NEP-1 cells that is to say Xihuang Pills can inhibit the proliferation of SK-NEP-1 cells.2.Xihuang Pills can increase the apoptosis rate of SK-NEP-1 cells that is to say Xihuang Pills can promote the apoptosis of SK-NEP-1 cells.3.It seems that Xihuang Pills can increase the relative expression of Bax protein and reduce the relative expression of Bcl-2 protein to effect the proliferation and apoptosis of SK-NEP-1 cells.