ABSTRACT
Objective To investigate the mechanisms of miRNA-15a and miRNA-16 in the process of reversing cisplatin resistance in ovarian cancer.Methods Human ovarian cancer cisplatin-resistant cell lines CoC1/DDP were transfected with miRNA-15a and miRNA-16 mimics and treated with cisplatin.qRT-PCR was used to detect the expression levels of miRNA-15a and miRNA-16 in normal CoC1/DDP cell group,cisplatin treated group,negative control group,miRNA-15a transfected group,miRNA-16 transfected group and overexpressed Bmi-1 plasmid.Western blot was used to detect the expression level of Bmi-1 in each group,CCK-8 and Annexin V/PI were used to detect cell survival and apoptosis,and γ-H2AX immunofluorescence was used to detect cell apoptosis.Results The CoC1/DDP ovarian cancer cell line shows low expression of miRNA-15a and miRNA-16,and high expression of Bmi-1 protein,which makes it resistant to cisplatin.When the levels of miRNA-15a and miRNA-16 are overexpressed,the Bmi-1 protein decreases(P<0.05),leading to a decrease in cell survival rate(P<0.05),a significant increase in DNA apoptosis(P<0.05),and more severe DNA damage(P<0.05).Overexpression of Bmi-1 plasmid can increase cell viability(P<0.05)and reduce the rate of cell apoptosis(P<0.05).Conclusion The Bmi-1 protein may be a target for the regulation of miRNA-15a and miRNA-16,and overexpression of miRNA-15a and miRNA-16 can increase the sensitivity of ovarian cancer cells to cisplatin by reducing the Bmi-1 protein.This provides a new idea for predicting molecular markers of cisplatin resistance in ovarian cancer and overcoming drug resistance targets in ovarian cancer.
ABSTRACT
Objective To investigate the relationship of Maternal-fetal interface local cytokine GM-CSF and its receptor with the occurrence of early spontaneous abortion. Methods From August 2009 to December 2011,we collected 30 villi tissue samples with artificial abortion and 30 villi tissue samples with spontaneous abortion. At the same time, we collected 30 villi tissue samples with artificial abortion and 30 decidua tissue samples with spontaneous abortion,and 30 decidua tissue samples with spontaneous abortion. The human chorionic gonadotropin ( HCG) was detected by radioimmunoassay in every group. The expressions of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR ) were detected by Immunohistochemical staining and Western Blot in the villi tissue and decidua tissue in every group. Results The concentration of HCG in the spontaneous abortion group was lower than that in the artificial abortion group ( <0.05) . The protein expressions of GM-CSF and GM-CSFR were found in villus and deciduas tissues in both groups. The protein expression levels of GM-CSFR in the villus tissues were higher in spontaneous abortion group than those in artificial abortion group ( <0.05), the protein expression of GM-CSF was upregulated, but there was no statistically significant difference between two groups. In deciduas tissues, the protein expressions of GM-CSF and GM-CSFR were upregualted in spontaneous abortion group ( <0.05) . Conclusions The suitable concentrations of GM-CSF and GM-CSFR in decidua tissue maintain the pregnancy continued. However,the higher concentrations of GM-CSF and GM-CSFR in the decidua tissue may be one of reasons of spontaneous abortion.