TLR4 Signaling In MiR-21 Expression And Proliferation/Apoptosis In Ovarian Cancer

Background:Epithelial ovarian cancer is one of the most common gynecological cancer in the global,the degree of deterioration of the disease is relatively fast,in the past year,about140,000 cases of all patients died.Many patients can not be founded in advanced stage for the no obvious symptoms in the early stage,and the lack of an effective method in the early diagnosis.After many related research survey found that: micro RNA(mi RNA)is produced with many kinds of disease and its further development are closely relevance.Numerous studies in the past few years among the relevant researchers have learned that in which the mi RNA is involved in many malignant tumors produce their development process,which is similar to the presence of functional oncogenes or tumor suppressor genes.It has been reported that some of mi RNA regulating abnormal cell proliferation,which also includes ovarian cancer.TLRs are a class of highly conserved pattern recognition receptors mediate the repair of the body.When TLRs signal is activated when the excessive activation or not enough time,it will lead to bodily functions appear abnormal performance,which would seriously have some severe disease.Inflammatory tumors often a key factor in their development of one of them, which is also the main features of tumor immune microenvironment,cancer itself is a process of abnormal body injury.TLR4 is a member of Toll-like receptor family of a class of innate immune molecules,it was highly expressed in a variety of malignant tumor cells,it has a very close relationship with apoptosis of tumor cells.Objective:This study was designed to observe the expression of mi R-21 in ovarian cancer tissues,and analyze the impact of mi R-21 expression changes on poptosis and proliferation of ovarian cancer cell.We explored the impact of signaling activation on expression and regulaition of mi R-21,and the effect on the biological behavior.Designed to understand the mechanism of inflammation-related cancer and provide a reliable basis for ovarian cancer diagnosis and targeted therapy.Methods:(1)We detected the expression of mi R-21 and TLR4 m RNA by q RT-PCR in ovarian cancer tissues and corresponding normal tissue;detected the expression of TLR4 protein by western blot in ovarian cancer tissues and corresponding normal tissue;analysis the correlation of mi R-21 and TLR4 m RNA between clinicopathological parameters.(2)mi R-21 mimics was transfected into SKOV-3 cells,in order to enhance the expression of mi R-21;we detected the expression of mi R-21 after transfection by q RT-PCR;detected the multiplication efficiency of SKOV-3 cells by MTT assay after transfecting mi R-21 minics;Annexin V-fitc/pi staining assay and flow cytometry was used to detect the poptosis in 0 h,24 h,48 h,72 h,96 h after transfection;detected the neoplasm invasion after transfection.(3)Ovarian cancer SKOV-3 cells were stimulated with lipopolysaccharide(LPS)after a preincubation and were simulated with NF-?B inhibitor PDTC following for 30 min.At 0 h,24 h,48 h,72 h,96 h after LPS stimulation,levels of mi R-21 were measured by q RT-PCR,phospho-NF-?Bp65 was determined by western blot analysis.Apoptosis,cell viability and invasion in SKOV-3 cells transfected with mock(control)or mi R-21 inhibitor were assessed by Annexin V / PI staining,MTT and Transwe II assay,respectively.Result:(1)The expression of mi R-21 in ovarian cancer was significantly higher than its expression in control group(P<0.01);the expression of TLR4 m RNA in ovarian cancer was significantly higher than its expression in control group(P<0.01);mi R-21 and TLR4 m RNA expression is closely related to histological grade and lymph node metastasis of ovarian cancer.(2)The espression of mi R-21 in SKOV-3 cells was significantly increased after transfecting mi R-21 mimic(P<0.01);mi R-21 can promote the proliferation of SKOV-3cells and inhibit apoptosis,and improving their invasiveness.(3)LPS induced significant increasement of mi R-21 level and NF-?B activation in SKOV-3 cells in a time-dependent manner(P<0.05),which were significantly attenuated by NF-?B inhibitor PDTC.Transfection of SKOV-3 cells with mi R-21 inhibitor resulted in a significant increase in apoptosis(P<0.05)and a significant decrease in cell growth and invasion,as compared with transfection with mock.Conclusion:(1)The increase of mi R-21 and TLR4 m RNA had a close relationship to histological grade and lymph node metastasis of ovarian cancer.(2)mi R-21 can promote proliferation and invasion,but inhibiting apoptosis in ovarian cancer SKOV-3 cells.(3)Activation of TLR4 signaling pathways may up-regulate mi R-21 through NF-?B activation in ovarian cancer SKOV-3 cells.Targeted inhibition of mi R-21 with a sequence-specific inhibitor can effectively induce apoptosis and suppress SKOV-3 cell growth and invasion,thus having a great potential in the treatment of ovarian cancer.