The Molecular Mechanism Of LncRNA CERS6-AS1/miR-125a-5p/BAP1 Axis Regulating Breast Cancer Cell Proliferation

Objective:This experiment is to explore the influence of lncRNA CERS6-AS1/miR-125a-5p/BAP1 axis on the occurrence and development of breast cancer,and to further explore its possible molecular mechanism.Methods:In the experiment,we will use bioinformatics analysis methods,RNA pull-down technology,fluorescence in situ hybridization(FISH)and dual luciferase reporter gene experiments to predict and confirm the interaction of lncRNA with miR-125a-5p.qRT-PCR was used to detect the expression of lncRNA,miR-125a-5p and BAP1 in normal breast tissues,tumor tissues and breast cancer cells,and nude mouse tumor-bearing experiments were used to verify the role of lncRNA in regulating breast cancer growth in vivo.Cell function experiments(EDU,flow cytometry apoptosis)were used to detect the effects of lncRNA and miR-125a-5p on the proliferation and apoptosis of breast cancer cells;Western Blotting was used to explore the expression of BAP1 protein in cells.Result: Through bioinformatics analysis,it is found that lncRNA CERS6-AS1 and miR-125a-5p have binding sites;the results of dual luciferase reporter gene experiment and RNA pull-down experiment show that lncRNA CERS6-AS1 and miR-125a-5p can bind;Fluorescence in situ hybridization experiments found that lncRNA CERS6-AS1 mainly exists in the cytoplasm of breast cancer cells.By using qRT-PCR technology,we found that in 17 pairs of matched breast cancer tissues and their adjacent tissues,lncRNA CERS6-AS1 was significantly highly expressed in tumor tissues and negatively correlated with miR-125a-5p.In vitro experiments found that when lncRNA CERS6-AS1 was knocked down,the proliferation ability of MDA-MB-231 and MDA-MB-468 cells was significantly decreased,and the proportion of apoptosis was significantly increased;and when lncRNA CERS6-AS1 was overexpressed,the mammary glands The proliferation ability of cancer cells is enhanced,and the proportion of apoptotic cells is reduced.In vivo experiments confirmed that the growth of transplanted tumors increased rapidly,and the volume and weight increased significantly after overexpression of lncRNA CERS6-AS1;the immunohistochemical results of transplanted tumors showed increased expression of Ki-67 and BAP1.Functional recovery experiments showed that the restoration of miR-125a-5p levels inhibited the proliferation promotion of CERS6-AS1 and the inhibition of cell apoptosis.Conclusion: CERS6-AS1 promotes the proliferation of breast cancer cells in vitro,inhibits apoptosis,and promotes the growth of transplanted tumors in nude mice in vivo.And as a ceRNA to regulate miR-125a-5p to regulate BAP1 to promote breast cancer cell proliferation.