Preliminary Study On Expression And Function Of Fibulin-3 In Breast Cancer

Background and Purposes:Breast cancer(BC)is the most common cancer and leading cause of cancer death in female.Although systemic treatment for primary tumor have improved the prognosis of breast cancer patients significantly,it is less effective in treating metastatic breast cancer.Death from metastatic diseases in most patients with breast cancer is still the leading cause of death.Therefore,waging an in-depth research to explore the molecular mechanisms of metastasis of breast cancer,is of great importance in the diagnosis and treatment of metastatic breast cancer.Fibulin-3,belongs to the Fibulin glycoprotein family,is a widespread extracellular matrix protein.Fibulin-3 would affect the cell physiological functions such as metabolism,proliferation,and migration by regulating the interaction between cells and extracellular environment.Several studies have found that the expression level of Fibulin-3 is related to the occurrence and development of malignant tumors such as lung cancer,ovarian cancer,and liver cancer,but its relationship with breast cancer is rarely reported.This subject was to explore the relationship between Fibulin-3 and the prognosis of breast cancer patients by investigating the relationship between the expression of Fibulin-3 in breast cancer and the clinicopathological characteristics of patients.Additionally,we observed the effect of Fibulin-3 on the migration of breast cancer cells by interfering its expression.Methods:1.We detected differentially expressed genes in human Breast Cancer MDA-MB-231 Cell Line and MDA-MB-231 HM high metastatic cell Line by High-Throughput Sequencing.2.We investigated the expression of Fibulin-3 in 20 cases of breast cancer tissues and 20 cases of paired adjacent normal tissues by qPCR.3.We detected the expression of Fibuln-3 in 204 cases of breast cancer samples in tissue microarray by immunohistochemistry,and analyzed its relationship with clinicopathological features and prognosis of the breast cancer patients.4.QPCR and Western-Blot were used to detect the expression level of Fibulin-3 in human breast cancer cell line and human breast cancer cell line MCF-10 A.5.We constructed the Fiblin-3 knockdown cell lines and Fibulin-3 overexpressing cell lines,and verified that whether these cell lines have been successfully constructed by qPCR and Western-Blot.The relationship between Fibulin-3 and migration ability of breast cancer cells was verified by cell migration assay and cell scratch test.6.SPSS22.0 statistical software was used to perform the data analysis,X—±s was used to represent the measurement data.One-way-ANOVA,two-way-ANOVA,t-test and chi-square test were used to analyze the experimental data.Using α=0.05 as the test level,P<0.05 as the criterion of significant difference.Kaplan-Meier was used to analyze the prognosis of patients.Results:1.The expression Flibulin-3 in MDA-MB-231 HM high metastatic cell line was lower than that in MDA-MB-231 cell line(P<0.05).2.The expression of Fibulin-3 in cancer tissues was significantly lower than that in adjacent normal breast tissues(P<0.05).3.The expression of Fibulin-3 is significantly correlated with lymph node status in breast cancer patients(P=0.018).Kaplan-Meier survival analysis showed that patients with high Fibulin-3 expression had better prognosis.Cox regression analysis showed that Fibulin-3 is an independent prognostic marker for breast cancer.4.Compared with normal human breast cell line MCF-10 A,Fibulin-3 expression is down regulated in human breast cancer cell line.5.Cell migration experiment and cell scratch test confirmed that the migration ability of Fibulin-3 overexpressing breast cancer cell lines was decreased,while the migration ability of Fibulin-3 knockdown breast cancer cell lines was increased.Conclusion:1.Fibulin-3 is related to the occurrence and development of breast cancer.2.Fibulin-3 could be regarded as an independent predictor of prognosis in patients with triple negative breast cancer.3.Fibulin-3 could inhibit the migration of triple negative breast cancer cells...