Semaphorin 4C Mediates Tumor Lymphatic Metastasis And Serves As A Biomarker For Breast Cancer

Objective: Tumor metastasis is the leading cause of death in patients with cancer,we aimed to investigate whether and how semaphorin4C(SEMA4C)may play a role in tumor metastasis.And we aimed to evaluate whether serum SEMA4 C can early diagnose breast cancer.Methods: The c DNA microarray analysis following Lymphatic microdissection determined the differences in lymphatic endothelial cells.Differences in m RNA and protein levels were measured by real-time quantitative PCR and Western blotting(WB).Enzyme-linked immunosorbent assay(ELISA)was used to examine blood samples and cell culture supernatants.Immunofluorescence and immunohistochemistry were used to assess the localization and expression of SEMA4 C.Lymph node metastases were imaged in vivo.The effect of SEMA4 C in vitro on lymphangiogenesis was observed by migration and tube formation experiments of lymphatic endothelial cells.We conducted a large sample of retrospective studies.Between September 2014 and June 2016,we collected 948 breast benign tumors at the Department of breast surgery and physical examination center of Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology.Serum samples from patients and healthy women are collected next to the patient's medical record information or called for counseling,and serum SEMA4 C values are measured at the same time.Finally,the accuracy of the diagnosis is calculated using the receiver operating curve(ROC).Results: Tumor-associated lymphatic endothelial cells expressed high levels of SEMA4 C and produced secreted SEMA4 C,which had been demonstrated in both intracellular and culture supernatants.High secretory SEMA4 C was detected in the sera of breast and cervical cancer patients.It revealed that SEMA4 C mediated lymphangiogenesis in different ways from the migration and proliferation tumor cells.We detected serum SEMA4 C values in 948 women(449 pre-operative breast cancer specimens,182 post-operative breast cancer specimens,206 pre-operative benign breast tumor specimens,and 111 healthy control specimens),and performed clinical pathological analysis.Serum levels of SEMA4 C in patients with invasive breast cancer were found to be significantly higher than in all controls.After drawing the ROC curve,the SEMA4 C cut-off point for optimal diagnosis of breast cancer was 5.00 ng/m L.Comparison of breast cancer with healthy controls: AUC: 0.807,(95% CI: 0.76-0.86),sensitivity: 62.58%,specificity: 81.08%;breast cancer compared with benign breast disease: AUC: 0.748,(95% CI: 0.71-0.79),sensitivity was 62.36%,specificity was 71.36%;breast cancer compared with non-breast cancer: AUC:0.768(95% CI: 0.73-0.80),sensitivity and specificity were 62.58% and 74.76%,respectively.In addition,monitoring of serum SEMA4 C after treatment showed that the level of SEMA4 C decreased,but did not return to the level of a healthy control.Conclusions: SEMA4 C can promote tumor lymphatic metastasis,and serum SEMA4 C may act as a new protein biomarker for breast cancer and cervical cancer.Serum SEMA4 C has a potential to be a novel protein biomarker for early detection and diagnosis of breast cancer.