Clinical Application And Significance Of Human Mammaglobin In Diagnosis, Prognosis And Targeted Therapy Of Breast Cancer

BackgroundBreast cancer (BCa) is one of the most common female malignancies worldwide and with a heavy mortality. The morbility of breast cancer is prone to be leading of female malignancies in China. The micrometastasis induce local relapse and distant metastasis in breast cancer, therefore identification of micrometastasis is of great importance in clinical management and with significant predictive and prognostic value. Based on these concept, breast cancer should be considered as systemic disease and some molecular biomarkers should be needed in precision medicine for breast cancer. Human mammaglobin (hMAM) is a member of the secretoglobin superfamily and could be served as a novel molecular biomarker for diagnosis and prognostic evaluation of breast cancer. The discussion of hMAM expression and its significance will be of great clinical value in breast cancer.AbstractObjective:To detect expression pattern of hMAM in breast cancer and investigate its correlation with clinicopathological parameters of breast cancer patients. Furthermore we evaluate the potential possibility of hMAM as a novel molecular biomarker for diagnosis and prognostic assessment of breast cancer.Methods:The clinicopathological information of 172 patients admitted to Anhui Provincial Hospital for surgical treatment of breast neoplasm between Jan 2012 and Dec 2014 were retrospectively summarized, including primary breast invasive carcinoma, carcinoma in situ and benign neoplasm. All patients were received no any treatment before surgery. Immunohisto-chemical staining were conducted to detect expression pattern of hMAM and GCDFP-15. Then evaluate the correlation between expression of hMAM and expression of GCDFP-15 and clinicopathological characteristics in breast cancer. The expressions of hMAM in carcinoma in situ of breast, breast benign neoplasms, non-breast origin malignancies and normal tissues were also detected by immunohistochemistry respectively.Results:There are two expression patterns of hMAM in breast cancer, including located in cytoplasm and cell membrane. The positive ratio of hMAM expression in breast benign neoplasm, carcinoma in situ and invasive carcinoma of breast was 3.77%(2/53),20.51%(8/39) and 72.50%(58/80) respectively, which was significantly different (P<0.01). By contrast, there was no expression of hMAM in non-breast origin human malignancies and normal tissues. Furthermore, our results indicated that there was no association for the expression of hMAM with tumor grading (P=0.33). However, there was a significant correlation between the expression of hMAM and tumor clinical stage, axillary lymph node status and expression of GCDFP-15.Conclusions:hMAM protein could specifically detected in breast cancer and with highly expression level. hMAM could be considered as a novel molecular biomarker for diagnosis and prognosis of breast cancer. There might be cell membrane associated hMAM and it has potential providing a novel marker for targeted therapy of breast cancer.BackgroundThe highly mortality of breast cancer is attributed to its high capacity to micrometastasis via various approaches. Therefore identification and monitoring of micrometastasis is of great importance in clinical management and prognostic evaluation of breast cancer. However, there now is no specific and sensitive methodology for detecting micrometastasis. The incidence of so-called "Circulating Tumor Cells" (CTC) and the corresponding detection were considered as an impressive approaches for early diagnosis, clinical management and relapse monitoring of human solid malignancies, including breast cancer. Therefore several researches focused on the clinical application of CTC in breast cancer. There are three important factors associated with precision detection of CTC, that is isolation of CTC, detective approaches and molecular biomarker. Molecular biomarkers with tumor specificity, tumor sensitivity and organ specificity are essential for detection of CTC. hMAM is becoming a novel biomarker for breast cancer because of its characteristics. There are several methods used for isolation of solid tumors, but these approaches are all sophisticated and could not be widely applied. Peripheral blood mononuclear cells (PBMCs) were appropriate surrogate of CTC for enhancing clinical application of CTC in solid malignancies. There are growing body of evidence suggested that PBMC detection by RT-PCR in combination with specific molecular biomarker will be a promising approach for CTC detection of breast cancer, furthermore for precision diagnosis and prognostic evaluation of breast cancer.Recent researches demonstrated that hMAM mRNA highly expressed in peripheral blood of breast cancer patients and it has clinical significances in precision diagnosis of breast cancer. hMAM mRNA expression in peripheral blood is equal to discovery of CTC. Therefore detection of hMAM mRNA in peripheral blood has shown impressive value in dynamic evaluation of metastasis and predictive assessment in breast cancer.AbstractObjective:To establish methodology for detection of CTC in peripheral blood of breast cancer patients used hMAM as molecular biomarker, and evaluate the correlation of hMAM mRNA expression in peripheral blood with clinicopathological characteristics of breast cancer.Methods:The clinicopathological information of 172 patients admitted to Anhui Provincial Hospital for surgical resection of breast neoplasm between Jan 2012 and Dec 2014 were retrospectively reviewed, including primary breast invasive carcinoma, carcinoma in situ and benign neoplasm.75 breast cancer patients have no distant metastasis via radiological examinations when enrolling. All blood samples were collected before the initiation of any treatment. PBMCs were isolated from the blood samples and total RNA was extracted. SYBR Green RT-PCR were used to detect hMAM mRNA expression in combination with electrophoresis analysis. The expression profiles of hMAM mRNA in different breast neoplasms were analyzed and the association between the expression of hMAM mRNA and clinicopathological characteristics was evaluated.Results:The RT-PCR product length of hMAM and GAPDH was 105 bp and 154 bp respectively. Results were considered as positive when the target band (hMAM) was lighter than GAPDH band. The positive ratio of hMAM mRNA expression in breast benign neoplasm, carcinoma in situ and invasive carcinoma of breast was 0%(0/53), 51.3%(20/39) and 75.0%(60/80) respectively, which has significantly different between breast benign neoplasm and carcinoma in situ, additionally between breast benign neoplasm and breast invasive carcinoma (P<0.01). However, there was no discrepancy between carcinoma in situ and breast invasive carcinoma(P=0.10). The expression ratio of hMAM mRNA in peripheral blood of breast cancer patients with histological grade ?, grade? and grade III was 75.00%,75.76%and 73.91% respectively. The expression ratio of hMAM mRNA in peripheral blood of breast cancer patients with clinical stage ?,? and ? was 66.67%,68.00% and 95.24% respectively. These was no correlation between the expression of hMAM mRNA and tumor grade and clinical stage of breast cancer. By contrast, the expression ratio of hMAM mRNA in peripheral blood of breast cancer patients with axillary lymph node metastasis and without metastasis was 95.00% and 55.00% respectively. There has significant association for the expression of hMAM mRNA in peripheral blood with axillary lymph node positive status (P<0.01).Conclusion:hMAM mRNA could be specifically detected in peripheral blood with highly expression level in breast cancer patients. There was significant correlation between the expression of hMAM mRNA and axillary lymph node metastasis. Our data indicated that evaluation of PBMCs by using hMAM mRNA as molecular biomarker and using SYBR Green RT-PCR as detection approach could be effectively applied for metastasis monitoring and prognostic assessment of breast cancer.BackgroundBreast cancer is a complex and heterogeneous disease caused by multiple unknown factors, therefore there might be several molecular biomarkers associated with diagnosis and therapy of breast cancer. However, currently there are limited markers could be applied and some biomarkers of breast cancer used clinically still have shortcomings. That is the reason why precision diagnosis and targeted therapy could not efficiently guide the clinical management of breast cancer.hMAM is a breast cancer-specific molecular biomarker. Several evidences have shown the secretory nature of hMAM protein and there might have some of MAM proteins association with the membrane of breast cancer cells after secretion. Recent studies have provided robust evidences that hMAM protein could be expressed on membrane of breast cancer cells and membrane associated hMAM protein have potential important role in proliferation, invasion and metastasis of breast cancer. Therefore it is concluded that membrane-expressed hMAM protein might be a novel therapeutic target for precision management of breast cancer and will provide a new insight into the elucidation of molecular mechanism of breast cancer.AbstractObjectives:To confirm that hMAM protein could be expressed on membrane of breast cancer cells and provide preliminary evaluation of the effect of membrane associated hMAM on proliferation of ZR75-1 cells in vitro.Methods:Immunohistochemical staining were used to detect the expression of membrane associated hMAM in breast cancer in vivo and Western blot were conducted in vitro. MTT assay was used to evaluate the effect of anti-hMAM antibody on proliferation of ZR75-1 cells, meanwhile irrelevant antibody control, negative control and blank control were detected.Results:Membrane associated hMAM protein could be specifically recognized by 31A5 antibody. Results of immunohistochemistry were negative when non-breast origin malignancies and several human normal tissues were detected by 31A5 antibody. Western blot has shown that membrane associated hMAM moderately expressed in MDA-MB-231 cells and weakly expressed in ZR75-1 and MDA-MB415 cells. MTT assay demonstrated that the inhibition of ZR75-1 cell proliferation was induced by treatment of 31A5 antibody compared with control group (p<0.05) However, the inhibitory effect was not observed in other breast cancer cell lines and other time point.Conclusions:Our results demonstrated that there are some of hMAM proteins expressed on the membrane of breast cancer cells in vivo and in vitro and it has different expression level of membrane associated hMAM protein for various breast cancer cells. Membrane-expressed hMAM protein and its associated downstream signaling pathways might have enhanced role in proliferation of ZR75-1 cell. Some antagonist antibody against membrane associated hMAM have potential to block this pathway and inhibit ZR75-1 cell proliferation.