Study Of Correlation Between The Angiogenesis Regulatory Related Proteins And Breast Cancer

Breast cancer is one of common cancers which seriously affect women’s health. In China, the incidence of breast cancer increased year by year, above all in some areas the incidence ranked the first of all the females malignant tumors, and the age of onset tends to be younger. Breast cancer is multi-vessel, similar to the majority of tumors, its occurrence and development as a result of multiple genes, multi-stage process involves a variety of factors including of the external environment and genetics. Etiology and prognostic factors are not entirely clear. Tumor angiogenesis is an important factor leading to tumor recurrence and metastasis. Folkman was first proposed in1970that tumor growth depends on angiogenesis. Based on the theory, the broken balance between angiogenesis factor and angiogenesis inhibitory factor leads to tumor angiogenesis of breast cancer. From a biological point of view, angiogenesis regulatory proteins have been identified as an important regulator of tumor growth for breast cancer. Angiogenesis regulatory proteins, which are implicated in tumor growth and invasion, represent desirable therapeutic and diagnostic targets. Platelets play a major role in hemostasis, as well as in tissue repair, maintenance of endothelium, and vascular tone, tumors are wounds that do not heal, the contribution of platelets to the progression of cancer is an emerging area of research interest. Numerous angiogenesis regulatory proteins are present in platelets. Under physiological conditions, angiogenesis regulatory proteins are sealed in circulating platelets, which are relatively stable. When the tumor occurs, the storage proteins are released, resulting in a significant change. Studies have found elevated levels of vascular growth factor in the patient’s plasma and tumor tissue, however, of interest is the observation that in some cancers, many angiogenic factors in tumor-bearing animals’ platelet concentration is much higher than in plasma, serum and non-malignant tumors. Therefore, regulating platelet protein in platelet may be a more sensitive indicator than in plasma and serum for responding tumor occurrence and development.This study aimed to investigate the expression of angiogenesis regulatory proteins in platelets and tumor tissue of breast cancer patients and its correlation with clinicopathological features, attempted to find possible impact to early diagnosis and prognosis of breast cancer, and thus a more accurate assessment of prognosis and better guide clinical treatment. This study is divided into two parts.Part1Objective:Platelets play an important role in tumor angiogenesis and growth and are the main transporters of several angiogenesis regulators. Here, we analyzed the levels of angiogenic factors sequestered by circulating platelets in breast cancer patients and age-matched healthy controls.Methods:Platelet pellets and platelet-poor plasma were collected by routine protocols. Vascular endothelial growth factor (VEGF), platelet-derived growth factor BB (PDGF-BB), thrombospondin-1(TSP-1), platelet factor4(PF4), and transforming growth factor-β1(TGF-β1) were measured by enzyme-linked immunosorbent assays, and protein concentrations were normalized to platelet numbers by an actin enzyme-linked immunosorbent assay.Results:Angiogenesis-associated expression of VEGF (2.1pg/106vs.0.9pg/106platelets,p<0.001), PF4(21.2ng/106vs.10.2ng/106platelets,p<0.001), PDGF-BB (42.9pg/106vs.19.1pg/106platelets,p<0.001), and TGF-β1(15.3ng/106vs.4.3ng/106platelets,p<0.001) differed between platelet pellet samples from cancer and control subjects. The areas under the receiver-operating characteristic curves were>0.7. VEGF, PDGF-BB, and TGF-β1levels were higher in platelets in stage Ⅲ+Ⅳ, lymph node1-3, human epidermal growth factor receptor-2(+), and basal-like subtypes, which indicate poor prognosis (p<0.05).Conclusions:PF4, VEGF, PDGF-BB, and TGF-β1levels were elevated in platelet pellets versus platelet-poor plasma in breast cancer, suggesting that platelet levels are better indicators for disease diagnosis than plasma levels. VEGF, PDGF-BB, and TGF-β1levels in platelets may be associated with breast cancer prognosis.Part2Objective:Tumor angiogenesis is essential for the growth and spread of breast cancer cells. Therefore, the aim of this study was to investigate the expression of PDGF-BB, TGF-β1and VEGF in breast cancer, to analyze their clinical significance and the correlation with the levels of the three proteins in PPP, and to explore these proteins possible roles in the process of occurrence, invasion and metastasis of breast cancer. Method:133cases paraffin-embedded tissue specimens during2011to2013were collected from the department of Pathology of Liaocheng People’s Hospital including80cases of invasive ductal carcinoma and53cases of adjacent tissues. All the cases were first surgery and no preoperative adjuvant therapy:chemotherapy and endocrine therapy. VEGF, PDGF-BB and TGF-β1of all samples were detected by SP immunohistochemical staining method. Microvessel density (MVD) marked CD34were detected by SP immunohistochemical staining.Results:In80cases of breast cancer tissues, the positive rates of VEGF, PDGF-BB and TGF-β1protein were76%,68%and81.9%respectively, significantly higher than the adjacent normal tissues (P<0.05). The level of VEGF protein were positively correlated with clinical stage of breast cancer, lymph node status, positive expression of ER, MVD, etc.. TGF-β1protein expression was significantly increased in the following state:late clinical stage, lymph node metastasis, MVD higher, with statistical significance (P<0.05). PDGF-BB were significantly correlated with lymph node status, MVD, etc.(P<0.05). In80cases of breast cancer patients, expression of PDGF-BB, TGF-β1and VEGF was significantly higher in smoking group than that of in non-smoking group. The expression rates of PDGF-BB, TGF-β1and VEGF were45%、56.2%、52.5%respectively, and the difference was statistically significant (P <0.05). Expression of VEGF、PDGF-BB and TGF-β1protein in breast cancer tissue were moderate positive correlation with that of in platelet granule, Correlation coefficients were0.482,0.599,0.458respectively(P<0.05).Conclusion:Angiogenesis-related factor (VEGF, PDGF-BB and TGF-β1) expressed at high levels in the tissues of breast cancer, and positively correlated with corresponding protein in platelet granule. These three proteins might play an important role in the process of tumor growth, invasion and metastasis, and correlate with prognosis. Smoking can improve the expression of tumor angiogenesis regulatory proteins, and thereby promote neovascularization of tumor. Smoking may be involved in the occurrence and metastasis of breast cancer. Combination of angiogenesis markers both in tumor tissues and platelet granule can help screening high-risk breast cancer patients clinically.