Angiogenesis And Lymphangiogenesis In Human Breast Cancer Metastasis And Prognostic Significance

Distant metastasis is the leading cause of cancer death and responsible for the poor therapeutic reactions and miserable prognosis in breast cancer patients. Hematogenous metastasis and lymphatic system dissemination are the most important metastasis pathways and vascular endothelial growth factor (VEGF) is the most effective mitogen for the vascular endothelial cells and critical factor on breast cancer angiogenesis and metastasis. Several novel members of the VEGF family as VEGF-C and VEGF-D and their receptor flt-4 (VEGFR-3)are not only important regulators of lymphangiogenesis in vivo but also enhance lymphatic metastasis associated with prognosis, direct evidence that breast cancers are able to activate tumor lymphangiogenesis, a phenomena that long neglected in the past has been demonstrated while the underlying mechanism are poorly understood. Chemokines are a family of short peptides that regulate angiogenesis, promote cancer cells chemoattractic migration and metastasis and has specific expression spectrum in some breast cancer cell lines and patients also been reported.Human inflammatory breast cancer (IBC) is a highly aggressive form of locally advanced breast cancer with miserable prognosis due to its propensity to disseminate via the dermal lymphvascular invasion exhibiting skin erytherma and distant metastasis at early stage, human IBC cell line may provide an excellent cell model for further investigation of the molecular machinery responsible for this extremely malignancy and understanding of metastasis for better management.Subjectives To establish the human inflammatory breast cancer cell line, as an excellent cell model for further investigation of this extremely malignancy and understanding of metastasis; investigate the expression of VEGF, VEGF-C and flt-4 in breast cancer and its relationship with lymphatic metastasis and explore the significance of lymphangiogenesis on metastasis via lymphatics also. We study the expression of chemokines and VEGFs family members in breast cancer patients andanalyze the correlations with tumor angiogenesis and prognosis. Methods Development and selection for biological stable human inflammatory breast cancer cell line by in vitro series culture method and clarify its identical characteristics.Paraffin-embedded breast cancer specimens undergone radical mastectomy with lymph node dissection were investigated by immunohistochemical staining method for VEGF,VEGF-C,flt-4 expression and analyze their relationship with lymphatic metastasis.Multiple-probe RNase Protection assay (RPA) method detect the expression of chemokines and VEGFs family in breast cancer patients and explore its correlations with clinical pathological significance. Enzyme linked immunosorbent assay (ELISA) kits were used with 93 breast cancer patients and 20 health controls, and their clinical pathological significance analysis.Results 1.Human stable IBC cell line were established nominated as IBC-1, display polygonal cell shape and aggregated population in culture, chromosome analysis revealed sub-triploid karyotype, the number of chromosomes per cell varied from 49~61(71.6%) ,western blot studies show E-cadherin a high expression. the subcutaneously inoculated tumorigenicity was 100% and spontaneous lung metastasis in nude mice within 4 weeks ,The flow cytometry shows similar cell cycle distributions both in IBC-1 and xenograft tumor cells .2. Positive expression rate of VEGF-C(38/82, 46.3%)and flt-4(41/82, 50%) in lymph node-positive group were significant higher than node-negative group(P﹤0.05). Furthermore, the correlation was found between the expression of VEGF-C and flt-4 also(P﹤0.01).3.The flt-1, flt-4 mRNA levels in breast cancers were significant higher than paracancerous tissues control; chemokines as IL-8, IP-10, MIP-1α, MIP-1β were significant higher in tumor than paracancerous pecimens(P﹤0.05). Significant correlations were also founded between VEGF, Ang-1 mRNA expression and CD31, CD105. The novel microvessel density mar...