BACKGROUND: The early stages of tumor bone metastasis are closely associated with changes in the vascular niche of the bone microenvironment, and abnormal angiogenesis accelerates tumor metastasis and progression. However, the effects of lung adenocarcinoma (LUAD) cells reprogrammed by the bone microenvironment on the vascular niche within the bone microenvironment and the underlying mechanisms remain unclear. This study investigates the effects and mechanisms of LUAD cells reprogrammed by the bone microenvironment on endothelial cells and angiogenesis, providing insights into the influence of tumor cells on the vascular niche within the bone microenvironment. METHODS: The culture media from bone-metastatic LUAD cell A549-GFP-LUC-BM3 (BM3-CM) and A549-GFP-LUC (A549-CM) were separately applied to human umbilical vein endothelial cell (HUVEC). A colony formation assay, scratch assay, and tube formation assay were conducted to evaluate the proliferation, migration, and angiogenesis of HUVEC. Gene set enrichment analysis (GSEA) was conducted to identify enriched pathways, while reverse transcription quantitative polymerase chain reaction (RT-qPCR) and enzyme linked immunosorbent assay (ELISA) were performed to quantify hepatocyte growth factor (HGF), a protein that plays a crucial role in angiogenesis. Furthermore, the pivotal function of HGF and its underlying molecular mechanisms have been substantiated through the utilisation of recombinant proteins, neutralising antibodies, pathway inhibitors, immunofluorescence staining, and Western blot. RESULTS: BM3-CM demonstrated a more pronounced impact on the proliferation, migration, and angiogenesis of HUVEC compared to A549-CM. Bioinformatics analysis, combined with in vitro experiment, demonstrated that the secretory protein HGF was significantly elevated in BM3 cells and BM3-CM (P<0.05). The addition of HGF neutralizing antibodies to BM3-CM inhibited the promoting effect of BM3-CM on HUVEC (P<0.05), while the addition of recombinant HGF to A549-CM reproduced that promoting effect of BM3-CM on HUVEC (P<0.05). HGF can enhance the activation of YAP (Yes-associated protein) in HUVEC, and this promotion effect may be achieved by activating Src and activating YAP into the nucleus (P<0.05), but this effect can be inhibited by HGF neutralizing antibodies (P<0.05). Furthermore, the addition of recombinant HGF to A549-CM can recapitulate the YAP activation effect of BM3-CM in HUVEC (P<0.05). CONCLUSIONS Bone microenvironment reprogrammed bone-metastatic LUAD cells BM3 promote the proliferation, migration, and angiogenesis of HUVEC through the HGF/YAP axis, potentially playing a significant role in the modifications of the vascular niche.
Humans ; Hepatocyte Growth Factor/genetics* ; Signal Transduction ; Neovascularization, Pathologic/genetics* ; Human Umbilical Vein Endothelial Cells ; Bone Neoplasms/blood supply* ; Adenocarcinoma of Lung/genetics* ; Adaptor Proteins, Signal Transducing/genetics* ; Lung Neoplasms/genetics* ; Cell Movement ; Cell Proliferation ; YAP-Signaling Proteins ; Transcription Factors/genetics* ; Cell Line, Tumor ; Tumor Microenvironment ; Angiogenesis

Osseous hemangioma is a benign vascular tumor, and it usually occurs in the vertebrae and the skull. However, hemangiomas of flat bones are rare, and there are very few reports that describe the radiologic findings of osseous hemangioma of the ilium. We report a unique case of large cavernous hemangioma mimicking a chondrogenic malignant bone tumor originated from the ilium in a 22-year-old female. The mass showed stippled calcifications, heterogeneous enhancement with thick septa and enhanced soft tissue components on CT and MR, and also this mass demonstrated heterogeneous 2-fluoro [fluorine-18]-2-deoxy-D-glucose (18F-FDG) uptake on 18F-FDG PET/CT.
Bone Neoplasms/radionuclide imaging ; Diagnosis, Differential ; Female ; Fluorodeoxyglucose F18/*diagnostic use ; Hemangioma, Cavernous/*radionuclide imaging ; Humans ; Ilium/*blood supply ; Magnetic Resonance Imaging ; Positron-Emission Tomography and Computed Tomography ; Radiopharmaceuticals/*diagnostic use ; Young Adult

OBJECTIVETo study the effects of brucine on vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in a nude mouse model of bone metastasis due to breast cancer, and to assess the possible antitumor mechanism of brucine.

METHODSA syringe needle was used to directly inject 0.2 mL monoplast suspension (with 2×10(5) human breast cancer cells contained) into the bony femoral cortex of the right hind leg for modeling. Twenty-five nude mice were randomized into five groups and administered with an intraperitoneal injection of saline or drug for 8 consecutive days: model group (0.2 mL normal saline), low-dose brucine group (1.73 mg·kg(-1)), medium-dose brucine group (3.45 mg·kg(-1)), high-dose brucine group (6.90 mg·kg(-1)), and thalidomide group (200 mg·kg(-1)). Diet and activity were recorded, and the tumors were harvested 5 weeks later. The percentage of VEGF-positive cells was determined with hematoxylin and eosin staining and immunohistochemical staining, and MVD expression was determined by optical microscopy.

RESULTSThe VEGF expressions in brucine- or thalidomide-treated mice were significantly reduced as compared with mice in the model group (P <0.01). There were no significant difference between the high-dose brucine group and the thalidomide group (P >0.05). Significant difference was between the high- and low-dose brucine group P<0.05). Further, VEGF expression was significantly increased in the low- and medium-dose brucine groups compared with the thalidomide group (P <0.05). The MVD values in the three brucine and thalidomide groups were significantly lower than that in the model group (P <0.01). The MVD values in the medium- and high-dose brucine groups were not significantly different from those in the thalidomide group (P >0.05), while the MVD value showed a significant increase in the low-dose group compared with the thalidomide group (P <0.05).

CONCLUSIONBrucine could inhibit the growth of breast cancer to bone metastases, possibly by inhibiting tumor angiogenesis.

Animals ; Bone Neoplasms ; blood supply ; metabolism ; secondary ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; drug effects ; pathology ; Strychnine ; analogs & derivatives ; pharmacology ; therapeutic use ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo study the methods and effects of the reconstruction of tumor-induced bone defects with vascularized fibula graft.

METHODSFrom Oct. 1996 to Jan. 2005, 89 patients with bone defects were treated with fibula graft using different methods. Among the patients, 48 patients were male and 41 patients were female, ranging in age from 12 to 67 years, with an average of 35 years. Thirty-five patients were treated with inlay bone grafting after excision of focus, 15 patients were treated with single or double-strut fibula graft after tumor resection, 16 patients were treated with fibular head graft for juxta articular tumor resection, and 23 patients were treated with double-strut fibula combined with iliac graft after marginally resection.

RESULTSEnneking evaluation system was used to evaluate therapeutic effects. Among 35 patients treated with inlay bone graft after excision, 29 patients were followed up, and 26 patients got an excellent result, 1 good and 2 poor. Among 15 patients treated with single or double-strut fibula graft after tumor resection, 12 patients were followed up, and 8 patients got an excellent result, 1 good, 1 poor and 2 bad. Among 16 patients treated with fibular head graft for juxta articular tumor resection, 12 patients were followed up, and 7 patients got an excellent result, 3 good,1 poor and 1 bad. Among 23 patients treated with double-strut fibula combined with iliac graft after marginally resection, 17 patients were followed up, and 11 got an excellent result, 3 good, 1 fair and 2 bad.

CONCLUSIONThese treatment methods can greatly enrich the treatment methods for bone tumor, and satisfy the reconstruction after bone tumor excision in different position of the four limbs. These methods are reliable and can be chosen according to disease types.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; surgery ; Bone Transplantation ; methods ; Child ; Female ; Fibula ; blood supply ; transplantation ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Treatment Outcome ; Young Adult

PURPOSEWe have previously shown that osteosarcomas have states of increased interstitial fluid pressure (IFP) which correlate with increased proliferation and chemosensitivity. In this study, we hypothesized that constitutively raised IFP in osteosarcomas regulates angiogenesis.

MATERIALS AND METHODSSixteen patients with the clinical diagnosis of osteosarcomas underwent blood fl ow and IFP readings by the wick-in-needle method at the time and location of open biopsy. Vascularity was determined by capillary density in the biopsy specimens. We performed digital image analysis of immunohistochemical staining for CD31, VEGF-A, VEGF-C and TPA on paraffin-embedded tissue blocks of the biopsy samples. Clinical results were validated in a pressurised cell culture system.

RESULTSIFPs in the tumours (mean 33.5 +/- SD 17.2 mmHg) were significantly higher (P = 0.00001) than that in normal tissue (2.9 +/- 5.7 mmHg). Pressure readings were significantly higher in low vascularity tumours compared to high vascularity tumours (P <0.001). In the osteosarcoma cell lines, growth in a pressurised environment was associated with VEGF-A downregulation, VEGF-C upregulation and TPA upregulation. The reverse was seen in the OB cell lines. Growth in the HUVEC cell line was not significantly inhibited in a pressurised environment. Immunohistochemical assessment for VEGF-A (P = 0.01), VEGF-C (P = 0.008) and TPA (P = 0.0001) translation were consistent with the findings on PCR.

CONCLUSIONOur data suggest that some molecules in angiogenesis are regulated by changes in IFP.

Adolescent ; Angiogenic Proteins ; physiology ; Bone Neoplasms ; blood supply ; Cells, Cultured ; Extracellular Fluid ; physiology ; Female ; Humans ; Male ; Neovascularization, Pathologic ; Osteosarcoma ; blood supply ; Pressure

OBJECTIVETo analyze the relationship between Duffy antigen receptor for chemokines (DARC) and the metastasis potential in human breast cancer. METHODS Breast cancer tissue sections from 75 patients, grouped according to the local lymph node status were examined immunohistochemically for protein level of DARC. Microvessel density (MVD) was counted by endothelial cells immunostained using anti-CD34 antibody.

RESULTSStrong positive DARC immunostaining in lymph node negative and positive groups was detected in 31 cases (81.6%) and 18 cases (48.6%), respectively (P < 0.01). MVD was (35.67 +/- 17.96)/HP and (53.38 +/- 20.29)/HP in DARC strong positive and less positive cases (P < 0.01). In those patients with lung, bone, hepatic distant metastasis (13 cases), 9 cases (69.2%) were DARC less positive, 4 cases (30.8%) were DARC strong positive. The correlation coefficient was -0.412 between DARC expression and MVD and the corresponding value was -0.346 between DARC expression and lymph node status and -0.333 between DARC expression and distant metastasis in breast cancer.

CONCLUSIONDARC may play a negative role in the process of neoangiogenesis, and probably has an association with the lymph node status.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; analysis ; Bone Neoplasms ; metabolism ; secondary ; Breast Neoplasms ; blood supply ; metabolism ; pathology ; Down-Regulation ; Duffy Blood-Group System ; metabolism ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; secondary ; Lung Neoplasms ; metabolism ; secondary ; Lymphatic Metastasis ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism ; Survival Analysis

OBJECTIVETo investigate the effect of antisense vascular endothelial growth factor (VEGF)(121) cDNA transfection on the growth of K562 cells in nude mice.

METHODSK562 cells transfected with the antisense (AS) or sense (S) VEGF(121) cDNA, and the vector (V, pcDNA3) alone were transplanted subcutaneously into nude mice and the growth of the transfected cells in vivo was investigated. The effects of transfected K562 cells on human bone marrow endothelial cells (BMEC) were analyzed by MTT assay, the microvessel density (MVD) in tumor mass by vWF immunohistochemistry stain.

RESULTSK562/V tumor grew more slowly [(207.5 +/- 192.9) mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.05] and K562/S tumor more rapidly than K562/V tumor did [(1 174.6 +/- 508.7)/mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.01]. K562/S cell culture supernatant was more strongly in promoting the proliferation of BMEC than K562/V supernatant did, but K562/AS supernatant resulted in a marked decrease of the promoting effect as compared with K562/V's. The MVDs in K562/AS, K562/S, and K562/V tumors were [(11.0 +/- 7.6)/0.72 mm(2) vs (50.8 +/- 11.7)/0.72 mm(2) vs (18.9 +/- 7.0)/0.72 mm(2)], respectively.

CONCLUSIONSAntisense VEGF(121) cDNA transfected K562 cells show growth retardation in transplanted nude mice, decrease of tumor MVD, and decrease of promoting BMEC proliferation capacity.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Division ; genetics ; physiology ; Culture Media, Conditioned ; pharmacology ; DNA, Antisense ; genetics ; DNA, Complementary ; genetics ; Endothelial Growth Factors ; genetics ; physiology ; Endothelium, Vascular ; cytology ; drug effects ; Female ; Humans ; K562 Cells ; Lymphokines ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; blood supply ; genetics ; pathology ; Neovascularization, Pathologic ; genetics ; physiopathology ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors