OBJECTIVETo observe microvessel density(MVD), epithelial stromal vascular cuffing(VC) and expression of vascular endothelial growth factor(VEGF) in human cervical carcinomas and to clarify their significance in the invasion and metastasis of cervical carcinoma.

METHODSVEGF and CD34 were stained immunohistochemically (SP) in 57 cases of cervical carcinoma (30 cases of squamous cell carcinoma, 20 of adenocarcinoma 7 of glandular and squamous cell carcinoma), 29 cases of cervical intraepithelial neoplasia (CIN) and 16 cases of normal cervices, meanwhile, MVD and VC were also assayed.

RESULTSThere were significant differences among the above 5 groups for MVD P<0.01 . The VC pattern showed a significant difference between cervical carcinoma and CIN or control group P<0.01). The positive rates of VEGF in normal cervical epithelium, CIN, squamous cell carcinoma, adenocarcinoma, glandular and squamous cell carcinoma were 18.8% 3/16, 82.8% 24/29), 93.3% 28/30), 100% 20/20 and 7/7(100%), respectively. There were significant differences between these cervical lesion groups and the control group(P<0.001). The MVD showed significant differences between the positive pelvic node metastasis and negative pelvic node metastasis P<0.05). There was no significant correlation between the expression of VEGF and the tumor diameter, clinical stage, pathologic grade and pelvic node metastasis.

CONCLUSIONThe expression of VEGF may play an important role in the angiogenesis of cervical carcinoma. Degree of malignancy of cervical carcinoma has a close association with microvessel density.

Adult ; Aged ; Aged, 80 and over ; Endothelial Growth Factors ; analysis ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; analysis ; Lymphatic Metastasis ; Lymphokines ; analysis ; Microcirculation ; Middle Aged ; Uterine Cervical Neoplasms ; blood supply ; chemistry ; pathology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

To investigate the effect of arsenic trioxide (As(2)O(3)) on vascular endothelial growth factor (VEGF) expression in different courses of chronic myeloid leukemia (CML), VEGF level was measured with ELISA in the cultural supernatants of bone marrow mononuclear cells from CML patients. The results showed that supernatants of cultured bone marrow cells from 35 CML patients (20 chronic, 8 accelerated and 7 blast crisis phases) contained significantly higher VEGF levels (649.16 +/- 382.20 pg/ml, 560.27 +/- 409.14 pg/ml and 587.18 +/- 415.28 pg/ml, respectively) than that in 15 normal control samples (152.16 +/- 150.09 pg/ml; P < 0.01), but no significant differences were found in VEGF levels among different phases of CML. The bone marrow cells treated with As(2)O(3) (5 x 10(-6)mol/L) for 72 hours resulted in significant reduction of VEGF levels (down to 396.66 +/- 257.47 pg/ml, 363.42 +/- 239.85 pg/ml and 407.47 +/- 219.38 pg/ml, respectively) (P < 0.05). In conclusion, abnormal high expression of VEGF plays a role in the pathogenetic course of CML and it is probably an additional anticancer mechanism for As(2)O(3) to inhibit VEGF expression of leukemic cells.
Adolescent ; Adult ; Aged ; Arsenicals ; pharmacology ; Bone Marrow Cells ; drug effects ; metabolism ; Cells, Cultured ; Child ; Culture Media, Conditioned ; chemistry ; Down-Regulation ; drug effects ; Endothelial Growth Factors ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; pathology ; Lymphokines ; metabolism ; Male ; Middle Aged ; Oxides ; pharmacology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Notch signal path plays important roles in the regulation of proliferation and differentiation of hematopoietic stem cells. An extracellular domain of human Delta-like-1 (hDll-1(ext)), one of Notch ligands, was cloned and expressed in CHO cells, and the effect of hDll-1(ext) on expansion of hematopoietic stem/progenitor cells was investigated in this study. Total RNA was isolated from human marrow mononuclear cells. hDll-1(ext) was amplified by RT-PCR and cloned to T vector, then the gene was sequenced and subcloned to pcDNA3.1/Myc-His(+)A expression vector. The constructed plasmid was transfected into CHO cells with lipofectin and the expression of secreted hDll-1(ext) in G418-resistant clones was assayed by Western blot. hDll-1(ext) high-expressed clone was cultured to collect supernatant. Fusion protein hDll-1(ext) was purified from the supernatant by immobilized metal affinity chromatography (IMAC). The results showed that expression of Notch-1 receptor was detected in cord blood-derived CD34(+) cells by RT-PCR. Human umbilical blood CD34(+) cells were cultured in serum-free medium containing SCF, IL-3, VEGF, and with or without purified hDll-1(ext) for 4 or 8 days. Effect of hDll-1(ext) on the expansion of progenitor cells was analyzed then by clonogenic assays. The number of CFU-Mix and HPP-CFC generated from the culture system containing hDll-1(ext) was 1.5 times of that from the control. In conclusion, the recombinant hDll-1(ext) promotes the expansion of primitive hematopoietic progenitors.
Animals ; Antigens, CD34 ; immunology ; Binding Sites ; genetics ; CHO Cells ; Cell Division ; drug effects ; physiology ; Colony-Forming Units Assay ; Cricetinae ; Endothelial Growth Factors ; pharmacology ; Fetal Blood ; cytology ; immunology ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Glycoproteins ; genetics ; pharmacology ; physiology ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Interleukin-3 ; pharmacology ; Lymphokines ; pharmacology ; Membrane Proteins ; genetics ; RNA ; genetics ; metabolism ; Receptor, Notch1 ; Receptors, Cell Surface ; Recombinant Proteins ; isolation & purification ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; pharmacology ; Transcription Factors ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVESTo determine the pre-therapeutic serum level of vascular endothelial growth factor (VEGF) in patients with hepatocellular carcinoma (HCC) and to elucidate the relation between the serum level and clinical characteristics and metastasis of HCC.

METHODSOne-hundred and fifteen HCC patients, 40 patients with benign liver lesions, and 30 healthy control subjects were included in this study. The serum VEGF level was measured with the quantitative sandwich enzyme linked immunosorbent assay (ELISA, R&D systems).

RESULTSThe serum VEGF levels in the HCC group (465.62 +/- 336.24 pg/ml) was significantly elevated as compared with those in patients with benign liver lesions (159.54 +/- 120.58 pg/ml) and those in normal controls (123.53 +/- 51.84 pg/ml). The VEGF levels were not significantly different between the patients with benign liver lesions and the normal controls. The serum VEGF levels showed a positive rate of 77.4%, 25%, and 3.3% in the HCC patients, benign liver lesion patients and normal controls, respectively. In the 115 HCC patients, the serum VEGF levels in patients with portal vein (PV) emboli (n = 26, 582.76 +/- 441.89 pg/ml), with metastasis (n = 43, 548.29 +/- 438.57 pg/ml) or with large HCC lesions (>/= 5 cm in diameter) (n = 69, 554.43 +/- 369.99 pg/ml) were significantly higher than those without PV-emboli (n = 89, 431.39 +/- 292.84 pg/ml), without metastasis (n = 72, 416.24 +/- 247.27 pg/ml) or with small HCC lesions (n = 42, 328.67 +/- 227.47 pg/ml). The serum VEGF levels in stage I, II, III, IVa and IVb HCC patients were 340.6 pg/ml, 451.55 +/- 307.84 pg/ml, 397.44 +/- 257.18 pg/ml, 486.10 +/- 397.73 pg/ml and 647.93 +/- 344.56 pg/ml, respectively.

CONCLUSIONThe pre-therapeutic serum VEGF levels in HCC patients appear to reflect the disease's potential activity of vascular invasion and metastasis.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; pathology ; Endothelial Growth Factors ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; Liver Neoplasms ; blood ; pathology ; Lymphokines ; blood ; Male ; Middle Aged ; Neoplasm Metastasis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVELooking for an objective biomedical index to distinguish types and phases of hemangioma in order to provide an objective basis for selecting clinical treatment to hemangioma.

METHODSELISA (enzyme-linked immunosorbent assay) was used to determine serum VEGF concentration of 15 patients with proliferative hemangioma, 6 with involuted hemangioma, 6 with vascular malformation and 8 infants of the control group.

RESULTSThe serum VEGF concentrations of 15 proliferative hemangioma patients were significantly higher than those of involuted hemangioma patients, vascular malformation patients and control group infants. The serum VEGF concentrations of involuted hemangioma patients were a little bit higher than those of vascular malformation patients and control group infants, but without statistic significance.

CONCLUSIONSELISA could easily and accurately determine the serum VEGF concentration of different types and different phases of hemangioma. The determination of serum VEGF concentration could provide guidance for selecting a protocol of systemic corticosteroid treatment for proliferative hemangioma. Combined with gene expression and distribution of VEGF and its receptors and some other cytokines, the determination of serum VEGF concentration could help elucidate the mechanism of proliferative hemangioma.

Endothelial Growth Factors ; blood ; Enzyme-Linked Immunosorbent Assay ; Hemangioma ; blood ; Humans ; Infant ; Lymphokines ; blood ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the pro-angiogenic effects of several multiple myeloma (MM) cell line culture supernatants on human bone marrow endothelial cell (HBMEC) proliferation, migration, and capillary formation, and the anti-angiogenic effects of thalidomide.

METHODSHBMEC was cultured in the presence of MM cell lines (IM9, XG1, U266 and MOLP-5) supernatants. Proliferation and migration of HBMEC were determined, capillary-like tubule formation of HBMEC was examined in fibrin and Matrigel. The inhibiting effect of thalidomide was investigated by adding it into myeloma cell line culture supernatants. Vascular endothelial growth factor (VEGF) was measured by ELISA.

RESULTS(1) MM cell lines culture supernatants promoted HBMEC proliferation and migration. (2) In fibrin and Matrigel, capillary-like tubule network formation promoted by the supernatants. (3) All of these effects could be inhibited by thalidomide. (4) This effect was not related to VEGF in the supernatants.

CONCLUSIONSMM cell line promote proliferation, migration and tubule formation by secreting VEGF or other several cytokines. Thalidomide can inhibit these effects.

Angiogenesis Inhibitors ; pharmacology ; Bone Marrow ; blood supply ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; chemistry ; pharmacology ; Endothelial Growth Factors ; metabolism ; Endothelium, Vascular ; cytology ; drug effects ; physiology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; metabolism ; Multiple Myeloma ; pathology ; secretion ; Neovascularization, Physiologic ; drug effects ; Thalidomide ; pharmacology ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo observe the effective mechanism and side effects of thalidomide to multiple myeloma (MM).

METHODSTen cases of MM were studied, of which 3 were previously untreated and 7 refractory or relapsed. Bone marrow microvascular density (MVD) was detected by factor-VIII related antigen and CD(34) immunohistological staining and serum concentration of vascular endothelial growth factor (VEGF) before and after treatment was determined by ELISA. The initial dosage of thalidomide was 100 approximately 200 mg/d with a weekly escalation of 50 mg/d to 450 approximately 650 mg/d. The therapeutic effectiveness is classified into partial remission, improvement and uneffective according to the decrease of serum M protein and bone marrow myeloma cells. Anemia, renal function and blood electrolytes were also observed.

RESULTSBefore treatment, MVD was 73.32 +/- 28.80 and 32.30 +/- 12.50 in MM and control group, respectively, (P < 0.01). MVD in MM group decreased to 56.12 +/- 19.34 after treatment, and was of significant difference (P < 0.05) as compared to the pretreatment value. However, there was still a significant difference as compared to control (56.12 +/- 19.34 vs 32.30 +/- 12.50, P < 0.01). The concentration of VEGF significantly decreased after treatment [from (178.23 +/- 26.56) ng/L to (78.48 +/- 19.98) ng/L, P < 0.01)]. The total effective rate was 70%. There were no serious side effects.

CONCLUSIONMVD and VEGF concentration were decreased obviously by thalidomide treatment. The dosage of 450 approximately 650 mg/d might be effective in refractory or initial MM.

Aged ; Angiogenesis Inhibitors ; adverse effects ; therapeutic use ; Antigens, CD34 ; analysis ; Bone Marrow ; blood supply ; drug effects ; Constipation ; chemically induced ; Endothelial Growth Factors ; blood ; Fatigue ; chemically induced ; Female ; Humans ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; blood ; Lymphokines ; blood ; drug effects ; Male ; Middle Aged ; Multiple Myeloma ; blood ; drug therapy ; pathology ; Nausea ; chemically induced ; Sleep Wake Disorders ; chemically induced ; Thalidomide ; adverse effects ; therapeutic use ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors ; von Willebrand Factor ; 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

OBJECTIVESTo investigate mechanism for the increasing level of serum vascular endothelial growth factor(VEGF) in tumour patients during radiotherapy and the inhibitory action of the antisense oligodeoxynucleotide (AS-ODN) to the expression of VEGF protein by radiotherapy in the prostate cancer cell line (PC3M).

METHODSTo observe the changes of serum VEGF in the prostate cancer patients during radiotherapy dynamically and the inhibitory action of the antisense oligodeoxynucleotide to the expression of VEGF by radiotherapy in PC3M.

RESULTSThe changes of serum VEGF in three patients receiving radiotherapy had been observed continuously. The levels of serum VEGF began to increase when the patients received radiotherapy and rised up to peak value after fifteen days, then declined to the range of pre-radiotherapy. Irradiating the PC3M cells with X-rays significantly increased the VEGF expression and secretion. The expression of VEGF protein in the group treated by VEGF AS-ODNs and X-ray irradiation decreased significantly than the group treated only by X-ray irradiation.

CONCLUSIONSThe induction of VEGF protein expression by X-ray irradiation in tumor cells may result in the increasing of the VEGF in the prostate cancer patients during radiotherapy and the induction can be blocked by VEGF AS-ODNs.

DNA, Antisense ; pharmacology ; Endothelial Growth Factors ; antagonists & inhibitors ; blood ; genetics ; Gene Expression ; drug effects ; radiation effects ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; genetics ; Lymphokines ; antagonists & inhibitors ; blood ; genetics ; Male ; Prostatic Neoplasms ; blood ; pathology ; Radiotherapy ; adverse effects ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo detect the biological factor association with metastasis and recurrence of hepato cellular carcinoma (HCC).

METHODSLabeled streptavidin biotin method was performed to study VEGF, uPA and ICAM-1 protein, and antigen of PCNA expression in 123 patients with HCC. Venous invasion was observed under microscope at the same time.

RESULTSThe expression rate of VEGF was higher in HCC with intra-hepatic metastasis in group B than in HCC without PVTT/metastasis in group A (P < 0.01) and higher in HCC with PVTT in group C and PVTT in group D higher than in group B (P < 0.05). The expression rate of uPA protein was higher in group B than in group A (P < 0.01), but no significant difference in groups B and C. The expression rate of ICAM-1 showed no significant difference in the four groups. MVD and PCNA-LI increased gradually from group A to D. The rate of microscopic venous invasion in group B was higher than in group A (P < 0.05), in group D higher than in group B (P < 0.05), but no significant difference was observed between groups B and C, groups C and D (P = 0.16, 0.13 respectively). The rate of postoperative recurrence of HCC was higher in group B than in group A, and lower than in group C. Multivariate regression analysis: showed postoperative recurrence was correlated very well with microscopic venous invasion (r = 0.783, P < 0.01), and MVD (r = 0.143, P < 0.05). Metastasis of HCC were associated very well with PCNA-LI (r = 0.590, P < 0.01) and MVD (r = 0.179, P < 0.05), and negatively correlated with the rate of ICAM-1 expression (r = -0.183, P < 0.01).

CONCLUSIONSVEGF, uPA and ICAM-1 protein expression and proliferation of cancer cells could contribute to the formation of PVTT, metastasis and postoperative recurrence of HCC. Over-proliferated cancer cells in HCC could be the direct factor of intrahepatic metastasis and formation of PVTT, and microscopic venous invasion may be a significant factor to predict postoperative recurrence of HCC.

Carcinoma, Hepatocellular ; blood supply ; pathology ; secondary ; Endothelial Growth Factors ; analysis ; Humans ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; analysis ; Intercellular Signaling Peptides and Proteins ; analysis ; Liver Neoplasms ; blood supply ; chemistry ; pathology ; Lymphokines ; analysis ; Neoplasm Recurrence, Local ; Proliferating Cell Nuclear Antigen ; analysis ; Urokinase-Type Plasminogen Activator ; analysis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors