ABSTRACT
<p><b>OBJECTIVE</b>To investigate whether low molecular weight heparin (LMWH) may suppress the expression and secretion of vascular endothelial growth factor (VEGF) from tumor cells in vitro and inhibit the VEGF-induced proliferation of human tumor vascular endothelial cells.</p><p><b>METHODS</b>Human lung cancer cell line A549, human liver cancer cell line HepG2, human colon carcinoma cell lines HCT116 and HCT8 were used in this study. The expression levels of VEGF and TNF-alpha (tumor necrosis factor-alpha) in the tumor cells with or without pretreatment of LMWH/heparin were measured by standard sandwich ELISA technique. The VEGF mRNA level of HepG2 cells cultured with or without LMWH/heparin was determined by RT-PCR and real time PCR. Human umbilical vein endothelial cells (HUVEC) were cultured in tissue culture medium (TCM) with or without LMWH/heparin for 3 days. Then non-radioactive cell proliferation assay (MTS) kit and cell cycle assay by flow cytometry were performed to measure the proliferation of HUVEC.</p><p><b>RESULTS</b>The VEGF levels in the control, LMWH, and heparin groups of the pulmonary adenocarcinoma cell line A549 were (1045.89 +/- 165.30) pg/ml, (782.45 +/- 67.17) pg/ml and (916.54 +/- 71.25) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups of the colon adenocarcinoma cell line HCT116 were (955.76 +/- 51.14) pg/ml, (822.89 +/- 142.39) pg/ml and (951.77 +/- 188.22) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the colon adenocarcinoma cell line HCT8 were (1290.62 +/- 41.23) pg/ml, (1063.34 +/- 63.82) pg/ml and (1257.14 +/- 11.40) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the liver cancer cell line HepG2 were (1083.00 +/- 134.35) pg/ml, (758.00 +/- 84.85) pg/ml and (874.00 +/- 22.62) pg/ml, respectively. The VEGF expression levels in the above mentioned cell lines cultured in TCM were significantly reduced in the LMWH-treated groups compared with that of the control group (P < 0.05). But the level of TNF-alpha in TCM-cultured cells was unaffected by LMWH. The VEGF mRNA was reduced in the LMWH-treated HepG2 cell line. Moreover, TCM exhibited stimulating effect on proliferation of HUVEC and the effect was significantly impaired by LMWH treatment. Flow cytometric analysis revealed that LMWH treatment arrested HUVECs at the G1 phase of cell cycle.</p><p><b>CONCLUSION</b>LMWH can suppress the expression and secretion of VEGF by tumor cell lines and therefore have a potential inhibiting effect on angiogenesis induced by VEGF.</p>
Subject(s)
Humans , Adenocarcinoma , Metabolism , Pathology , Cell Cycle , Cell Proliferation , Cells, Cultured , Culture Media, Conditioned , Endothelial Cells , Cell Biology , HCT116 Cells , Hep G2 Cells , Heparin , Pharmacology , Heparin, Low-Molecular-Weight , Pharmacology , Lung Neoplasms , Metabolism , Pathology , RNA, Messenger , Metabolism , Tumor Necrosis Factor-alpha , Metabolism , Umbilical Veins , Cell Biology , Vascular Endothelial Growth Factor A , Genetics , Metabolism , Bodily SecretionsABSTRACT
<p><b>OBJECTIVE</b>To study the effects of phytoestrogen alpha-zearalanol (alpha-ZAL) on normal human breast.</p><p><b>METHODS</b>Ten specimens of normal human breast tissues were subcutaneously implanted into 30 athymic nude mice aged 9-10 weeks, one for 3 mice. These mice were then randomly divided into three groups: control group (without hormone treatment, n = 10), 1 mg/kg alpha-ZAL group (n = 10), and 5 mg/kg alpha-ZAL group (n = 10). All breast tissues were taken out 6 weeks later. Immunohistochemistry was used to determine the protein expressions of proliferating cell nuclear antigen (PCNA), inhibiting apoptosis gene Bcl-2, estrogen receptor (ER), and progesterone receptor (PR). Reverse transcription polymerase chain reaction (RT-PCR) was used to measure the expression levels of estrogen sulfotransferase (EST) mRNA and bridging integrator protein-1 (BIN1) mRNA. Morphological features of grafts before and after treatment were also observed.</p><p><b>RESULTS</b>Alpha-ZAL had no significant effects on Bcl-2, PCNA, ER, and PR expression of mammary epithelial cells in graft specimens. Alpha-ZAL upregulated BIN1 mRNA expression in grafts, but had no significant effect on ESTmRNA expression.</p><p><b>CONCLUSIONS</b>Alpha-ZAL does not affect the morphology, proliferating, and apoptosis of epithelial cells in normal human breast tissues implanted into nude mice, but it may increase the gene expression of tumor-inhibiting BIN1, suggesting that alpha-ZAL may have potential proteotive effect on normal human breast.</p>
Subject(s)
Adult , Animals , Female , Humans , Mice , Breast , Chemistry , Estrogens, Non-Steroidal , Pharmacology , Mice, Inbred BALB C , Mice, Nude , Phytoestrogens , Pharmacology , Proliferating Cell Nuclear Antigen , Random Allocation , Receptors, Estrogen , Receptors, Progesterone , Zeranol , PharmacologyABSTRACT
@#ObjectiveTo investigate the effect of Jiuqiang Naoliqing(JNQ)-containing serum on secretion of nitric oxide(NO) and endothelin-1(ET-1) in human umbilical vein endothelial cells(HUVECs), in order to explore the effect of JNQ on regulation of vascular active factors in HUVECs.MethodsThe HUVECs were explored to different volume fractions of JNQ containing serum after being isolated and cultured. The levels of cultured medium of NO and ET-1 were measured.ResultsThe cultured medium content of NO and ET-1 in different volume fractions of JNQ containing serum was significantly increased compared with normal serum control (P<0.05), while the ratio of NO to ET-1 were increased in comparison with normal control (P<0.05).Conclusion JNQ can be promoted secretion of NO and ET-1 in HUVECs, preserving endothelial function, and maintaining the balance of NO/E-1.