The role of ESM1 in the lipids metabolic reprogramming and angiogenesis of lung adenocarcinoma cells.

Background: Lung adenocarcinoma (LUAD) is one of the respiratory diseases with high mortality and incidence. As an important angiogenic factor, (Endothelial cell-specific molecule 1) ESM1 plays an important role in the occurrence and development of LUAD. However, the role and molecular mechanism of ESM1 on LUAD metabolic reprogramming and angiogenesis remain unclear. Methods: We used multiple databases to analyze the prognostic significance and potential function of ESM1 in patients with LUAD. The expression of ESM1 in LUAD cells was down-regulated/overexpressed by RNA interference, and the effects of ESM1 on the proliferation, migration, lipid metabolism and angiogenesis of LUAD cells in vitro and in vivo were analyzed using MTT, EdU, wound healing, oil red O, tubule formation, xenograft tumor model and chicken embryo allantoic model. Results: ESM1 is closely associated with poor prognosis in LUAD patients. ESM1 promotes LUAD proliferation, migration, fatty acid synthesis and angiogenesis. It also accelerates the proliferation, migration, lipid synthesis and tubule formation of endothelial cells in the tumor microenvironment in the form of secreted protein. Mechanically, ESM1 can promote the activation of AKT signaling pathway and up-regulate the expression of SCD1 and FASN. Conclusion: Our results suggest that ESM1 promotes the proliferation, migration, lipid reprogramming, and angiogenesis of LUAD cells by activating the AKT signaling pathway, suggesting that ESM1 may be a potential therapeutic target and prognostic marker in LUAD patients.

ADAM12 promotes the resistance of lung adenocarcinoma cells to EGFR-TKI and regulates the immune microenvironment by activating PI3K/Akt/mTOR and RAS signaling pathways.

Lung adenocarcinoma (LUAD) is a malignant respiratory disease, resulting in a heavy social burden. Epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) resistance and tumor immune microenvironment are important directions in the treatment of LUAD. In this study, we confirmed the role of ADAM metallopeptidase domain 12 (ADAM12) in LUAD development and progression. Our bioinformatic analysis was conducted to screen ADAM12 was correlated with EGFR-TKI and immune infiltration in LUAD patients. Our results showed that the transcription and post-transcription level of ADAM12 is significantly increased in tumor samples compared to normal samples, and ADAM12 correlated with poor prognosis in LUAD patients. High level of ADAM12 accelerated the LUAD progression via promoting proliferation, cell cycle, apoptosis escaping, immune escaping, EGFR-TKI resistance, angiogenesis, invasion and migration based on experiment validation in vitro and in vivo, which could be attenuated by ADAM12 knockdown. Further mechanistic studies suggested that the PI3K/Akt/mTOR and RAS signaling pathways were activated after ADAM12 knockdown. Therefore, ADAM12 might be validated as a possible molecular therapy target and prognostic marker for patients with LUAD.

CircPTCH1 Promotes Migration in Lung Cancer by Regulating MYCN Expression Through miR-34c-5p.

BACKGROUND: The incidence rate and mortality rate of lung cancer are the highest in the world. Therefore, further studies are needed to reveal the molecular mechanism of lung cancer progression and development. Previous study demonstrated that the deregulation of circRNAs can regulate cell biological functions in tumorigenesis and development. However, the roles of circPTCH1 in lung cancer have not yet been revealed. MATERIALS AND METHODS: The expression levels of circPTCH1, miR-34c-5p, and MYCN were measured by RT-PCR in lung cancer tissues and cells; dual-luciferase reporter and RIP assay showed that circRNA served as a sponge for miRNA, and miRNA could target mRNA. In vitro, effects of si-circPTCH1 can regulate lung cancer cells' migration, invasion were detected by CCK-8 assay, wound healing assay, and transwell assay. RESULTS: Our research demonstrated that the expression of circPTCH1 was upregulated in lung cancer tissues and cell lines and increased in metastatic tissues compared to that of non-metastatic tissues. circPTCH1 sponging miR-34c-5p to target MYCN was revealed by dual-luciferase reporter and a RIP assay. In addition, the expression level of miR-34c-5p was reduced in lung cancer tumor tissues, and MYCN was significantly increased in lung cancer tumor tissues. Pearson correlation analysis showed that miR-34c-5p with circPTCH1 and MYCN had a moderately negative correlation, and there was a moderately positive correlation between circPTCH1 and MYCN. Further, cytological studies found that circPTCH1 reduced lung cancer cells' migration and invasion by targeting MYCN via miR-34c-5p. CONCLUSION: circPTCH1 plays a tumor enhancement role in lung cancer and that can effectively promote migration, invasion and EMT by targeting the miR-34c-5p/MYCN axis. circPTCH1 may be a novel potential treatment and diagnosis biomarker for lung cancer.

Misdiagnosis of multiple synchronous small bowel adenocarcinomas as intestinal tuberculosis: a case report.

BACKGROUND: Small bowel adenocarcinoma (SBA) is a rare malignancy that primarily occurs in the duodenum. Multiple synchronous SBA is unique rare and difficult to diagnose due to non-specific disease presentation. Protocols to identify multiple synchronous SBA during early disease stages are urgently required. CASE PRESENTATION: An elderly man experienced left lower abdominal pain and melena for 3 months. Abdominal CT showed thickening of the multiple segmental small intestinal walls. As the patient had pulmonary tuberculosis simultaneously, he was misdiagnosis as intestinal tuberculosis and received anti-spasm therapy. The treatment delayed radical resection surgery and the patient underwent palliative segmental resection of the jejunum after 4 months due to intestinal obstruction. Resected specimens showed multiple synchronous SBA (five tumors). The patient accepted chemotherapy postoperatively. Six months postoperatively, the patient died of brain metastasis. CONCLUSIONS: We highlight how multiple synchronous SBA is rare and easily misdiagnosed. We should rule out multiple synchronous SBA when diagnosing intestinal diseases (e.g. inflammatory bowel disease, IBS). Intestinal tuberculosis may also be one of the risk factors for multiple synchronous SBA. High-risk patients should be assessed for known tumor makers, and receive gastroscopy, enteroscopy or capsule endoscopy. Doctors should obtain the pathology under endoscopy to the greatest possible degree. For suspected patients, laparotomy should be performed.

Knockdown of Long Non-Coding RNA (lncRNA) Colon Cancer-Associated Transcript-1 (CCAT1) Suppresses Oral Squamous Cell Carcinoma Proliferation, Invasion, and Migration by Inhibiting the Discoidin Domain Receptor 2 (DDR2)/ERK/AKT Axis.

BACKGROUND Emerging evidence shows that lncRNAs are involved in carcinogenesis or suppression in diverse cancers. This study assessed the biological role of lncRNA CCAT1 in OSCC and explored the underlying molecule mechanism. MATERIAL AND METHODS CCAT1 and DDR2 expression was measured by qRT-PCR. Colony formation assay and CCK-8 assay were performed to evaluate cell proliferation. Cell cycle was determined by flow cytometric analysis and Western blot analysis. In addition, wound healing and Transwell assay were used to assess cell migration and invasion, respectively. RNA immunoprecipitation (RIP) assay were employed to identify the interaction between DDR2 and CCAT1. Protein levels involved in DDR2/ERK/AKT pathway were estimated by Western blot assay. RESULTS The findings revealed that CCAT1expression was upregulated in OSCC cell lines. Knockdown of CCAT1 repressed cell proliferation, blocked the cell cycle, and suppressed the invasion and migration of TCA-8113 cells. Moreover, DDR2 expression in OSCC cell lines was downregulated and CCAT1 silencing repressed the expression of DDR2. RIP assays validated the binding of CCAT1 and DDR2 protein. Moreover, CCAT1 silencing suppressed the ERK/AKT signaling through DDR2 in TCA-8113 cells. CONCLUSIONS Downregulation of CCAT1 suppressed TCA-8113 cell proliferation, invasion, and migration by inactivation of the ERK/AKT pathway via inhibition of DDR2, suggesting the value of CCAT1 in diagnosis and treatment of patients with OSCC.

A large basal cell adenoma extending to the ipsilateral skull base and mastoid in the right parotid gland: A case report.

BACKGROUND: Basal cell adenoma (BCA) is a rare benign tumour that has unique histological characteristics and primarily arises in the parotid glands. According to published reports, nearby tissue destruction by BCA seems impossible. CASE SUMMARY: We presented a case of a 54-year-old woman with a mass in the deep lobe of the right parotid gland involving the ipsilateral skull base and mastoid. The patient exhibited gradual right facial swelling but no other obvious symptoms. Combined resection of the total right parotid gland and partial skull base excision were performed. The biopsy conducted before the surgery and sections cut from intraoperatively obtained tissues were not definitive for identifying the character of the neoplasm. A final diagnosis of tubular BCA without malignant elements was established based on postoperative pathology results and immunohistochemical analysis. The tumour did not recur during the 12-mo follow-up period. CONCLUSION: A diagnosis of BCA can only be established based on a histopathological examination after an excisional biopsy, and tubular BCA should carefully be considered as a destructive type.

Activation of PINK1-Parkin-dependent mitophagy in Tri-ortho-cresyl phosphate-treated Neuro2a cells.

Tri-ortho-cresyl phosphate (TOCP) is a typical organophosphorus compound that can cause organophosphate-induced delayed neuropathy (OPIDN), which is pathologically characterized by axonal degeneration. Nowadays, mitochondrial dysfunction is regarded as a potential mechanism contributing to OPIDN progress. Mitophagy, a selective type of autophagy, is required to segregate damaged mitochondria from healthy mitochondrial networks and deliver them to lysosome for degradation. This research was designed to investigate the role of mitophagy in axon degeneration following TOCP administration in an in vitro model. Differentiated neuro2a (N2a) cells were divided into four groups and treated with 0, 5, 10, and 20 µM TOCP for 24 h, respectively. The critical proteins in PINK1-Parkin-dependent mitophagy including LC3, P62, PINK1, Parkin, mitochondrial proteins, and autophagic receptors were detected by immunoblotting and immunofluorescence. After TOCP treatment, increased level of ROS in N2a cells revealed a significant mitochondria damage. Meanwhile, it was observed that much more PINK1, Parkin, and LC3-II were translocated to the mitochondria. Furthermore, immunofluorescence analysis demonstrated that the co-localization of Parkin and LC3 was significantly increased. These results suggested that PINK1-Parkin dependent mitophagy pathway in N2a cells was activated by TOCP treatment. In addition, P62, a major autophagic receptor, was markedly accumulated on the mitochondria, which indicated that P62 might play a critical role in facilitating mitophagy under TOCP-induced axonal degeneration. Taken together, our results suggest that TOCP exposure resulted in the activation of PINK1-Parkin-dependent mitophagy in N2a cells. Mitophagy may act as a positively reactive mode in eliminating dysfunctional mitochondria and therefore protect neurons against TOCP neurotoxicity.

Evolution of Dislocation Loops Induced by Different Hydrogen Irradiation Conditions in Reduced-Activation Martensitic Steel.

Hydrogen can be induced in various ways into reduced-activation ferritic/martensitic (RAFM) steels when they are used as structural materials for advanced nuclear systems. However, because of the fast diffusion of hydrogen in metals, the effect of hydrogen on the evolution of irradiation-induced defects was almost neglected. In the present work, the effect of hydrogen on the evolution of dislocation loops was investigated using a transmission electron microscope. Specimens of reduced-activation ferritic/martensitic (RAFM) steels were irradiated with hydrogen ions to 5 × 1020 H⁺ • m-2 at 523⁻823 K, and to 1 × 1020 H⁺ • m-2 - 5 × 1020 H⁺ • m-2 at 723 K. The experimental results reveal that there is an optimum temperature for dislocation loop growth, which is ~723 K, and it is greater than the reported values for neutron irradiations. Surprisingly, the sizes of the loops produced by hydrogen ions, namely, 93 nm and 286 nm for the mean and maximum value, respectively, at the peak dose of 0.16 dpa under 723 K, are much larger than that produced by neutrons and heavy ions at the same damage level and temperature. The results indicate that hydrogen could enhance the growth of loops. Moreover, 47.3% 1 2   a0 <111> and 52.7% a0 <100> loops were observed at 523 K, but 1 2   a0 <111> loops disappeared and only a0 <100> loops existed above 623 K. Compared with the neutron and ion irradiations, the presence of hydrogen promoted the formation of a0 <100> loops.

Investigation of Helium Behavior in RAFM Steel by Positron Annihilation Doppler Broadening and Thermal Desorption Spectroscopy.

The behavior of helium in reduced-activation ferritic/martensitic steels was investigated systematically with positron annihilation Doppler broadening measurement and thermal desorption spectroscopy. Specimens were irradiated with helium ions with different energies to various fluences at different temperatures. A threshold fluence was observed above which the rate of formation and growth of helium bubbles dramatically increased. Irradiation at higher temperature could suppress the formation and growth of HenVm clusters with low binding energies and enhance that of helium bubbles and HenVm clusters with high binding energies. Different changes of S parameters were observed in various depth after the irradiation temperature was increased from 523 K to 723 K. Irradiation of 18 keV-He⁺ enhanced the growth of HenVm clusters and helium bubbles compared with 100 keV-He⁺ irradiation. A possible mechanism is discussed.

Screening of long non-coding RNA and TUG1 inhibits proliferation with TGF-ß induction in patients with COPD.

OBJECTIVE: To evaluate differentially expressed long noncoding RNAs (lncRNAs) and the potential role of lncRNA TUG1 in patients with chronic obstructive pulmonary disease (COPD). METHODS: Total RNA was extracted from both COPD and non-COPD lung tissues, and microarray analysis was performed with 25,628 lncRNA probes and 20,106 mRNA probes. In addition, five up-regulated and five down-regulated lncRNAs were selected for identification using quantitative real-time polymerase chain reaction. COPD cell model was established by transforming growth factor ß (TGF-ß) treatment. Cell Counting Kit-8 assay was used to detect BEAS-2B and HFL1 cell proliferation after TUG-siRNA transfection with TGF-ß treatment. In addition, the expression levels of α-SMA and fibronectin proteins were determined using Western blot in BEAS-2B and HFL1 cells after TUG-siRNA transfection with TGF-ß treatment. RESULTS: There were 8,376 (32.7%) differentially expressed lncRNAs and 5,094 (25.3%) differentially expressed mRNAs in COPD lung tissues compared with non-COPD lung tissues. Five of the analyzed lncRNAs (BC038205, BC130595, TUG1, MEG3, and LOC646329) were markedly increased, while five lncRNAs (LOC729178, PLAC2, LOC339529, LINC00229, and SNHG5) were significantly decreased in COPD lung tissues compared with non-COPD lung tissues (n=20) (***P<0.001). Knockdown of lncRNA TUG1 promotes BEAS-2B and HFL1 cell proliferation after TGF-ß treatment through inhibiting the expression levels of α-SMA and fibronectin. CONCLUSION: Abundant, differentially expressed lncRNAs and mRNAs were identified by microarray analysis and these might play a partial or key role in the diagnosis of patients with COPD. LncRNA TUG1 may become a very important class of biomarker and may act as a potential diagnostic and therapeutic target for patients with COPD.

Overexpression of HIF-1α in primary gallbladder carcinoma and its relation to vasculogenic mimicry and unfavourable prognosis.

As a novel mode of tumor neovascularization, vasculogenic mimicry (VM) has been reported to increase tumor-related mortality in many different solid tumors. In the present study, two established human gallbladder carcinoma (GBC) cell lines (highly aggressive GBC-SD and poorly aggressive SGC-996) cultured on a three-dimensional matrix were assessed for the ability of VM channel formation under normoxic or hypoxic conditions. In addition, the relationship between HIF-1α gene expression and VM channel formation of GBC cells in vitro was measured using the small interfering RNA (siRNA) technique, western blotting and real-time reverse transcription (RT)-PCR analysis. Furthermore, H&E and CD31/periodic acid-Schiff (PAS) staining were used to observe VM in GBC tissue samples. Additionally, all seventy-one specimens with VM and non-VM were stained for hypoxia inducible factor-1 α (HIF-1α) and its correlation with clinicopathological features and prognosis was analyzed simultaneously. We found that hypoxia could induce more VM channel formation and elevated HIF-1α expression in highly aggressive GBC-SD cells. HIF-1α siRNA efficiently knocked down HIF-1α expression and GBC VM networks under either normoxic or hypoxic conditions. VM was present in human primary GBC and overexpression of HIF-1α was significantly correlated with depth of invasion and perineural involvement in the non-VM group. Moreover, VM and HIF-1α were independent factors for the overall survival of GBC patients and correlated with decreased survival. In conclusion, VM was present in human GBC. As a critical mediator in VM formation, high expression of HIF-1α was associated with VM and tumor progression in GBC patients.

Chemokine receptor CXCR3 is involved in the acute pancreatitis-associated lung injury.

Acute pancreatitis is a common disease, which is divided into mild pancreatitis and severe pancreatitis. For the latter, a systemic inflammatory response may occur and lead to distant organ damage and the development of multiple organ dysfunction syndrome (MODS), which accounts for significant morbidity and mortality in humans. Chemokines and their receptors are being believed to play a pivotal role in the pathogenesis of acute pancreatitis. Chemokine receptor CXCR3 is reported to be involved in acute tissue injury, for example acute lung injury induced by cigarette smoking, but its role in acute pancreatitis is not yet known. In this study, two animal models of acute pancreatitis (cerulein- and arginine-induced pancreatitis) were applied in CXCR3⁻/⁻ mice and wild-type mice, in order to explore the role of CXCR3 in acute pancreatitis. Serum amylase, lipase and histological observations revealed that CXCR3 knockout did not affect the severity of acute pancreatitis. However, edema and inflammatory cell infiltrate in the lung tissue were attenuated in CXCR3⁻/⁻ mice when acute pancreatitis was induced. In conclusion, chemokine receptor CXCR3 is not involved in acute pancreatic injury, but has a connection with acute pancreatitis-associated lung injury. Acute pulmonary injury is attenuated in CXCR3 knockout mice in experimental acute pancreatitis.

[Over one year follow-up report for newly diagnosed pediatric Crohn's disease].

OBJECTIVE: To explore the clinical characteristics, therapeutic responses and outcomes of pediatric Crohn's disease (CD). METHODS: From a hospital-derived incidence cohort, 20 Crohn's disease patients (aged 0.5 - 15 years old at diagnosis), diagnosed from 2003 to 2009, received a follow-up period of more than one year. The patients were phenotyped according to Montreal standards. PCDAI was introduced to assess the disease activity and the Hyams J rules adopted to evaluate the therapeutic efficacies. The treatment was individualized based on the overall evaluation of child. RESULTS: Of these 20 patients, 55% were 7 - 12 years old at the diagnosis time. The male: female ratio was 1.5:1. At the time of diagnosis, the common manifestations included abdominal pain (95.0%), fever (80.0%) and diarrhea (80.0%). Growth retardation was detected in 50% of the cases. Complicated behavior was observed in 45% patients at diagnosis. The most frequent disease location at diagnosis was terminal ileum/colon (55%). Upper GI tract involvement was quite common in children (20%). Non-penetrating non-stricture (50%) behavior was most frequent at diagnosis. Ultimately, corticosteroids plus 6-MP/AZA treatment was administrated in 11 cases. Of these, 9 (82%) successfully withdrew the corticosteroids and maintained a complete remission. Colonoscopy was repeated in 6 complete remission cases and 4 of them achieved a complete mucosa healing. The mean follow-up period was 23 months (range: 12 - 59). At the endpoint of follow-up, 15 cases achieved a complete remission, 4 had a partial remission, 1 underwent operation and none of them died. The children who successfully withdrew from corticosteroids and achieved a complete remission could catch up the height of their age group. 6-MP/AZA associated severe adverse effects were reported at 17% in this group. CONCLUSION: Growth retardation is predominant in pediatric CD and it may provide diagnostic clues. Immunosuppressant therapy may improve the natural history of this disease. It is safe under close monitoring.

[Expression of tumor necrosis factor alpha mRNA in adipose cell of intrauterine growth retarded rats and its relation to insulin resistance].

OBJECTIVE: To investigate the association between the expression of turnor necrosis factor alpha (TNF-alpha) mRNA in fat tissue of intrauterine growth retarded (IUGR) rats and insulin resistance, and the long-term effects of early different nutritional diet. METHODS: The IUGR rat model was established by food restriction of pregnant rats. A total of 32 newborn IUGR rats were randomly divided into 4 groups: IUGR model (S/N) group, IUGR high caloric diet (A) group, IUGR high caloric and high protein diet (B) group, IUGR high protein diet (C) group. Only the mother rats were given those different diets individually, and all IUGR newborn pups were lactated for 3 weeks. From the beginning of the 4(th) week, all IUGR pups were weaned and fed with normal diet till the end of the experiment. Eight normal birth weight newborn rats were used as the control group fed with the normal diet. Weight, perirenal fat weight, fasting glucose and insulin concentration and quantified TNF-alpha mRNA expression in adipose cell were measured at the 48(th) week. The insulin sensitive index (ISI) and the relation index between TNF-alpha mRNA and fat weight, fat weight/body weight (fw/bw) ratio and ISI were calculated. RESULTS: ISI of IUGR model group, IUGR A and B groups was lower than normal control group, while perirenal fat weight, fw/bw and the expression of TNF-alpha mRNA in adipose cells were all significantly higher (P < 0.05 or 0.01). There were no significant differences in these indexes between IUGR C group and normal control groups (P > 0.05). A positive correlation was found between TNF-alpha mRNA and fat weight and fw/bw (r(1) = 0.755, r(2) = 0.782, P = 0.000). Significant inverse associations between ISI and TNF-alpha mRNA (r = -0.556, P = 0.000) and fw/bw (r = -0.513, P = 0.02) were also found. CONCLUSION: The occurrence of insulin resistance in IUGR rats is possibly associated with central obesity and accumulation of the abdominal fat and adipose cell over-expression of TNF-alpha. The adipose TNF-alpha may be an important pathogenic factor of insulin resistance of IUGR. High protein diet is a reasonable nutritional intervention. Because it promotes the skeleton muscle catch-up growth but not fat catch-up growth, it can avoid the occurrence of central obesity and insulin resistance in IUGR rats.

[The effects of pregnancy malnutrition on the development of insulin resistance in rat offspring].

OBJECTIVE: To investigate the effects of pregnancy malnutrition on the occurrence of insulin resistance (IR) in rat offspring during adult stage and to find out the relationship between TNF-alpha and IR; and to find out a reasonable early nutritional intervention measure for the prevention of IR, through giving different diets to offspring. METHODS: An IUGR model was built by maternal nutrition restriction. 80 newborn IUGR female pups were randomly divided into 4 groups, the mother rats were given the following diet respectively for 3 weeks after delivery, pups were fed by mother milk: (1) The IUGR (intrauterine growth retardation) rat model was used and the animals were divided into: IUGR control group (group S/N) fed with normal diet, (2) IUGR high-caloric diet group (group A), (3) IUGR high-protein and high-caloric diet group (group B) and (4) IUGR high-protein isocaloric diet group (group C). Each group had 20 pups and another 20 normal female pups were fed with normal diet as the normal control group (group C/N). All pups were weaned at the 4th week of age and fed with normal diet till the end of the experiment. At the 12th week (adulthood) and 48th week (senility) of life, body weight and length, the fasting blood glucose, insulin concentration, TNF-alpha of adipose tissue and body weight were measured. Body mass index (BMI), ISI (insulin sensitive index), IRI (insulin resistant index) and HBCI (beta cell insulin excretion index) and their correlation to TNF-alpha were calculated. RESULTS: At 12th week and 48th week of life, the insulin sensitivity of IUGR model group was significantly lower than group C/N, although there was no significant difference of body weight between these two groups. TNF-alpha was negatively correlated with ISI, positively correlated with IRI and no relation to HBCI. Group A and B was fatter and developed more severe IR. There were no significant differences in ISI, IRI, HBCI and TNF-alpha between group C and group C/N. CONCLUSIONS: IUGR offspring of pregnancy malnutrition mother rats showed IR at the age of 12th week. TNF-alpha was closely related to the occurrence of IR in IUGR pups. IUGR pups fed with high caloric diet or high protein and caloric diet at the early postnatal period amplified the metabolic abnormality. The high protein isocaloric diet is effective early nutritional intervention measure for the prevention of occurrence of IR at adulthood.

[Effect of different estrogen receptor subtypes on vascular endothelial growth factor protein expression in human breast cancer samples].

OBJECTIVES: To evaluate the relationship of vascular endothelial growth factor expression with estrogen receptor (ER) subtypes in fresh human breast cancer samples, and study the potential effect of ER subtypes on tumor angiogenesis. METHODS: Western blot was used to detect the VEGF and ERbeta protein expression in 86 fresh samples of human breast cancer. ERalpha was analyzed by immunohistochemistry routinely. RESULTS: Among 86 samples, 42 (48.8%) showed low expressed VEGF and 44 (51.2%) high expressed VEGF. The level of VEGF protein was correlated with ERbeta. In high expressed VEGF group, ERbeta was also highly expressed (chi(2) = 7.36, P < 0.01). But there was no significant difference between VEGF protein level and ERalpha (P > 0.05). CONCLUSION: In human breast cancer samples, VEGF may be related to ERbeta protein expression.