Differential expression of hypoxia-inducible factor 1α in non-small cell lung cancer and small cell lung cancer

OBJECTIVES:

The aim of this study was to compare the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor in small cell lung cancer and subtypes of non-small cell lung cancer and examine their relationships with clinicopathologic factors, response to treatment and survival.

METHODS:

We examined samples obtained by bronchial endoscopic biopsy from 55 patients with inoperable lung cancer (16 with adenocarcinoma, 17 with squamous cell carcinoma, and 22 with small cell lung cancer). Hypoxia-inducible factor 1α and vascular endothelial growth factor were detected using immunohistochemistry. The diagnosis, treatment, and follow-up of patients were conducted according to the standard practice.

RESULTS:

A significant difference (p = 0.022) in hypoxia-inducible factor 1α expression was observed between non-small cell lung cancer (75.8% positive) and small cell lung cancer (45.5% positive). The frequency of hypoxia-inducible factor 1α nuclear expression was 88.2% in squamous cell carcinoma, 62.5% in adenocarcinoma, and 45.5% in small cell lung cancer. A significant correlation was observed between hypoxia-inducible factor 1α and vascular endothelial growth factor expression (Fisher''s exact test, p = 0.001) when all types of lung cancer were examined, either collectively or separately.

CONCLUSIONS:

The expression of hypoxia-inducible factor-1α differs significantly between subtypes of lung cancer. These findings could help elucidate the biology of the different types of non-operable lung carcinomas and have implications for the design of new therapeutic approaches for lung cancer.     

Prognostic value of c-FLIPL/s,HIF-1α, and NF-κβ in stage II and III rectal cancer

Locally advanced rectal cancer (LARC) is associated with a 25 % rate of metastases. The prognostic and predictive relevances of the expression of five proteins (c-FLIP_L/s, HIF-1α, β-catenin, p65, and p105/p50 NF-κβ) were assessed. This is a retrospective study. From 1998 to 2009, 152 patients with stage II/III rectal cancer were treated with radio-chemotherapy. TMAs constructed with tumor and normal tissue from the diagnostic endoscopic biopsy and the surgical specimen after chemoradiotherapy were subjected to immunohistochemical (IHC) analysis. Results were correlated with clinical and pathological data, including progression-free survival (PFS). Four different IHC conditions were independent prognostic parameters for PFS: (1) cytoplasmic c-FLIP_L/s (p?=?0.007), (2) nuclear HIF-1α (p?=?0.020), (3) a change in the cytoplasmic p65 between the diagnostic biopsy and the post-treatment specimen (p?=?0.004), and (4) a change in the cytoplasmic c-FLIP_L/s between the diagnostic biopsy and the post-treatment specimen (p?=?0.021). Three different protein expression profiles, combining biomarkers, showed prognostic significance. IHC evaluation of these biomarkers in our three protein expression profiles may help to identify patients with worse prognosis and design more effective therapeutic strategies to personalize the treatment of rectal cancer.     

The distribution pattern of ERα expression,ESR1 genetic variation and expression of growth factor receptors: association with breast cancer prognosis in Russian patients treated with adjuvant tamoxifen

Identification of additional biomarkers associated with ER genomic and nongenomic pathways could be very useful to distinguish patients who will benefit from tamoxifen treatment. The aim of this study was to analyze the prognostic significance of the distribution pattern of ERα expression, ESR1 gene single-nucleotide polymorphisms and expression levels of growth factor receptors in Russian hormone receptor-positive breast cancer patients treated with adjuvant tamoxifen. Formalin-fixed paraffin-embedded tumor tissue samples from 97 patients were examined for the distribution pattern of ERα expression, as well as for EGFR and TGF-βR1 expression by immunohistochemistry. Genotypes for ESR1 +30T>C (rs2077647) and ESR1 2014G>A (rs2228480) were analyzed using a TaqMan assay. Progression-free survival (PFS) was used as an endpoint for the survival analyses. We found that patients with the heterogeneous distribution of ERα expression had poor prognosis on tamoxifen treatment (P = 0.021). We identified a high EGFR expression in patients who developed distant metastasis or recurrence during tamoxifen treatment (a tamoxifen-resistant group—TR) in contrast to the distant metastasis-free patients (a tamoxifen-sensitive group—TS) (80.0 vs. 41.9 %, respectively, P = 0.009). Carriers of the ESR12014A mutant allele were more prevalent among the TR patients compared to the TS patients (26.3 vs. 8.0 %, respectively, P = 0.009). EGFR expression and the distribution pattern of ERα expression were associated with the response to tamoxifen by both univariate and multivariate logistic regression analyses. The presence of these markers either alone or in combination was correlated with the worse PFS for all patients. Analysis of the distribution pattern of ERα expression and the EGFR status in tumor tissue may be valuable for patient selection for tamoxifen adjuvant therapy.     

Tanshinone IIA-induced attenuation of lung injury in endotoxemic mice is associated with reduction of hypoxia-inducible factor 1α expression

Inhibiting hypoxia-inducible factor (HIF)-1α activity has been proposed as a novel therapeutic target in LPS-induced sepsis syndrome. We have reported that tanshinone IIA (TIIA) can reduce LPS-induced lethality and lung injury in mice, but the precise mechanisms have not been fully described. Therefore, the present study investigated whether the protective effect of TIIA was related to the inhibition of LPS-induced HIF-1α expression and what mechanisms accounted for it. This study showed that TIIA pretreatment improved LPS-induced biochemical and cellular changes and reduced the production of inflammatory cytokines. Pretreatment with TIIA decreased LPS-induced HIF-1α expression in vivo and in vitro. TIIA did not affect the LPS-induced HIF-1α mRNA level but inhibited HIF-1α protein translation by the inhibition of the PI3K/AKT and MAPK pathways and related protein translational regulators, such as p70S6K1, S6 ribosomal protein, 4E-BP1, and eIF4E, and promoted HIF-1α protein degradation via the proteasomal pathway in LPS-stimulated macrophages. These observations partially explain the antiinflammatory effects of TIIA, which provides scientific basis for its application for the treatment of acute lung injury/acute respiratory distress syndrome or sepsis.     

High altitude increases the expression of hypoxia-inducible factor 1α and inducible nitric oxide synthase with intest-inal mucosal barrier failure in rats

Background: The failure of intestinal mucosal barrier may induce multiple organ dysfunction and systemic inflammatory response syndrome, but little work has been done on whether hypobaric hypoxia related to the failure of intestinal mucosal barrier. Aims: To study the expression of hypoxia-inducible factor 1α (HIF-1α), inducible nitric oxide synthase (iNOS) and morphological changes of intestinal mucosa in albino rats at different altitude. Methods: 30 male Wistar rats raised in plain for one month were randomly divided into 3 groups: Plain 500 m group (n=10), High-altitude (HA) 3842 m group (n=10) and HA4767 m group (n=10). Each group was delivered to different altitude area at the same shipping time and executed after 3 days’ exposure to different altitude. Intestinal segments with the same location of all rats were removed for morphological analyses. Morphologic parameters (villous height, crypt depth, mucosal wall thickness and villous surface area) were measured by optical and scanning electron microscope. The expression of iNOS and HIF-1α were detected by immunohistochemistry. Results: Morphological indexes in higher altitude groups were exacerbated obviously compared with those of lower altitude groups. While the expression of iNOS and HIF-1α in higher altitude groups were significantly increased than those of lower altitude groups. Linear correlation analysis showed that the expression of iNOS was positively correlated with that of HIF-1α. Conclusions: Hypobaric hypoxia increases the expression of HIF-1α and iNOS in intestinal mucosa, however exacerbates the mucous morphologic parameters with altitude increasing. HIF-1α may regulate the expression of iNOS and be involved in the damage of intestinal mucosa.     

Expression of estrogen receptors-α and -β in primary breast neoplasms and tumors exposed to neoadjuvant hormonal therapy

We studied the content and expression of mRNA for estrogen receptors receptors- and - in breast tumors before and after 3-month neoadjuvant hormone therapy with antiestrogen tamoxifen and/or aromatase inhibitors. Expression of estrogen receptors- and - was most often detected in ER⁺PR⁺ tumors and most significantly decreased in these neoplasms after exemestane therapy. Immunocytochemical and radioligand assays showed that tamoxifen and anastrozole have little effect on the number of estrogen receptors- The number of progesterone receptors in tumors decreased by the end of anastrozole therapy. Estrogen receptors- were immunocytochemically revealed in 50% primary breast tumors. Anastrozole slightly decreased, while tamoxifen increased the incidence of these receptors. Interruption of signaling through estrogen receptors and suppression of estrogen biosynthesis had different effects on the receptor status of neoplasms and distribution of estrogen receptors- and -.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 11, pp. 559–562, November, 2004     

Immunohistochemical expression HIF1α in chronic plaque psoriasis,an association with angiogenesis and proliferation

Psoriasis is characterized by excessive cell proliferation, angiogenesis, and regions of hypoxia. Hypoxia stimulates production of hypoxia inducible factors (HIFs) such as HIF1α. The aim of the present study is to investigate the possible role of HIF1α in pathogenesis of psoriasis and to correlate its expression with angiogenesis and proliferation in involved and uninvolved skin in patients with plaque psoriasis using CD34 and Ki-67. The current study was performed on 40 skin specimens of patients presented with chronic plaque psoriasis both involved and uninvolved together with 40 specimens from age- and sex-matched healthy volunteers as a control group. The specimens were submitted for HIF1α, CD34, and Ki-67 immunostaining. HIF1α was expressed in 37.5% of normal skin with mild intensity and cytoplasmic localization instead of its expression in 72.5% and 100% of uninvolved and involved psoriatic skin, respectively. Nucleocytoplasmic pattern of HIF1α was seen in 34.5% and 37.5% of uninvolved and involved psoriatic skin, respectively. Positive and intense expression of HIF1α as well as its nucleocytoplasmic localization were significantly in favor of psoriatic skin either involved or uninvolved in comparison to normal skin (P < 0.05). Intense HIF1α was significantly associated with microvessel density in both involved and uninvolved skin (P < 0.05). Nucleocytoplasmic pattern was significantly associated with epidermal acanthosis (P < 0.05) and tended to be associated with percentage of Ki-67 of psoriatic skin (P = 0.06). The present study demonstrated that HIF1α is upregulated in the skin of psoriatic cases (involved and uninvolved) compared to normal skin indicating its role in pathogenesis of psoriasis especially its active nuclear form that showed an association with angiogenesis and proliferation.     

Coexpression of hypoxia-inducible factor-1α and glucose transporter-1 is associated with poor prognosis in oral squamous cell carcinoma patients

Eckert A W, Lautner M H W, Schütze A, Taubert H, Schubert J & Bilkenroth U (2011) Histopathology 58 , 1136–1147 Coexpression of hypoxia‐inducible factor‐1α and glucose transporter‐1 is associated with poor prognosis in oral squamous cell carcinoma patients Aims: To study whether coexpression of the two hypoxia‐related proteins hypoxia‐inducible factor (HIF)‐1α and glucose transporter (GLUT)‐1 has prognostic relevance in oral squamous cell carcinomas (OSCCs). Methods and results: Eighty‐two OSCC samples were analysed for expression levels of HIF‐1α and GLUT‐1 by immunohistochemistry. Protein expression was assessed with an immunoreactive score system, and the correlations between gene expression and both clinical and pathohistological parameters were examined. Overexpression of either GLUT‐1 or HIF‐1α was associated with poor disease‐specific survival in OSCC patients. Multivariate Cox proportional‐hazards regression analysis revealed that increased expression of HIF‐1α was significantly associated with disease‐specific survival (relative risk = 3.24, P = 0.024), as compared with the group with a low level of expression. Coexpression of HIF‐1α and GLUT‐1 was additively and significantly associated with adverse prognoses in patients with OSCC. Patients whose tumours had increased levels of expression of both HIF‐1α and GLUT‐1 were found to have a 5.13‐fold increased risk of tumour‐related death (P = 0.017). Conclusions: Coexpression of high levels of HIF‐1α and GLUT‐1 is significantly correlated with prognosis in OSCC patients, suggesting that the coexpression of these proteins can be used as both an early diagnostic and independent prognostic marker.     

Angiogenin and SDF-1α serum concentration in patients with systemic sclerosis in relation to clinical status

Introduction

Systemic sclerosis (SSc) is a connective tissue disorder characterized by tissue hypoxia due to vascular changes and excessive fibrosis of the skin and internal organs. Damage to blood vessels and endothelium, as well as imbalance of vascular homeostasis, impairment of angiogenesis and vasculogenesis are observed in the course of the disease. The aim of the study was to investigate the pro-angiogenic factors angiogenin and SDF-1α in patients with SSc.

Material and methods

Serum samples were collected from 50 patients with dSSc (diffuse SSc) and lSSc (limited SSc) and from 38 patients used as a healthy control group. We explored: 1) how the serum concentrations of SDF-1α and angiogenin differ in the investigated groups; 2) the correlation among chemokines in SSc and the duration of the disease, Raynaud’s phenomenon, sclerosis of the skin and TSS (total skin score).

Results

Patients with SSc showed statistically significantly higher serum angiogenin concentration and there was no correlation between duration of the disease and Raynaud’s phenomenon, skin sclerosis or TSS. There was also no difference or no correlation between serum level of SDF-1α and the investigated groups.

Conclusions

The increase in angiogenin concentration in the serum in patients with SSc may confirm endothelial damage caused by hypoxia and reduced vascular perfusion due to the course of SSc without contributing to compensatory revascularization.     

Is reduced expression of mismatch repair genes MLH1 and MSH2 in patients with sporadic colorectal cancer related to their prognosis?

The majority of mutations in hereditary nonpolyposis colon carcinoma (HNPCC) patients affect the mismatch-repair genes (MMRG) MLH1 and MSH2. In addition, mutations of these genes were found in about 15% of sporadic colorectal carcinomas which appear to be related to microsatellite instability (MSI). However, mutations in MMRG were not found in all MSI-positive carcinomas, but MMRG mutations may be relevant for the assessment of tumor characteristics and patients’ prognosis. Therefore, we investigated the relationship between expression of MMRG, tumor biology and patients’ survival. In 127 patients with sporadic colorectal carcinomas and a minimum of 5 years follow-up after curative surgery immunohistochemical detection of MLH1 and MSH2 was analyzed semiquantitatively. Lost expression of MLH1 has been found in tumor specimens from 10 patients, whereas MSH2 expression was missing in 5 patients. This reduced expression did not correlate with tumor stage, lymph node involvement, grading or tumor invasion into blood vessels. However, a significant correlation was found for lymphovascular invasion (P=0.02) and localization within the colorectum (P=0.003) in MLH1-negative carcinomas. In addition, although there was a clear tendency for longer overall survival (72 vs. 63 months) for patients with MLH1-negative carcinomas, significant differences for overall and recurrence-free survival were not seen. In conclusion of our results and a critical review of literature, the prognostic importance of the MMR genes in sporadic colorectal carcinomas remains controversial.     

Polymorphisms in poly (ADP-ribose) polymerase-1 (PARP1) promoter and 3’ untranslated region and their association with PARP1 expression in breast cancer patients

Within the past several years, inhibition of the PARP1 activity has been emerged as one of the most exciting and promising strategies for triple-negative breast cancer (TNBC) therapy. The purpose of this study is to assess PARP1 expression in TNBCs and to evaluate the association between polymorphisms in PARP1 promoter or 3’ untranslated region (3’UTR) and PARP1 expression. It was found that PARP1 was overexpressed in nuclear (nPARP1), cytoplasm (cPARP1) and nuclear-cytoplasmic coexisting (coPARP1) of 187 TNBCs in comparison to that of 115 non-TNBCs (nPARP1, p<0.001; cPARP1, p<0.001; coPARP1, p<0.001). High expression of nPARP1 and cPARP1 in breast cancer was related to worse progression-free survival (nPARP1, p=0.007, cPARP1, p=0.003). Additionally, we identified seven published polymorphism sites in the promoter region and in 3’UTR of PARP1 by sequencing. rs7527192 and rs2077197 genotypes were found to be significantly associated with the cPARP1 expression in TNBC patients (rs7527192 AA+GA versus GG, p=0.014; rs2077197 AA+GA versus GG, p=0.041). These findings were confirmed in an independent validation set of 88 TNBCs (rs7527192 GG versus GA+AA, p=0.030; rs2077197 GG versus GA+AA, p=0.030). The PARP1 over-expression including nuclear, cytoplasm and nuclear-cytoplasmic coexisting is a feature of TNBCs and the assessment of its expression may help to predict the efficacy of chemotherapy with PARP1 inhibitor.     

14-3-3ζ Overexpression and abnormal β-catenin expression are associated with poor differentiation and progression in stage I non-small cell lung cancer

Recent studies have shown that 14-3-3ζ interacted with other key cellular proteins involved in the tumor development and progression. Knowledge of 14-3-3ζ and β-catenin expression and clinical significance in the same tumor tissues is limited. The purpose of this study was to investigate the expression and significance of 14-3-3ζ and β-catenin in stage I non-small-cell lung cancer (NSCLC). Specimens of NSCLC and adjacent normal lung tissues were collected from 110 patients. The expressions of 14-3-3ζ and β-catenin were detected by western blotting, double labeling immunofluorescence, confocal laser scanning microscopy and immunohistochemistry. The expression of 14-3-3ζ was upregulated in stage I NSCLC. Further, the overexpression of 14-3-3ζ correlated with histological grades, lymph node metastasis and poor clinical outcome. Abnormal expression of β-catenin was significantly correlated with poor differentiation and lymph node metastasis. Abnormal β-catenin expression was associated significantly with positive 14-3-3ζ expression. In conclusion, 14-3-3ζ and β-catenin might have an important role in development, progression and metastatic process of NSCLC. 14-3-3ζ might be used as prognostic biomarkers for NSCLC.     

Levels and patterns of expression of hypoxia-inducible factor-1α, vascular endothelial growth factor, glucose transporter-1 and CD105 in adenoid cystic carcinomas with high-grade transformation

Costa A F, Tasso M G, Mariano F V, Soares A B, Chone C T, Crespo A N, Fresno M F, Llorente J L, Suárez C, de Araújo V C, Hermsen M & Altemani A
(2012) Histopathology  60, 816–837 Levels and patterns of expression of hypoxia‐inducible factor‐1α, vascular endothelial growth factor, glucose transporter‐1 and CD105 in adenoid cystic carcinomas with high‐grade transformation Aims: To compare the expression of proteins regulated by hypoxia between adenoid cystic carcinoma (ACC) with and without high‐grade transformation (HGT). Methods and results: In eight ACC–HGT and 18 ACC without HGT, expression of hypoxia‐inducible factor‐1 (HIF‐1α), vascular endothelial growth factor (VEGF), glucose transporter‐1 (GLUT‐1) and microvascular density (MVD) by CD105 (a hypoxia‐inducible protein expressed in angiogenic endothelial cells) was determined. Expression levels of HIF‐1α and VEGF as well as CD105‐MVD did not differ significantly between: (i) transformed and conventional areas (TA and CA, respectively) of ACC–HGT, (ii) CA and ordinary ACC. HIF‐1α was detected in 100% of cases and presented a diffuse expression pattern. No significant association was found between levels of HIF‐1α expression and tumour size, metastasis and recurrence. GLUT‐1 showed a prostromal expression pattern and was observed exclusively in TA (three of six cases) and in only three of 14 ACC. Conclusions: Both the absence of significant alterations in levels of expression of HIF‐1α, VEGF and CD105 and the patterns of expression of HIF‐1α and GLUT‐1 suggest that hypoxia may not play a key role in the process of high‐grade transformation of ACC. Although HIF‐1α expression is a common finding in ACC, it cannot be used as a marker of tumour aggressiveness.     

BLCA-4 expression is related to MMP-9, VEGF, IL-1α and IL-8 in bladder cancer but not to PEDF, TNF-α or angiogenesis

Aim

BLCA-4 is a specific nuclear matrix protein found in bladder cancer and there is a dearth of study on functional analysis upon this factor. We aimed to discover whether BLCA-4 is related to angiogenesis in bladder cancer.

Methods

Fifty-three bladder cancer samples were included for immunohistochemical staining of BLCA-4, matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), interleukin-1α (IL-1α), IL-8, pigment epithelium-derived factor (PEDF), tumour necrosis factor-α (TNF-α) and von Willebrand factor (vWF) for microvessel density (MVD). Expressional levels were scored and grouped by clinicopathological parametres for statistical analysis for correlations.

Results

Positive correlations were identified between expression of BLCA-4 and IL-1α (p = 0.038), IL-8 (p = 0.001), VEGF (p = 0.002), and MMP-9 (p = 0.013). No correlation was found for PEDF (p = 0.182), TNF-α (p = 0.531) or MVD (p = 0.932). Positive correlations were also obtained in cases of advanced grade or stage, larger, recurrent and multiple tumours. Positive correlation between BLCA-4 and MMP-9 was also found in papillary urothelial neoplasm of low malignant potential (PUNLMP).

Conclusion

BLCA-4 may not effect pro-angiogenic pathways in bladder cancer, it can however interact with IL-1α, IL-8, VEGF and MMP-9 to enhance tumourigenesis and tumour invasiveness.