Tumour-specific arginine vasopressin promoter activation in small-cell lung cancer

Coulson, Judy M.; Stanley, Julie; Woll, Penella J.

doi:10.1038/sj.bjc.6690623

Cited by 25 publications

(30 citation statements)

References 29 publications

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“…Overexpression of USF‐2 in NSCLC stimulates cell proliferation in vitro and we have previously shown that USF‐2 is transcriptionally active in transfected NCI‐H460, where it binds on gel‐shift analysis and switches on promoter‐driven reporter gene expression for both AVP 9 and CGRP 19. In agreement with these data, the highest endogenous USF‐2 expression was observed in Lu‐165 (Figure 1A), a SCLC cell line with very high constitutive expression of AVP 35. USF mutants did not stimulate proliferation in NCI‐H460, consistent with the lack of function associated with their loss of DNA binding or transcriptional activation domains 37 and their ability to reduce reporter gene activity dependent on the AVP promoter 23.…”

Section: Discussionsupporting

confidence: 82%

“…A pSG5 vector control (Stratagene) was used in transfections. 5 × 10 6 cells in 400 µl RPMI/10% BCS were transfected with 20 µg plasmid by electroporation, as previously described 35.…”

Section: Methodsmentioning

confidence: 99%

“…The AVP gene promoter contains several E‐box motifs 6–8 that demonstrate enhancer functions in response to USF‐1 and USF‐2 6, 23. The relative amount of USF that binds these E‐boxes in gel‐shift analysis was consistently greater for SCLC than NSCLC extracts 6 and corresponds with the levels of endogenous AVP expression 35 and AVP promoter‐dependent reporter gene expression in cell lines 6. Furthermore, transfection of USF‐2 into NSCLC increases USF‐2 abundance in whole cell extracts, binding to E‐boxes on gel‐shift analysis and AVP promoter‐driven reporter gene expression 23.…”

Section: Introductionmentioning

confidence: 99%

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Roles for USF‐2 in lung cancer proliferation and bronchial carcinogenesis

Ocejo-Garcia

Baokbah

Ashurst

et al. 2005

The Journal of Pathology

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The upstream stimulatory factors USF-1 and USF-2 dimerize to regulate transcription through E-box motifs in target genes. Although widely expressed, they can mediate tissue-specific transcription and we previously reported that USF-2 can enhance transcription of arginine vasopressin, a neuropeptide growth factor in small cell lung cancer. Here we determine the expression and role of USF-2 in lung cancer subtypes and examine USF-2 distribution in the bronchial epithelium. For a panel of 12 cell lines and 10 frozen human tumour samples, immunoblotting demonstrated that USF-2 expression was more frequent and abundant in small cell lung cancer than in non-small cell lung cancer. An immunohistochemical study of 108 formalin-fixed and paraffin-embedded human samples was undertaken to localize USF-2 expression and included 44 small cell and 32 non-small cell lung cancers, and 32 samples with bronchial dysplasia. USF-2 was restricted to ciliated cells in normal bronchial epithelium, but was more strongly expressed in dysplastic epithelium (72%) and certain lung cancer types, including small cell lung cancer (71%), squamous cell carcinoma (69%) and a large cell neuroendocrine carcinoma, but was less common in adenocarcinoma (11%). In a small series, expression was assessed adjacent to positively staining tumours; in phenotypically normal bronchial tissues, USF-2 was more highly expressed at 1 cm than at 5 cm from the tumour. Transient USF-2 overexpression in non-small cell lung cancer cell lines significantly stimulated in vitro cell proliferation; this response was most apparent for NCI-H460 (p < 0.005), reducing the mean cell doubling time from 19 to 16 h. Dominant-negative USF-2 mutants had no significant effect on cell growth. Taken together, these data suggest that USF-2 represents a relatively early molecular marker for the development of bronchial dysplasia and non-adenocarcinoma lung cancer. USF may also play a role in bronchial carcinogenesis, perhaps through promoting cell proliferation, although the genes through which this is regulated remain to be determined.

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Section: Discussionsupporting

confidence: 82%

“…A pSG5 vector control (Stratagene) was used in transfections. 5 × 10 6 cells in 400 µl RPMI/10% BCS were transfected with 20 µg plasmid by electroporation, as previously described 35.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Roles for USF‐2 in lung cancer proliferation and bronchial carcinogenesis

Ocejo-Garcia

Baokbah

Ashurst

et al. 2005

The Journal of Pathology

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“…The promoters for arginine vasopressin and the cell surface antigen CD24 genes have been suggested, but have not yet been tested with therapeutic genes. 8,9 The neuron-specific enolase promoter has been tried for gene therapy for SCLC, but expression of a suicide gene from this promoter could not mediate cell death. 10 Both tumor expression and serum levels of the gastrin releasing peptide (GRP) has been used as a marker for SCLC.…”

Section: Introductionmentioning

confidence: 99%

The insulinoma-associated 1: a novel promoter for targeted cancer gene therapy for small-cell lung cancer

Pedersen

Lan

et al. 2005

Cancer Gene Ther

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The insulinoma-associated 1 (INSM1) gene is expressed exclusively during early embryonal development, but has been found reexpressed at high levels in neuroendocrine tumors. The regulatory region of the INSM1 gene is therefore a potential candidate for regulating expression of a therapeutic gene in transcriptionally targeted cancer gene therapy against neuroendocrine tumors. We analyzed expression of a reporter gene from a 1.7 kb region of the INSM1 promoter in a large number of small-cell lung cancer (SCLC) cell lines. This INSM1 promoter region showed very high levels of expression in most of the SCLC cell lines and expression was absent in cell lines of non-neuroendocrine origin. Inclusion of the general transcriptional enhancer from SV40 compromised the specificity of the promoter and did not enhance transcription in most of the SCLC cell lines. For comparison, the region of the gastrin releasing peptide (GRP) previously suggested for SCLC gene therapy was analyzed in a similar manner. High expression was observed for a number of cell lines, but unlike for the INSM1 promoter, reporter gene expression from the GRP promoter did not correlate to the relative GRP mRNA levels, demonstrating that this region may not contain all necessary regulatory elements. Expression of the suicide gene herpes simplex virus thymidine kinase (HSV-TK) from the INSM1 promoter in combination with treatment with the prodrug ganciclovir (GCV) caused a significant increase in GCV sensitivity specifically in INSM1-expressing cell lines. The INSM1 promoter is therefore a potential novel tool for transcriptionally targeted gene therapy for neuroendocrine tumors.

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“…4 In support of this, other investigators have recently shown that vasopressin gene expression by this tumor type could result from the presence of an aberrant suppressor protein that normally binds to the upstream coding region for the vasopressin gene. 5 Part of our demonstration of vasopressin gene expression involved immunohistochemistry performed with polyclonal antibodies recognizing different regions of the provasopressin molecule. 6 Antibodies directed against vasopressin or the linkage regions between vasopressin and neurophysin each reacted positively with all SCLC tumors examined, while antibodies against the glycopeptide moiety (VAG) at the C-terminal region of provasopressin reacted positively with approximately 84% of tumors.…”

mentioning

confidence: 99%

Immunohistochemical Detection of NRSA on Small Cell Lung Cancer With a Monoclonal Antibody (MAG-1) That Recognizes the Carboxyl Terminus of Provasopressin

North¹,

Memoli²,

Keegan³

2005

Applied Immunohistochemistry &Amp; Molecular Morphology

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A single monoclonal antibody (MAG-1) directed against the C-terminal 18-amino acid region (VAGc18) of provasopressin was examined as an agent for recognizing the tumor-specific NRSA marker common to small cell lung cancer (SCLC) in formalin-fixed tissues with ABC immunohistochemistry. SCLC tumors were obtained from several tissue locations and included primary, metastatic, and recurrent disease. Positive staining was found in 91% of cases (53/58). All five of the unreactive tumors were of the lungs or chest wall, and there did not appear to be an association of this negativity with disease stage, age, or sex. Alternatively, almost all primary lesions, almost all metastatic lesions, and all recurrent lesions examined gave a positive reaction with MAG-1. For this study, vasopressin-producing cells of the human anterior hypothalamus served as a positive control, while negative controls comprised normal lung tissue, tumor that received MAG-1 in the presence of an excess of antigen (VAGc18 peptide), or tumor reacted with a commercial IgG1 isotype as primary antibody. All of the results indicate that MAG-1 can be effectively used to selectively identify the NRSA marker on almost all SCLC tumors, at all disease stages, and at all locations. Since all four tumors tested showing no reactivity with MAG-1 gave a positive reaction for synaptophysin, it is proposed that a combined use of MAG-1 with synaptophysin antibodies could allow all SCLC tumors to be detected by ABC immunohistochemistry.

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Tumour-specific arginine vasopressin promoter activation in small-cell lung cancer

Cited by 25 publications

References 29 publications

Roles for USF‐2 in lung cancer proliferation and bronchial carcinogenesis

Roles for USF‐2 in lung cancer proliferation and bronchial carcinogenesis

The insulinoma-associated 1: a novel promoter for targeted cancer gene therapy for small-cell lung cancer

Immunohistochemical Detection of NRSA on Small Cell Lung Cancer With a Monoclonal Antibody (MAG-1) That Recognizes the Carboxyl Terminus of Provasopressin

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