Int J Immunopathol Pharmacol.2013 Jan-Mar;26(1):223-228

Expression of ADAM28 and iGFBP-3 genes in patients with colorectal cancer – a preliminary report

Nowakowska-Zajdel E1, Mazurek U2, Wierzgoń J3, Kokot T1, Fatyga E1, Ziółko E1, Klakla K1, Błażelonis A1, Waniczek D4, Głogowski Ł5, Kozowicz A1, Niedworok E6, Muc-Wierzgoń M1

1Department of Internal Medicine   Medical University of Silesia, Katowice, Poland

2Department of Molecular Biology Medical University of Silesia, Katowice, Poland

3Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Department of Oncologic and Reconstructive Surgery Gliwice, Poland

4Department of Surgery, Medical University of Silesia, Katowice, Poland

5Department of Clinical Oncology, Regional Hospital No.4, Bytom, Poland

6Department of Human Nutrition, Medical University of Silesia, Katowice, Poland

 

Abstract:

Adamalisynes (ADAMs) play an important role in inter-membrane interactions, cell adhesion and fusion processes and protein shedding from the cell surface. Many reports indicate that members of the ADAMs family are overexpressed in human cancer. The aim of the present study was to evaluate ADAM28 and Insulin Like Growth Factor Binding Protein-3 (IGFBP-3) gene expression in colorectal carcinoma tissues with regard to the overweight or obese status of the patients using an oligonucleotide microarray technique. Fresh tissue specimens were obtained from colorectal cancer (CRC) patients during surgical treatment. 18 specimens from tumour and 18 normal tissue specimens from colorectal cancer patients at clinical stages III and IV were analysed. The examined patients were divided into two groups; those with BMI≥25 and those with normal BMI. The control group consisted of 18 specimens of non-neoplastic colon tissues, which were divided between overweight/obese and normal body weight patients. The gene transcriptional activity from the specimens was analysed using an oligonucleotide microarray technique. Microarrays and rinsing and marking solutions were prepared according to the procedure in the Gene Expression Analysis Technical Manual. Conclusions: 1. Change of ADAM28 and IGFBP-3 genes expression are present  in the normal tissue in overweight/obese patients with colorectal cancer only. 2. The observed molecular variability of ADAM28 and IGFBP-3 expression may be an initial process of cancer proliferation. 3. The histopathologically normal surgical margin in this group of patients was not equal to the molecular margin.

Key words: adamalisynes, IGF system, colorectal cancer, surgical and molecular margin

PMID: 23527725

 

Supplement

After activation of the precursor with matrix metalloproteinase-7 (MMP-7), ADAM28 releases insulin-like growth factor-1 (IGF1) from the IGF1/IGFBP-3 complex by selective digestion of IGFBP-3 into two major fragments. IGF1 induces phosphorylation of IGF1R and promotes cell proliferation through the ERK pathway [1]. Some research has indicated that high circulating IGF1 levels and an increased IGF1/IGFBP3 ratio disturbs GH/IGF1 homeostasis, which could be an indicator of risk for cancer development [2]. IGFBP-3 binds and sequesters the majority of the IGF1 ligands. It has been shown to inhibit proliferation and induce apoptosis in human colon cancer cells in vitro and in an experimental CRC animal model [3]. Epidemiological studies showed that higher circulating IGF1 levels and lower IGFBP-3 levels independently correlate with increased CRC risk. IGFBP-3 suppresses tumour growth and angiogenesis by both IGF-dependent and IGF-independent mechanisms [4].

In our study eighteen specimens from tumour and  eighteen specimens from normal tissue of CRC patients (adenocarcinoma) at clinical stage III and IV were analysed. The examined patients were divided into two groups: nine patients with BMI≥25 (labelled O-WSZ) and nine with normal BMI (labelled WSZ). The control group consisted of 18 specimens of non-neoplastic colon tissues taken from the surgical margin of the tumour that were also divided according to overweight/obese (labelled O-K) and normal body weight patients (labelled K).

The results of molecular (according to the procedure of the Gene Expression Analysis Technical Manual) and statistical analysis (Gene Spring software for the microarray) was indicated that the comparison of transcriptomes from the specimens of normal tissue and colorectal cancer (O-K vs O-WSZ) was performed to determine significant differences in gene transcription. The expression level is mapped by the colour intensity (Fig1). The fluorescence intensity map indicates the differentiated genes in the IGF system. Change of ADAM28 and IGFBP-3 genes expression were observed only  in the normal tissue in overweight/obese patients with colorectal cancer.

1Fig.1. Intensity map of fluorescence signals of O-K vs O-WSZ differentiating genes (blue color – gene silencing, red color – gene overexpression)

In the normal tissue of colorectal cancer patients with overweight/obese, the change of ADAM28 and IGFBP-3 gene expressions  (perhaps overexpression and silencing, respectively) was observed, which led to an increase in IGF1 bioactivity in the microenvironment of the tissue. The above-mentioned changes were observed in the histopathologically normal specimen (surgical margin) but not the carcinoma tissue. Notably, the expression of these genes in tumour specimens independently of BMI and normal tissue of the patients with BMI<25 were indistinguishable. In our study, normal expression of ADAM28 and IGFBP-3 genes in tumours may indicate the presence of reverse processes (necrosis, heterogeneity in the study sample, microenvironmental lesions in the tumour blood circulation, etc.) with no proliferative activity, whereas the altered activity in the genes in the surrounding tissues of the tumour (microscopically normal surgical margin) may suggest proliferation of cancerous cells at the site. Confirmation of gene expression using the PCR method and  increasing the number of patients in the study are necessary.

 

References

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