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Frequent expression loss of Inter-alpha-trypsin inhibitor heavy chain (ITIH) genes in multiple human solid tumors: a systematic expression analysis.

Hamm A, Veeck J, Bektas N, Wild PJ, Hartmann A, Heindrichs U, Kristiansen G, Werbowetski-Ogilvie T, Del Maestro R, Knuechel R, Dahl E - BMC Cancer (2008)

Bottom Line: We found that ITIH genes are clearly downregulated in multiple human solid tumors, including breast, colon and lung cancer.Thus, ITIH genes may represent a family of putative tumor suppressor genes that should be analyzed in greater detail in the future.We found a strong correlation (p < 0.001) between ITIH2 expression and estrogen receptor (ER) expression indicating that ER may be involved in the regulation of this ECM molecule.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Pathology, University Hospital of RWTH Aachen, Aachen, Germany. alexander.hamm@rwth-aachen.de

ABSTRACT

Background: The inter-alpha-trypsin inhibitors (ITI) are a family of plasma protease inhibitors, assembled from a light chain - bikunin, encoded by AMBP - and five homologous heavy chains (encoded by ITIH1, ITIH2, ITIH3, ITIH4, and ITIH5), contributing to extracellular matrix stability by covalent linkage to hyaluronan. So far, ITIH molecules have been shown to play a particularly important role in inflammation and carcinogenesis.

Methods: We systematically investigated differential gene expression of the ITIH gene family, as well as AMBP and the interacting partner TNFAIP6 in 13 different human tumor entities (of breast, endometrium, ovary, cervix, stomach, small intestine, colon, rectum, lung, thyroid, prostate, kidney, and pancreas) using cDNA dot blot analysis (Cancer Profiling Array, CPA), semiquantitative RT-PCR and immunohistochemistry.

Results: We found that ITIH genes are clearly downregulated in multiple human solid tumors, including breast, colon and lung cancer. Thus, ITIH genes may represent a family of putative tumor suppressor genes that should be analyzed in greater detail in the future. For an initial detailed analysis we chose ITIH2 expression in human breast cancer. Loss of ITIH2 expression in 70% of cases (n = 50, CPA) could be confirmed by real-time PCR in an additional set of breast cancers (n = 36). Next we studied ITIH2 expression on the protein level by analyzing a comprehensive tissue micro array including 185 invasive breast cancer specimens. We found a strong correlation (p < 0.001) between ITIH2 expression and estrogen receptor (ER) expression indicating that ER may be involved in the regulation of this ECM molecule.

Conclusion: Altogether, this is the first systematic analysis on the differential expression of ITIH genes in human cancer, showing frequent downregulation that may be associated with initiation and/or progression of these malignancies.

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Related in: MedlinePlus

Differential expression of the ITIH gene family in 13 different human tumor entities. Gene probes for ITIH genes, as well as the AMBP and TNFAIP6 genes, were hybridized to a Cancer Profiling Array (Clontech, Heidelberg, Germany) containing spotted cDNAs from 241 human tumors and 241 corresponding human normal tissues. See text for details and Table 4 for a detailed densitometric evaluation of the hybridization signals.
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Figure 2: Differential expression of the ITIH gene family in 13 different human tumor entities. Gene probes for ITIH genes, as well as the AMBP and TNFAIP6 genes, were hybridized to a Cancer Profiling Array (Clontech, Heidelberg, Germany) containing spotted cDNAs from 241 human tumors and 241 corresponding human normal tissues. See text for details and Table 4 for a detailed densitometric evaluation of the hybridization signals.

Mentions: Next, these gene probes were used for dot blot hybridization on a Cancer Profiling Array (CPA). The CPA contains spotted cDNAs from 241 tumor and 241 matched normal tissue samples representing 13 different human tumor entities. These dot blot arrays are highly sensitive in the detection of even rare transcripts since a considerable amount of cDNA (10–50 μg) has been spotted in a very small area (1 mm in diameter). In analysis of differential expression, the percentage of up-regulation or down-regulation was defined according to the well-established fold change two approach (FC2, see Material and Methods). Interestingly, each member of the ITIH gene family except ITIH1 (data not shown) exhibited highly differential expression in a remarkable number of human tumor entities (Figure 2). These differential expression patterns nearly exclusively presented as loss of ITIH mRNAs in tumor tissues. An overview of the complete expression data is provided in Table 4. In contrast to the highly differential expression of ITIH genes, AMBP and TNFAIP6 exhibited a much broader expression pattern with differential gene expression restricted to kidney tissue. In detail, the expression patterns of the analyzed genes on the CPA presented as follows:


Frequent expression loss of Inter-alpha-trypsin inhibitor heavy chain (ITIH) genes in multiple human solid tumors: a systematic expression analysis.

Hamm A, Veeck J, Bektas N, Wild PJ, Hartmann A, Heindrichs U, Kristiansen G, Werbowetski-Ogilvie T, Del Maestro R, Knuechel R, Dahl E - BMC Cancer (2008)

Differential expression of the ITIH gene family in 13 different human tumor entities. Gene probes for ITIH genes, as well as the AMBP and TNFAIP6 genes, were hybridized to a Cancer Profiling Array (Clontech, Heidelberg, Germany) containing spotted cDNAs from 241 human tumors and 241 corresponding human normal tissues. See text for details and Table 4 for a detailed densitometric evaluation of the hybridization signals.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2268946&req=5

Figure 2: Differential expression of the ITIH gene family in 13 different human tumor entities. Gene probes for ITIH genes, as well as the AMBP and TNFAIP6 genes, were hybridized to a Cancer Profiling Array (Clontech, Heidelberg, Germany) containing spotted cDNAs from 241 human tumors and 241 corresponding human normal tissues. See text for details and Table 4 for a detailed densitometric evaluation of the hybridization signals.
Mentions: Next, these gene probes were used for dot blot hybridization on a Cancer Profiling Array (CPA). The CPA contains spotted cDNAs from 241 tumor and 241 matched normal tissue samples representing 13 different human tumor entities. These dot blot arrays are highly sensitive in the detection of even rare transcripts since a considerable amount of cDNA (10–50 μg) has been spotted in a very small area (1 mm in diameter). In analysis of differential expression, the percentage of up-regulation or down-regulation was defined according to the well-established fold change two approach (FC2, see Material and Methods). Interestingly, each member of the ITIH gene family except ITIH1 (data not shown) exhibited highly differential expression in a remarkable number of human tumor entities (Figure 2). These differential expression patterns nearly exclusively presented as loss of ITIH mRNAs in tumor tissues. An overview of the complete expression data is provided in Table 4. In contrast to the highly differential expression of ITIH genes, AMBP and TNFAIP6 exhibited a much broader expression pattern with differential gene expression restricted to kidney tissue. In detail, the expression patterns of the analyzed genes on the CPA presented as follows:

Bottom Line: We found that ITIH genes are clearly downregulated in multiple human solid tumors, including breast, colon and lung cancer.Thus, ITIH genes may represent a family of putative tumor suppressor genes that should be analyzed in greater detail in the future.We found a strong correlation (p < 0.001) between ITIH2 expression and estrogen receptor (ER) expression indicating that ER may be involved in the regulation of this ECM molecule.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Pathology, University Hospital of RWTH Aachen, Aachen, Germany. alexander.hamm@rwth-aachen.de

ABSTRACT

Background: The inter-alpha-trypsin inhibitors (ITI) are a family of plasma protease inhibitors, assembled from a light chain - bikunin, encoded by AMBP - and five homologous heavy chains (encoded by ITIH1, ITIH2, ITIH3, ITIH4, and ITIH5), contributing to extracellular matrix stability by covalent linkage to hyaluronan. So far, ITIH molecules have been shown to play a particularly important role in inflammation and carcinogenesis.

Methods: We systematically investigated differential gene expression of the ITIH gene family, as well as AMBP and the interacting partner TNFAIP6 in 13 different human tumor entities (of breast, endometrium, ovary, cervix, stomach, small intestine, colon, rectum, lung, thyroid, prostate, kidney, and pancreas) using cDNA dot blot analysis (Cancer Profiling Array, CPA), semiquantitative RT-PCR and immunohistochemistry.

Results: We found that ITIH genes are clearly downregulated in multiple human solid tumors, including breast, colon and lung cancer. Thus, ITIH genes may represent a family of putative tumor suppressor genes that should be analyzed in greater detail in the future. For an initial detailed analysis we chose ITIH2 expression in human breast cancer. Loss of ITIH2 expression in 70% of cases (n = 50, CPA) could be confirmed by real-time PCR in an additional set of breast cancers (n = 36). Next we studied ITIH2 expression on the protein level by analyzing a comprehensive tissue micro array including 185 invasive breast cancer specimens. We found a strong correlation (p < 0.001) between ITIH2 expression and estrogen receptor (ER) expression indicating that ER may be involved in the regulation of this ECM molecule.

Conclusion: Altogether, this is the first systematic analysis on the differential expression of ITIH genes in human cancer, showing frequent downregulation that may be associated with initiation and/or progression of these malignancies.

Show MeSH
Related in: MedlinePlus