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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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ITIH2 immunohistochemistry on TMA derived from normal and cancerous breast tissue. A+B: Strong cytoplasmic staining is seen in normal epithelial cells of the mammary gland. C+D: Ductal carcinoma in-situ (high grade type) with moderate focal cytoplasmic staining and normal, partially hyperplastic gland epithelium with strong cytoplasmic staining (see arrows). E-H: Invasive ductal carcinoma with either negative (E, F) or strong ITIH2 staining (G, H). Magnification: 100× (A, C, E, G), 400× (B, D, F, H).
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Figure 4: ITIH2 immunohistochemistry on TMA derived from normal and cancerous breast tissue. A+B: Strong cytoplasmic staining is seen in normal epithelial cells of the mammary gland. C+D: Ductal carcinoma in-situ (high grade type) with moderate focal cytoplasmic staining and normal, partially hyperplastic gland epithelium with strong cytoplasmic staining (see arrows). E-H: Invasive ductal carcinoma with either negative (E, F) or strong ITIH2 staining (G, H). Magnification: 100× (A, C, E, G), 400× (B, D, F, H).

Mentions: Immunohistochemical analysis was applied to compare ITIH2 protein expression in normal and malignant breast tissue. A tissue microarray containing 185 invasive breast carcinomas, 2 carcinomas in-situ (DCIS), and 28 normal breast tissue samples was used. Intensity and quantity of immunohistochemical staining was evaluated using a semiquantitative immunoreactivity score (IRS) [47]. ITIH2 expression was clearly detectable in the epithelium of normal breast tissue (Figure 4A and 4B). ITIH2 expression was maintained in hyperplastic gland epithelium and ductal carcinoma in-situ (DCIS), however, ITIH2 expression was somewhat weaker in DCIS than in normal tissue (Figure 4C and 4D). In 44% (81/185) of invasive carcinomas of the breast, ITIH2 expression was strongly reduced or completely lost (Figure 4E and 4F) while 56% of invasive carcinomas (104/185) maintained moderate to strong ITIH2 expression (Figure 4G and 4H).


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)

ITIH2 immunohistochemistry on TMA derived from normal and cancerous breast tissue. A+B: Strong cytoplasmic staining is seen in normal epithelial cells of the mammary gland. C+D: Ductal carcinoma in-situ (high grade type) with moderate focal cytoplasmic staining and normal, partially hyperplastic gland epithelium with strong cytoplasmic staining (see arrows). E-H: Invasive ductal carcinoma with either negative (E, F) or strong ITIH2 staining (G, H). Magnification: 100× (A, C, E, G), 400× (B, D, F, H).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: ITIH2 immunohistochemistry on TMA derived from normal and cancerous breast tissue. A+B: Strong cytoplasmic staining is seen in normal epithelial cells of the mammary gland. C+D: Ductal carcinoma in-situ (high grade type) with moderate focal cytoplasmic staining and normal, partially hyperplastic gland epithelium with strong cytoplasmic staining (see arrows). E-H: Invasive ductal carcinoma with either negative (E, F) or strong ITIH2 staining (G, H). Magnification: 100× (A, C, E, G), 400× (B, D, F, H).
Mentions: Immunohistochemical analysis was applied to compare ITIH2 protein expression in normal and malignant breast tissue. A tissue microarray containing 185 invasive breast carcinomas, 2 carcinomas in-situ (DCIS), and 28 normal breast tissue samples was used. Intensity and quantity of immunohistochemical staining was evaluated using a semiquantitative immunoreactivity score (IRS) [47]. ITIH2 expression was clearly detectable in the epithelium of normal breast tissue (Figure 4A and 4B). ITIH2 expression was maintained in hyperplastic gland epithelium and ductal carcinoma in-situ (DCIS), however, ITIH2 expression was somewhat weaker in DCIS than in normal tissue (Figure 4C and 4D). In 44% (81/185) of invasive carcinomas of the breast, ITIH2 expression was strongly reduced or completely lost (Figure 4E and 4F) while 56% of invasive carcinomas (104/185) maintained moderate to strong ITIH2 expression (Figure 4G and 4H).

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