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Clonality analysis of synchronous lesions of cervical carcinoma based on X chromosome inactivation polymorphism, human papillomavirus type 16 genome mutations, and loss of heterozygosity.

Hu X, Pang T, Asplund A, Pontén J, Nistér M - J. Exp. Med. (2002)

Bottom Line: Microdissection was performed on 24 samples from this case, representing the entire lesional situation.The combination of different X chromosome inactivation patterns, two HPV16 point mutations, and LOH at three genomic microsatellite loci, led to the identification of five different "monoclonal" lesions (CIN II, CIN III, and invasive carcinoma nests) and five different "polyclonal" areas (CIN II and normal squamous epithelium).Our results also supported the view that HPV16 as a "field factor" causes cervical carcinoma, which is probably promoted by the loss of chromosomal material as indicated by the LOH.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-751 85, Uppsala, Sweden. hu.xinrong@genpat.uu.se

ABSTRACT
One of the most common forms of carcinoma in women, cervical invasive squamous cell carcinoma (CIC), often coexists with multiple lesions of cervical intraepithelial neoplasia (CIN). CIC and CIN show heterogeneity with respect to both histopathology and biology. To understand the causes, origin, and model of progression of cervical carcinoma, we assessed the clonality of a case with multiple synchronous lesions by analyzing X chromosome inactivation polymorphism, human papillomavirus type 16 (HPV16) sequence variation/mutations, and loss of heterozygosity (LOH). Microdissection was performed on 24 samples from this case, representing the entire lesional situation. The combination of different X chromosome inactivation patterns, two HPV16 point mutations, and LOH at three genomic microsatellite loci, led to the identification of five different "monoclonal" lesions (CIN II, CIN III, and invasive carcinoma nests) and five different "polyclonal" areas (CIN II and normal squamous epithelium). This finding indicated that CIC can originate from multiple precursor cells, from which some clones might progress via multiple steps, namely via CIN II and CIN III, whereas others might develop independently and possibly directly from the carcinoma precursor cells. Our results also supported the view that HPV16 as a "field factor" causes cervical carcinoma, which is probably promoted by the loss of chromosomal material as indicated by the LOH.

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Related in: MedlinePlus

Topography and histopathology of microdissected samples. Sides with F, D, or B (names of tissue block) are external ostium portions and the opposite sides are facing endocervix. #, H2−; arrows point to the foci where samples (named H2-1, -2, etc.) were microdissected; N, normal squamous epithelium; G, gland epithelium; S, stroma; IC, invasive carcinoma; Sup. Inv., superficially invasive carcinoma.
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fig1: Topography and histopathology of microdissected samples. Sides with F, D, or B (names of tissue block) are external ostium portions and the opposite sides are facing endocervix. #, H2−; arrows point to the foci where samples (named H2-1, -2, etc.) were microdissected; N, normal squamous epithelium; G, gland epithelium; S, stroma; IC, invasive carcinoma; Sup. Inv., superficially invasive carcinoma.

Mentions: 6 μm of serial cryosections were prepared from parts B, D, and F, and stained briefly with Mayer's hematoxylin. Multiple microdissections were performed on invasive cancer nests CIN II and CIN III, normal epithelium, and glands and stroma from different areas in a representative section for each tissue block. Altogether 24 samples (H2-1–24) were taken covering the whole lesional area. When it was necessary to repeat the microdissection procedure it was done using immediately adjacent sections that represented the same areas as originally chosen for sampling. Sample number, location, and morphology are summarized in Fig. 1 . All invasive lesions were distinctly demarcated from stroma and CINs from adjacent normal epithelium (Fig. 2) . Admixture of normal epithelial, stromal, or inflammatory cells was insignificant judging by careful examination under the microscope. Microdissection was performed with a scalpel, and the blade was changed after each microdissection. The microdissected pieces were transferred to Eppendorf tubes containing 50 μl of PCR buffer II (PerkinElmer; Roche Molecular System). Each sample contained ∼500–1,000 cells (22).


Clonality analysis of synchronous lesions of cervical carcinoma based on X chromosome inactivation polymorphism, human papillomavirus type 16 genome mutations, and loss of heterozygosity.

Hu X, Pang T, Asplund A, Pontén J, Nistér M - J. Exp. Med. (2002)

Topography and histopathology of microdissected samples. Sides with F, D, or B (names of tissue block) are external ostium portions and the opposite sides are facing endocervix. #, H2−; arrows point to the foci where samples (named H2-1, -2, etc.) were microdissected; N, normal squamous epithelium; G, gland epithelium; S, stroma; IC, invasive carcinoma; Sup. Inv., superficially invasive carcinoma.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193730&req=5

fig1: Topography and histopathology of microdissected samples. Sides with F, D, or B (names of tissue block) are external ostium portions and the opposite sides are facing endocervix. #, H2−; arrows point to the foci where samples (named H2-1, -2, etc.) were microdissected; N, normal squamous epithelium; G, gland epithelium; S, stroma; IC, invasive carcinoma; Sup. Inv., superficially invasive carcinoma.
Mentions: 6 μm of serial cryosections were prepared from parts B, D, and F, and stained briefly with Mayer's hematoxylin. Multiple microdissections were performed on invasive cancer nests CIN II and CIN III, normal epithelium, and glands and stroma from different areas in a representative section for each tissue block. Altogether 24 samples (H2-1–24) were taken covering the whole lesional area. When it was necessary to repeat the microdissection procedure it was done using immediately adjacent sections that represented the same areas as originally chosen for sampling. Sample number, location, and morphology are summarized in Fig. 1 . All invasive lesions were distinctly demarcated from stroma and CINs from adjacent normal epithelium (Fig. 2) . Admixture of normal epithelial, stromal, or inflammatory cells was insignificant judging by careful examination under the microscope. Microdissection was performed with a scalpel, and the blade was changed after each microdissection. The microdissected pieces were transferred to Eppendorf tubes containing 50 μl of PCR buffer II (PerkinElmer; Roche Molecular System). Each sample contained ∼500–1,000 cells (22).

Bottom Line: Microdissection was performed on 24 samples from this case, representing the entire lesional situation.The combination of different X chromosome inactivation patterns, two HPV16 point mutations, and LOH at three genomic microsatellite loci, led to the identification of five different "monoclonal" lesions (CIN II, CIN III, and invasive carcinoma nests) and five different "polyclonal" areas (CIN II and normal squamous epithelium).Our results also supported the view that HPV16 as a "field factor" causes cervical carcinoma, which is probably promoted by the loss of chromosomal material as indicated by the LOH.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-751 85, Uppsala, Sweden. hu.xinrong@genpat.uu.se

ABSTRACT
One of the most common forms of carcinoma in women, cervical invasive squamous cell carcinoma (CIC), often coexists with multiple lesions of cervical intraepithelial neoplasia (CIN). CIC and CIN show heterogeneity with respect to both histopathology and biology. To understand the causes, origin, and model of progression of cervical carcinoma, we assessed the clonality of a case with multiple synchronous lesions by analyzing X chromosome inactivation polymorphism, human papillomavirus type 16 (HPV16) sequence variation/mutations, and loss of heterozygosity (LOH). Microdissection was performed on 24 samples from this case, representing the entire lesional situation. The combination of different X chromosome inactivation patterns, two HPV16 point mutations, and LOH at three genomic microsatellite loci, led to the identification of five different "monoclonal" lesions (CIN II, CIN III, and invasive carcinoma nests) and five different "polyclonal" areas (CIN II and normal squamous epithelium). This finding indicated that CIC can originate from multiple precursor cells, from which some clones might progress via multiple steps, namely via CIN II and CIN III, whereas others might develop independently and possibly directly from the carcinoma precursor cells. Our results also supported the view that HPV16 as a "field factor" causes cervical carcinoma, which is probably promoted by the loss of chromosomal material as indicated by the LOH.

Show MeSH
Related in: MedlinePlus