J Pathol Transl Med.  2018 Sep;52(5):314-322. 10.4132/jptm.2018.07.17.

An Immunohistochemical and Polarizing Microscopic Study of the Tumor Microenvironment in Varying Grades of Oral Squamous Cell Carcinoma

• Khalid A
• Siddiqui S
• Bordoloi B
• Faizi N
• Samadi F
• Saeed N

• KMID: 2422098
• DOI: http://doi.org/10.4132/jptm.2018.07.17

Abstract

BACKGROUND
Invasion of epithelial cells into the connective tissue brings about massive morphological and architectural changes in the underlying stroma. Myofibroblasts reorganize the stroma to facilitate the movement of tumor cells leading to metastasis. The aim of this study was to determine the number and pattern of distribution of myofibroblasts and the qualitative and quantitative change that they cause in the collagen present in the stroma in various grades of oral squamous cell carcinoma (OSCC).
METHODS
The study was divided into two groups with group I (test group, 65 cases) consisting of 29 cases of well-differentiated squamous cell carcinoma, 25 moderately differentiated SCC, and 11 poorly differentiated SCC, and group II (control group) consisting of 11 cases of normal mucosa. Sections from each sample were stained with anti-α-smooth muscle actin (α-SMA) antibodies, hematoxylin and eosin, and Picrosirius red. Several additional sections from each grade of OSCC were stained with Masson's trichrome to observe the changes in collagen. For the statistical analysis, Fisher's exact test, Tukey's post hoc honest significant difference test, ANOVA, and the chi-square test were used, and p < .05 was considered statistically significant.
RESULTS
As the tumor stage progressed, an increase in the intensity α-SMA expression was seen, and the network pattern dominated in more dedifferentiated carcinomas. The collagen fibers became thin, loosely packed, and haphazardly aligned with progressing cancer. Additionally, the mean area fraction decreased, and the fibers attained a greenish yellow hue and a weak birefringence when observed using polarizing light microscopy.
CONCLUSIONS
Myofibroblasts bring about numerous changes in collagen. As cancer progresses, there isincrease in pathological collagen,which enhances the movement of cells within the stroma.

Keyword

Myofibroblast; Collagen; Oral squamous cell carcinoma; α-Smooth muscle actin; Picrosirius red; Polarizing microscope

MeSH Terms

Actins
Antibodies
Birefringence
Carcinoma, Squamous Cell*
Collagen
Connective Tissue
Eosine Yellowish-(YS)
Epithelial Cells*
Hematoxylin
Microscopy
Mucous Membrane
Myofibroblasts
Neoplasm Metastasis
Tumor Microenvironment*
Actins
Antibodies
Collagen
Eosine Yellowish-(YS)
Hematoxylin

Figure

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  Fig. 1. Light microscopy image of well-differentiated squamous cell carcinoma showing mild α-smooth muscle actin positivity in the stroma. Myofibroblasts can be seen distributed in a focal pattern.

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  Fig. 2. Light microscopy image of moderately differentiated squamous cell carcinoma showing intense α-smooth muscle actin positivity in the stroma. Myofibroblasts can be seen distributed in a spindle pattern.

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  Fig. 3. Light microscopy image of poorly differentiated squamous cell carcinoma showing intense α-smooth muscle actin positivity in the stroma. Myofibroblasts can be seen distributed in a network pattern.

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  Fig. 4. Light microscopy image of a Masson’s trichrome–stained section of well-differentiated oral squamous cell carcinoma showing densely packed collagen fibersexhibiting a parallel arrangement.

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  Fig. 5. Light microscopy image of a Masson’s trichrome–stained section of moderately differentiated oral squamous cell carcinoma showing haphazardly arranged collagen fibers.

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  Fig. 6. Light microscopy image of a Masson’s trichrome–stained section of poorly differentiated oral squamous cell carcinoma showing haphazardly arranged, loosely packed collagen fibers.

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  Fig. 7. Polarizing light microscopy image of a Picrosirius redstained section of well-differentiated oral squamous cell carcinoma showing densely packed collagen fibers (arrow) exhibiting a parallel arrangement and orange-red birefringence.

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  Fig. 8. Polarizing light microscopy image of a Picrosirius redstained section of moderately differentiated oral squamous cell carcinoma showing haphazardly arranged, loosely packed collagen fibers (arrow) exhibiting yellowish-orange birefringence.

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  Fig. 9. Polarizing light microscopy image of a Picrosirius redstained section of poorly differentiated oral squamous cell carcinoma showing haphazardly arranged, loosely packed collagen fibers (arrow) exhibiting greenish-yellow birefringence.

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