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Sains Malaysiana 52(5)(2023):
http://doi.org/10.17576/jsm-2023-5205-12
Proportion of CD44+ Subset of Tumour Cells in Single Cell Suspension
Prepared from FFPET Sections Directly Correlates with Histological Subtyping of
Head and Neck Squamous Cell Carcinoma
(Bahagian CD44+ Subset Sel Tumor
dalam Suspensi Sel Tunggal Disediakan daripada Bahagian FFPET Berkorelasi Terus
dengan Pengetipan Histologi Karsinoma Sel Skuamosa Kepala dan Leher)
MUHAMMAD
KASHIF1,2,*, SADIA MINHAS3, SHAH JAHAN1, AFRA
SAMAD4, FAHEEM SHAHZAD1, ROMEEZA TAHIR1, ABDUL
HANAN NAGI5 & NADEEM AFZAL1
1Department
of Immunology, University of Health Sciences, 54000, Khyaban e Jamia Punjab, Lahore, Pakistan
2Department
of Oral Pathology, Bakhtawar Amin Medical and Dental
College, Northern Bypass Road, 60000, Multan, Pakistan
3Department
of Oral Pathology, Akhtar Saeed Medical and Dental College, Bahria Town, 54000, Lahore, Pakistan
4Department
of Histopathology, Multan Medical and Dental College, 60000, Southern bypass
road, Multan, Pakistan
5Chughtai Laboratory, Lahore, Pakistan
Received: 19 December 2022/Accepted: 17 April
2023
Abstract
CD44 expression in tumours imparts
potential to progress, metastasize, recurrence, and resistance against
antineoplastic therapy. In this study, we sought to describe the variation in
the immuno-expression and numeration of MDR1+ and CD44+ potential cancer stem cells in different histological grades and subtypes of
head and neck squamous cell carcinoma (HNSCC). Flow-cytometric analysis was
performed on single cell suspension prepared from formalin fixed paraffin
embedded tissue (FFPET) sections of HNSCC using anti-CD44 and anti-MDR1/ABCB-1
primary monoclonal antibodies. Immunohistochemical (IHC) staining was also carried out using both of these antibodies on HNSCC
tissue sections mounted on super frosted glass slides. On immunohistochemical analysis, the
mean IRS for CD44 and MDR1 were 8.6364 ±3.02114 and 1.5909 ±1.27674
respectively. When mean immune-expression scores of CD44 antibody and
MDR1/ABCB-1 were compared with histological grades and subtypes of HNSCC, the
relationship was found to be statistically insignificant. Interestingly, a
strong statistical difference (p =
0.000) was observed when the mean score of subset of dysplastic squamous epithelial cells
with characteristics of cell stemness (CD326+CD44+)
was compared among different histological subtypes of HNSCC using flowcytometric analysis. While no statistically significant
association was observed when the mean score for subset of dysplastic cells
with potential of drug resistance (CD44+MDR1+) was
compared among different histological subtypes of HNSCC. Although potential cancer stem cell marker CD44 and the
multidrug resistance maker ABCB-1/MDR1 co-expressed in HNSCC but the proportion
of CD326+CD44+ subset of tumour cells (potential cancer stem cells/CSCs) significantly correlates with least
aggressive to more aggressive tumour subtypes.
Keywords: CD44; CSCs; FFPET; flowcytometry;
HNSCC; immunohistochemistry; multidrug resistance; OSCC
Abstract
Pengekspresan CD44 dalam tumor
memberikannya potensi untuk berkembang, bermetastasis, berulang dan berintangan
terhadap terapi antineoplastik. Dalam kajian ini, kami ingin menerangkan variasi dalam
pengekspresan imun dan penbilangan sel stem kanser berpotensi MDR1+ dan CD44+ dalam gred histologi yang berbeza dan pengetipan karsinoma
sel skuamosa kepala dan leher (HNSCC). Analisis aliran-sitometri dilakukan pada penggantungan
sel tunggal yang disediakan daripada bahagian tisu terbenam parafin tetap
formalin (FFPET) HNSCC menggunakan antibodi monoklonal utama anti-CD44 dan
anti-MDR1/ABCB-1. Pewarnaan imunohistokimia (IHC) juga dilakukan
menggunakan kedua-dua antibodi ini pada bahagian tisu HNSCC yang dipasang pada
slaid kaca super beku. Pada analisis imunohistokimia, purata IRS untuk CD44 dan
MDR1 masing-masing ialah 8.6364 ±3.02114 dan 1.5909 ±1.27674. Apabila min
skor pengekspresan imun antibodi CD44 dan MDR1/ABCB-1 dibandingkan dengan gred
histologi dan pengetipan HNSCC, hubungan itu didapati tidak signifikan secara
statistik. Menariknya, perbezaan statistik yang kuat (p = 0.000)
diperhatikan apabila skor min subset sel epitelium skuamosa displastik dengan
ciri-ciri pensteman sel (CD326+CD44+) dibandingkan antara
pengetipan histologi HNSCC yang berbeza menggunakan analisis sitometri aliran. Walaupun tiada
perkaitan yang signifikan secara statistik, diperhatikan apabila skor min bagi
subset sel displastik dengan potensi rintangan drug (CD44+MDR1+)
dibandingkan antara pengetipan histologi HNSCC yang berbeza. Walaupun
penanda sel stem kanser berpotensi CD44 dan pembuat rintangan pelbagai drug
ABCB-1/MDR1 dinyatakan bersama dalam HNSCC tetapi perkadaran subset CD326+CD44+ sel tumor (sel stem berpotensi kanser/CSC) berkorelasi dengan ketara dengan
tumor yang paling tidak agresif kepada pengetipan tumor yang lebih agresif.
Kata kunci: CD44;
CSCs; FFPET; HNSCC; imunohistokimia; OSCC; rintangan pelbagai ubat; sitometri
aliran
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*Corresponding author; email: drkashifazam@gmail.com
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