Chai, Yuan Lin
(2016)
Characterization Of Cal 27 Oral Squamous Carcinoma Cell Line As A Model For Cancer Stem Cell Study.
PhD thesis, Universiti Sains Malaysia.
Abstract
Peratusan sel stem kanser (CSC) agak rendah di kalangan populasi sel barah. Oleh itu, langkah-langkah pengasingan dan pengayaan CSC telah menjadi rutin bagi kerja-kerja penyelidikan yang melibatkan CSC. CAL 27 (ATCC® CRL-2095 TM) yang berasal daripada tisu lidah manusia, merupakan salah satu titisan sel epitelium skuamus karsinoma yang sering digunakan. Tujuan kajian ini dijalankan adalah untuk mewujudkan satu model titisan sel yang bersesuaian untuk mengkaji perkembangan CSC secara cekap, dengan mengurangkan masa bagi proses pengasingan dan juga langkah pengayaan sel. Kajian dijalankan untuk menentukan transformasi morfologi dan juga fisiologi sel titis CAL 27, terutamanya berkenaan dengan pengekspresian penanda sel stem mesenkima, tempoh masa penggandaan sel, rintangan terhadap rawatan, kebolehbalikan fenotaip, ciri invasif dan keupayaan metastatik. Selain itu, penglibatan lekatan sel dan peralihan epitelium-mesenkima (EMT), bersama dengan isyarat laluan Wnt juga diselidik. Hasilnya, pemanjangan sel CAL 27-O (sel CAL 27 dari generasi ke-6 dan seterusnya) dari sel CAL 27-Y (sel CAL 27 sebelum generasi ke-6) dikaitkan dengan peningkatan keupayaan invasif dan metastatik, rintangan rawatan yang lebih tinggi, pembalikan fenotaip yang mencirikan sel-sel epitelium, selain peningkatan kadar pembaharuan sel.
The percentage of cancer stem cells (CSCs) is relatively low in tumour bulk population. Therefore, isolation and enrichment steps for CSCs have become routine for research works involving CSC. CAL 27 (ATCC® CRL-2095 TM) is one of the commonly used squamous epithelial carcinoma cell lines derived from human tongue tissue. The present study was conducted with the objective to establish a suitable cell line model for studying CSC development efficiently, by reducing the time consuming isolation and enrichment steps. Experiments were carried out to determine the morphological as well as physiological transformations of CAL 27 cell line, particularly on the expression of mesenchymal stem cell markers, cell doubling time, treatment resistance, phenotype reversibility, invasiveness and metastatic ability. In addition to that, involvement of cell adhesion and epithelial-mesenchymal transition (EMT), together with Wnt signaling pathway were investigated. As for the results, elongation of CAL 27-O cells (CAL 27 cells from 6th passage onwards) from CAL 27-Y cells (CAL 27 cells below 6th passage) were found to be associated with enhanced invasive and metastatic capability, greater resistance to treatment, reversal of phenotype characterizing epithelial cells, besides enhanced proliferation.
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