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ORIGINAL ARTICLE
Year : 2021  |  Volume : 12  |  Issue : 3  |  Page : 247-254

Cancer stem cell traits in tumor spheres derived from primary laryngeal carcinoma cell lines


1 Central Research Laboratory, Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences and Research Centre; Department of Biotechnology, KLE Technological University, BVB Campus, Hubballi, India
2 Department of Biotechnology, KLE Technological University, BVB Campus, Hubballi, India
3 Central Research Laboratory, Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences and Research Centre, Hubballi, India
4 Department of Oral Pathology, Maratha Mandal's N. G. Halgekar Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India
5 Central Research Laboratory, Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences and Research Centre, Hubballi; Department of Pharmaceutics, Maratha Mandal's College of Pharmacy, Belagavi, Karnataka, India
6 Department of Oral and Maxillofacial Surgery, Maratha Mandal's N. G. Halgekar Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India

Correspondence Address:
Dr. Kishore G Bhat
Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Near KSRP Ground, Bauxite Road, Belagavi - 590 010, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ccd.ccd_252_20

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Objective: Cancer stem cells (CSCs) belong to a subpopulation of undifferentiated cells present within tumors that have the potential to regenerate, differentiate, maintenance of pluripotency, drug resistance, and tumorigenicity when transplanted into an innate host. These can influence the growth and behavior of these tumors and are used to investigate the initiation, progression, and treatment strategies of laryngeal cancer. Research on CSC science and targeted therapies were hinge on their isolation and/or enrichment procedures. The object of the study is to isolate cancer stem cells from primary laryngeal carcinoma (CSCPLC) by tumor spheres enrichment. We checked the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance. Materials and Methods: We performed tumor sphere formation assay (primary, secondary, and tertiary) chemotherapy resistance by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were performed to evaluate the CSC cells. Immunofluorescence for stem cell markers (CD133+, CD44+) and gene expression of stem cell markers for CD133+, CD44+, OCT4, SOX2, and NANOG was done using the real-time polymerase chain reaction technique. Results: We were able to isolated CSC subpopulations from PLC cell lines by the tumor sphere method. These cells exhibited good primary, secondary, and tertiary tumor sphere formation efficiency and also disclosed a resistant index of more than 2. Immunofluorescence for stem cell markers (CD133+ and CD44+) confirms the presence of CSC. There was significantly higher mRNA expression of stem cell markers in CSC enriched subpopulations compared to the parental cell lines. Conclusion: We conclude that tumor spheres enrichment is an efficient, economical, and reliable approach for the isolation and characterization of CSC from PLC cell lines. These cells demonstrated the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance.


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