Effect of photodynamic therapy on expression of HRAS,NRAS and caspase 3 genes at mRNA levels,apoptosis of head and neck squamous cell carcinoma cell line |
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| Affiliation: | 1. Department of Oral Medicine, School of Dentistry, Tehran University of Medical Sciences, Tehran, Iran;2. Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran;3. Universal Scientific Education and Research Network (USERN), Tehran, Iran;4. Community Oral Health Department, School of Dentistry, Tehran University of Medical Sciences, Tehran, Iran;1. Department of Life Sciences, School of Pharmacy, International Medical University (IMU), Bukit Jalil, 57000, Kuala Lumpur, Malaysia;2. School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, 144411, Punjab, India;3. Faculty of Health, Australian Research Centre in Complementary and Integrative Medicine, University of Technology Sydney, Ultimo, 2007, Australia;4. Department of Chemistry, University of Petroleum & Energy Studies, Dehradun, 248007, India;5. Department of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Shoolini University, Solan, Himachal Pradesh, 173229, India;6. Department of Applied Chemistry, School of Applied Natural Science, Adama Science and Technology University, Po Box 1888, Adama, Ethiopia;7. School of Pharmacy, Suresh Gyan Vihar University, Mahal Road, Jagatpura, Jaipur, India;8. Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, India;9. Uttaranchal Institute of Pharmaceutical Sciences, Uttaranchal University, Dehradun, India;10. Department of Pharmaceutics, JSS College of Pharmacy, JSS Academy of Higher Education & Research, Ooty, Nilgiris, Tamil Nadu, India;11. Discipline of Pharmacy, Graduate School of Health, University of Technology Sydney, NSW, 2007, Australia;12. School of Life Sciences, University of Technology Sydney, Ultimo, NSW, 2007, Australia;13. Woolcock Institute of Medical Research, University of Sydney, Sydney, New South Wales, Australia;1. Centre for Healthcare Science and Technology, Indian Institute of Engineering, Science and Technology, Shibpur, Howrah, 711103, West-Bengal, India;2. Department of Oral and Maxillofacial Pathology, Buddha Institute of Dental Sciences, West TV Tower, Gandhinagar, Kankarabagh, Patna, 800020, Bihar, India;3. Department of Applied Mechanics, Indian Institute of Engineering, Science and Technology, Shibpur, Howrah, 711103, West-Bengal, India;4. Department of Computer Science and Technology, Indian Institute of Engineering, Science and Technology, Shibpur, Howrah, 711103, West-Bengal, India;2. Agricultural and Biological Engineering, Purdue University, West Lafayette, IN |
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| Abstract: | ObjectivesThis study aimed to assess the effect of photodynamic therapy (PDT) on expression of CASP3, NRAS and HRAS genes at mRNA levels, and apoptosis of head and neck squamous cell carcinoma (HNSCC) cell line.Materials and methodsIn order to complete the present in vitro study, HNSCC cell line (NCBI C196 HN5) purchased from Pasteur Institute. Cells were divided into four groups; Group 1: photodynamic treatment (laser + methylene blue (MB) as photosensitizer), group 2: MB, group 3: laser (with 660 nm wavelength), and group 4: control (without any treatment). To determine the optimal concentration of MB, in a pilot study, toxicity of MB in different concentration was assessed using MTT assay. Cells in group 1, 2 and 3 was treated at optimal concentration of MB (1.6 μg/mL). Gene expression at mRNA levels was assessed after 24 h incubation, using real-time (qRT)-PCR. The expression of BAX and BCL2 genes at the mRNA levels was analyzed to evaluate apoptosis. 2−ΔΔCt values of BCL2, BAX, CASP3, NRAS, and HRAS in groups was analyzed using ANOVA. Tukey’s HSD and Games Howell test was used to compare between two groups.ResultsOver-expression of BAX (p < 0.001), CASP3 (p < 0.001) and down-regulation of BCL2 (p = 0.004), HRAS (p = 0.023) and NRAS (p = 0.045) were noted in group 1 (PDT), compared with the control group. Treatment by laser alone induce down-regulation of CASP3 (p < 0.05), BAX (p < 0.05), BCL2 (p > 0.05), HRAS (p > 0.05) and NRAS (p > 0.05).ConclusionPDT caused down-regulation of NRAS, HRAS and BCL2 and over-expression of CASP3 and BAX genes at mRNA levels in HNSCC cell line. The present study raises the possibility that the role of MB on BCL2 down-regulation and BAX and CASP3 over-expression was higher than laser alone while it seems that laser alone was more effective than MB in HRAS and NRAS down-regulation. |
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| Keywords: | Head and neck squamous cell carcinoma Proto-oncogene Tumor suppressor genes Photodynamic therapy Apoptosis Methylene blue |
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