光动力治疗对头颈部鳞状细胞癌细胞周期蛋白D1和P53 mRNA表达的影响

M. Koopaie, Mahsan Mosaieby, Zahra Jabbarpour, A. Shamshiri
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引用次数: 0

摘要

背景:光动力疗法(PDT)被认为是头颈部鳞状细胞癌(HNSCC)的替代治疗方法之一。P53 mRNA是一种肿瘤抑制基因,其突变增加了不受控制的癌样分裂的可能性。致癌基因CCND1的突变增加了作为致瘤蛋白的细胞周期蛋白D1的产生。目的:本研究旨在确定甲苯胺蓝光敏剂PDT对HNSCC细胞系CCND1和P53 mRNA水平的影响。方法:从NCBI提取人HNSCC细胞系。采用C196编号HN5。将细胞分为四组:I组(甲苯胺蓝和激光照射作用下的HNSCC细胞系)、II组(甲苯胺蓝作用下的HNSCC细胞系)、III组(激光照射下的HNSCC细胞系)、IV组(对照组,HNSCC细胞系)。以甲苯胺蓝为光敏剂的660nm THOR激光器。RNA提取分为三个步骤,包括细胞降解、纯化和酒精沉淀。cDNA的制备采用Takara Kit。采用扩增试剂盒和Real-Time PCR检测mRNA相对水平。结果:ⅰ组P53 mRNA相对水平为1.50±0.33 (P = 0.65),ⅱ组为1.49±0.23 (P =0.5),ⅲ组为1.40±1.05 (P = 0.63)。与对照组比较,ⅰ组CCND1 mRNA平均升高18.01±3.37 (p=0.04),ⅱ组17.69±3.3 (p= 0.03),ⅲ组9.01±6.17 (p= 0.20)。结论:通过单因素方差分析比较P53、CCND1 mRNA的折线变化指数,结果显示,治疗组与对照组相比,P53、CCND1 mRNA表达增加,但两组间P53 mRNA (P = 0.99)和CCND1 mRNA (P = 0.22)指数升高无统计学意义。本研究结果可以为更好地理解HNSCC细胞系PDT的基因机制提供一个起点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Effect of Photodynamic Therapy on Cyclin D1 and P53 mRNA Levels in Head and Neck Squamous Cell Carcinoma Cell Line
Background: Photodynamic Therapy (PDT) is considered as one of the alternative treatments for Head and Neck Squamous Cell Carcinoma (HNSCC). P53 mRNA is a tumor suppressor gene whose mutation increases the likelihood of uncontrolled cancer-like divisions. The mutation of CCND1 oncogenic increases the production of cyclin D1 as a tumorigenic protein. Objectives: This study aimed to determine the effect of PDT using toluidine blue as a photosensitizer on the CCND1 and P53 mRNA levels in the HNSCC cell line. Methods: A human HNSCC cell line from NCBI.C196 designation HN5 was used. Cells were divided into four groups: Group I (HNSCC cell line under the influence of toluidine blue and laser irradiation), group II (HNSCC cell line under the influence of toluidine blue), group III (HNSCC cell line under laser irradiation), group IV (control group, HNSCC cell line). A 660 nm THOR laser using toluidine blue as a photosensitizer was applied. The RNA extraction was performed in three steps, including cell degradation, purification, and precipitation by alcohol. The cDNA was prepared using Takara Kit. The Amplicon kit and Real-Time PCR analysis were used for the assessment of mRNA relative levels. Results: The P53 mRNA relative levels were 1.50 ± 0.33 in group I (P = 0.65), 1.49 ± 0.23 (P =0.5) in group II, and 1.40 ± 1.05 (P = 0.63) in group III. Compared to the control group, the mean increases in CCND1 mRNA were 18.01 ± 3.37 (p=0.04) in group I, 17.69 ± 3.3 (P = 0.03) in group II, and 9.01 ± 6.17 (P = 0.20) in group III. Conclusions: The comparison of the fold change index for P53 and CCND1 mRNA by the one-way ANOVA test showed that despite the increased expression of P53 and CCND1 mRNA in treatment groups compared to the control group, there was no statistically significant difference between the increases in P53 mRNA (P = 0.99) and CCND1 mRNA (P = 0.22) index between the groups. The results of this study could be a starting point for a better understanding of the mechanism of genes in PDT of the HNSCC cell line.
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