突变分析揭示宫颈癌P53基因外显子3和外显子4的多态性变异

Olabode E. Omotoso, A. Gbadegesin Michael, A. O. Oluwasola Timothy, A. Okolo Clement, O. Ogun Gabriel, O. Oluwasola Abideen
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引用次数: 0

摘要

本研究探讨了宫颈癌中P53基因外显子3和外显子4的突变。从伊巴丹大学学院医院报告的患者的宫颈组织活检中分离和纯化了基因组DNA。用NanoDropTM 2000分光光度计对纯化的基因组DNA进行定量。用聚合酶链式反应(PCR)扩增感兴趣的区域,在1.5%琼脂糖凝胶上定量,并在透照器上观察溴化乙啶染色凝胶。PCR产物在Inqaba Biotec测序,使用FinchTv和SnapGene Viewer进行色谱分析。对DNA序列进行核苷酸BLAST鉴定,检索参考序列。使用T-COFFEE进行多序列比对以揭示样品中的多态性变异。本研究结果显示19个已确定的多态性变异;47%的样本发生错义突变,32%为沉默突变,16%为移码突变,5%为无义突变。社会人口学特征显示,60%的研究参与者的丈夫有多个性伴侣,只有23.3%的参与者在诊断前做过巴氏涂片检查,而其中20%的人不知道筛查测试。组织病理学结果和多重序列比对显示,随着宫颈癌严重程度的增加,p53基因倾向于积累更多的突变。我们的研究还揭示了尼日利亚妇女对癌症筛查知识的贫乏,这可能是非洲人口中宫颈癌诊断晚的因素之一。因此,鼓励对癌症筛查和早期发现的认识。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Mutational Analysis Revealed Polymorphic Variants in Exon 3 and Exon 4 Of P53 Gene in Cervical Cancer
This research study explored the mutations in the exon 3 and exon 4 of the P53 gene in cervical cancer. Genomic DNA was isolated and purified from cervical tissue biopsies obtained from patients reporting at the University College Hospital, Ibadan. The purified genomic DNA was quantified using NanoDropTM 2000 spectrophotometer. The region of interest was amplified using Polymerase Chain Reaction (PCR), this was quantified on a 1.5% agarose gel, and the ethidium bromide stained gel was viewed on a transilluminator. The PCR products were sequenced at Inqaba Biotec, and chromatogram was analyzed using FinchTv and SnapGene Viewer. Nucleotide BLAST was performed on the DNA sequence for sequence identity and retrieval of reference sequence. Multiple Sequence Alignment using T-COFFEE was performed to reveal the polymorphic variations in the samples. The outcome of this study showed nineteen identified polymorphic variants; Missense mutations occurred in 47% of the samples, 32% were silent mutations, 16% were frameshift mutation and 5% nonsense mutation. Sociodemographic characteristics revealed that 60% of the study participants has husbands with multiple sexual partners and that only 23.3% of the participants have ever done pap smear test prior to diagnosis, while 20% of them are unaware of the screening test. The histopathological result, alongside the Multiple Sequence Alignment revealed that with higher severity of cervical carcinoma, the p53 gene tends to accumulate more mutations. Our study also revealed the poor knowledge of Nigerian women to cancer screening, this may be one of the contributory factors to late diagnosis of cervical cancer among the African population. Hence, awareness for uptake of cancer screening and early detection are encouraged.
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