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微核试验:一种评估烟草使用者、口腔白斑病患者和口腔黏膜下纤维化患者遗传毒性变化的早期诊断工具。

Micronucleus Assay: An Early Diagnostic Tool to Assess Genotoxic Changes in Patients with Tobacco Use, Oral Leukoplakia and Oral Submucous Fibrosis.

作者信息

Kohli Moulshree, Ahuja Puneet, Mehendiratta Monica, Sharma Mohit, Dutta Jahnobi

机构信息

Senior Lecturer, Department of Oral Pathology, I.T.S. Dental College, Hospital and Research Centre, Greater Noida, Uttar Pradesh, India.

Principal, Professor and Head, Department of Oral Pathology, I.T.S. Dental College, Hospital and Research Centre, Greater Noida, Uttar Pradesh, India.

出版信息

J Clin Diagn Res. 2017 Sep;11(9):ZC28-ZC32. doi: 10.7860/JCDR/2017/27711.10567. Epub 2017 Sep 1.

Abstract

INTRODUCTION

Micronuclei (MNi) are acentric chromatid or chromosome fragments produced via genetic damage through genotoxic agents contained in tobacco and betel nut. Evidently, the various Oral Potentially Malignant Disorders (OPMDs) like oral lichen Planus, oral leukoplakia and Oral Submucous Fibrosis (OSMF) demonstrate MNi, as a substantiation of genetic damage. As these changes can be easily appreciated in oral exfoliated cells, an exfoliated cell based MNi assay might be utilized as handy and non invasive biomonitoring tool for gauging the genetic damage and hence the propensity for malignant transformation in OPMDs. To this end, MNi are definitely easier to evaluate when compared to chromosome aberrations.

AIM

To compare the MNi frequency in normal mucosa, in individuals using various tobacco forms without oral leukoplakia, individuals using various tobacco forms with oral leukoplakia, and areca nut chewers with OSMF, using three different stains.

MATERIALS AND METHODS

Oral exfoliated cells from 50 cases of normal mucosa (Group I), 50 cases of tobacco chewing people without Oral Leukoplakia (Group II), 50 cases of people with Oral Leukoplakia (Group III) and 50 cases of areca nut chewers with OSMF (Group IV) were taken. MNi frequencies were compared in these groups using three different stains i.e., Papanicolaou (PAP) stain, May Grunwald Giemsa (MGG) stain and Feulgen stain. The data between cases (Group II, III and IV) and control groups (Group I) was analyzed by Kruskal-Wallis Test. The comparison between two independent groups was done by Mann-Whitney U test and interstain comparison between cases and control was done by Wilcoxon Signed Rank Test and the individual p-value was obtained.

RESULTS

A significant increase in the count was observed during transition of normal mucosa to OPMDs. The best stain for detecting MNi was PAP stain followed by MGG stain and Feulgen stain.

CONCLUSION

The higher mean MNi count for PAP stain and MGG stain could be attributed to nonspecific staining. Further study using a larger sample size on quantitative assessment of MNi count in various OPMDs is warranted.

摘要

引言

微核(MNi)是由烟草和槟榔中含有的基因毒性物质导致遗传损伤而产生的无着丝粒染色单体或染色体片段。显然,各种口腔潜在恶性疾病(OPMDs),如口腔扁平苔藓、口腔白斑和口腔黏膜下纤维化(OSMF),都表现出微核,这是遗传损伤的一种证明。由于这些变化在口腔脱落细胞中很容易观察到,基于脱落细胞的微核检测可能是一种方便且无创的生物监测工具,用于评估遗传损伤以及OPMDs中恶性转化的倾向。为此,与染色体畸变相比,微核肯定更容易评估。

目的

使用三种不同的染色剂,比较正常黏膜、使用各种烟草形式但无口腔白斑的个体、使用各种烟草形式且有口腔白斑的个体以及患有OSMF的槟榔咀嚼者的微核频率。

材料与方法

采集50例正常黏膜(I组)、50例咀嚼烟草但无口腔白斑的人(II组)、50例患有口腔白斑的人(III组)和50例患有OSMF的槟榔咀嚼者(IV组)的口腔脱落细胞。使用三种不同的染色剂,即巴氏(PAP)染色、美-格(MGG)染色和福尔根染色,比较这些组中的微核频率。病例组(II、III和IV组)与对照组(I组)之间的数据采用Kruskal-Wallis检验进行分析。两个独立组之间的比较采用Mann-Whitney U检验,病例组与对照组之间的染色剂间比较采用Wilcoxon符号秩检验,并获得个体p值。

结果

在从正常黏膜转变为OPMDs的过程中,观察到计数显著增加。检测微核的最佳染色剂是PAP染色,其次是MGG染色和福尔根染色。

结论

PAP染色和MGG染色的微核平均计数较高可能归因于非特异性染色。有必要使用更大样本量对各种OPMDs中的微核计数进行定量评估的进一步研究。

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