小分子锌指蛋白1在4-硝基喹啉-N-氧化物诱导SD大鼠颊黏膜癌变中的表达研究

doi:10.7518/hxkq.2020.02.004

华西口腔医学杂志 ›› 2020, Vol. 38 ›› Issue (2): 133-138.doi: 10.7518/hxkq.2020.02.004

小分子锌指蛋白1在4-硝基喹啉-N-氧化物诱导SD大鼠颊黏膜癌变中的表达研究

余丽¹^,²^,³, 刘旭倩¹^,², 陈雨荷¹^,², 陈潇⁴, 聂敏海¹^,²()

^1. 西南医科大学口颌面修复重建与再生实验室,泸州 646000
^2. 西南医科大学附属口腔医院牙周黏膜科,泸州 646000
^3. 北海道大学齿学院口腔病态学分野口腔诊断内科学教室,日本北海道 060-0808
^4. 绵阳市口腔医院正畸科,绵阳 621000

收稿日期:2019-07-07 修回日期:2020-01-15 出版日期:2020-04-01 发布日期:2020-04-15
通讯作者: 聂敏海 E-mail:nieminhai@126.com
作者简介:余丽,住院医师,博士,E-mail： yulixt126@126.com
基金资助:
四川省医学会科研课题计划(S17071);四川省科技厅项目(2018JY040)

Study on expression of LIM domain only protein 1 in SD rat oral buccal mucosa carcinogenesis induced by 4-nitroquinoline N-oxide

Yu Li¹^,²^,³, Liu Xuqian¹^,², Chen Yuhe¹^,², Chen Xiao⁴, Nie Minhai¹^,²()

^1. Oral & Maxillofacial Reconstruction Laboratory, Southwest Medical University, Luzhou 646000, China
^2. Dept. of Periodontal Mucosal Disease, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China
^3. Dept. of Oral Diagnosis and Medicine, Division of Oral Pathobiological Science, Graduate School of Dental Medicine, Hokkaido University, Hokkaido 060-0808, Japan
^4. Dept. of Orthodontics, Mianyang Stomatological Hospital, Mianyang 621000, China

Received:2019-07-07 Revised:2020-01-15 Online:2020-04-01 Published:2020-04-15
Contact: Minhai Nie E-mail:nieminhai@126.com
Supported by:
Sichuan Medical Association Scientific Research Project(S17071);Sichuan Science and Technology Department Project(2018JY040)

摘要/Abstract

摘要：

目的研究口腔黏膜癌变中小分子锌指蛋白1（LMO1）在基因转录和蛋白水平的表达变化。方法取本课题组前期4-硝基喹啉-N-氧化物（4NQO）饮水法构建鳞癌动物模型的49例样本进行苏木精-伊红（HE）染色,依据上皮异常增生程度确定实验组病理分级并确定实验分组;免疫组织化学染色定性确定LMO1的表达部位;实时荧光定量聚合酶链式反应（RT-qPCR）和Western blot检测5组样本中LMO1 mRNA和蛋白的表达。结果 HE染色确定对照组7例,轻度组6例,中度组11例,重度组9例,OSCC组16例。免疫组织化学染色检测可见,LMO1主要表达于细胞质,对照组、轻度组、中度组、重度组和OSCC组的阳性表达率分别为14.3%、33.3%、81.8%、88.9%、93.8%。RT-qPCR检测可见,对照组LMO1 mRNA的表达量最低,OSCC组最高,对照组与轻度组间mRNA表达差异无统计学意义（P>0.05）,其余各组组间两两比较,mRNA的表达差异均有统计学意义（P<0.05）。Western blot检测可见,随着上皮异常增生程度的加重,LMO1的蛋白表达量逐渐上升,在OSCC组中表达最高。结论口腔黏膜癌变进程中,LMO1 mRNA和蛋白异常表达,且mRNA和蛋白表达量随上皮异常增生程度的加重而增加。

关键词: 小分子锌指蛋白1, 口腔鳞状细胞癌, 上皮异常增生

Abstract:

Objective This work aimed to determine the expression changes in LIM domain only protein 1 (LMO1) in gene transcription and protein levels during oral squamous cell carcinoma (OSCC) development. Methods The tissues in this study were taken from our team’s previous animal model building, and we performed hematoxylin-eosin (HE) staining on 49 cases. The pathological classification of the experiment group was determined on the basis of the abnormal epithelial hyperplasia degree. The expression part of LMO1 was determined by immunohistochemistry staining. The mRNA and protein LMO1 expression levels in five groups were detected by real-time fluorescent quantitative of nucleotide polymer chain reaction (RT-qPCR) and Western blot, respectively. Results HE staining determined 7 cases of the control group, 6 cases of mild epithelial dysplasia, 11 cases of moderate epithelial dysplasia, 9 cases of severe epithelial dysplasia, and 16 cases of OSCC. Immunohistochemistry results: LMO1 expression was localized in the cytoplasm, and the positive expression rates of the protein LMO1 in the control and experiment groups were 14.3% for normal buccal mucosal tissue, 33.3% for mild epithelial dysplasia, 81.8% for moderate epithelial dysplasia, 88.9% for severe epithelial dysplasia, and 93.8% for OSCC. RT-qPCR results: mRNA expression was lowest in the control group and highest in the OSCC group, the difference between the mild dysplasia and control groups was not significant (P>0.05). Pairwise comparison among other groups showed statistically significant differences (P<0.05). Western blot results: with the aggravation of the pathological degree, the protein LMO1 expression level increased gradually. The OSCC group expressed the highest LMO1 expression level. Conclusion The oral mucosa carcinogenesis models showed abnormal the mRNA and protein LMO1 expression levels, and the mRNA and protein expression levels were positively correlated with the degree of abnormal proliferation.

Key words: LIM domain only protein 1, oral squamous cell carcinoma, epithelial dysplasia

中图分类号:

R781.9

余丽, 刘旭倩, 陈雨荷, 陈潇, 聂敏海. 小分子锌指蛋白1在4-硝基喹啉-N-氧化物诱导SD大鼠颊黏膜癌变中的表达研究[J]. 华西口腔医学杂志, 2020, 38(2): 133-138.

Yu Li, Liu Xuqian, Chen Yuhe, Chen Xiao, Nie Minhai. Study on expression of LIM domain only protein 1 in SD rat oral buccal mucosa carcinogenesis induced by 4-nitroquinoline N-oxide[J]. West China Journal of Stomatology, 2020, 38(2): 133-138.

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组别	n	LMO1的表达				阳性率/%	Z值	P值
组别	n	-	+	++	+++	阳性率/%	Z值	P值
对照	7	6	1	0	0	14.3	-	-
轻度	6	4	1	1	0	33.3	-0.781	0.435
中度	11	2	2	5	2	81.8	-2.836	0.005
重度	9	1	1	3	4	88.9	-3.020	0.003
OSCC	16	1	3	3	9	93.8	-3.545	0.000

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小分子锌指蛋白1在4-硝基喹啉-N-氧化物诱导SD大鼠颊黏膜癌变中的表达研究

Study on expression of LIM domain only protein 1 in SD rat oral buccal mucosa carcinogenesis induced by 4-nitroquinoline N-oxide

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