DNA甲基化转移酶1/3b与肝癌临床病理特征和预后相关性研究

doi:10.3877/cma.j.issn.1674-0793.2015.04.002

中华普通外科学文献(电子版) ›› 2015, Vol. 09 ›› Issue (04) : 261 -266. doi: 10.3877/cma.j.issn.1674-0793.2015.04.002

所属专题：文献；

论著

DNA甲基化转移酶1/3b与肝癌临床病理特征和预后相关性研究

方钦亮¹, 谢程融¹, 赵文秀¹, 孙红光¹, 尹毅锐¹, 张盛¹, 王付强¹, 彭友缘¹, 王效民¹, 尹震宇¹^,^†(

)

^1. 361004　厦门大学附属中山医院肝胆外科　福建省慢性肝病肝癌重点实验室

收稿日期:2015-02-05 出版日期:2015-08-01
通信作者: 尹震宇
基金资助:
国家自然科学基金资助项目(81172286)

Correlation between DNMT1/3b and the clinicopathology and prognosis of hepatocellular carcinoma

Qinliang Fang¹, Chengrong Xie¹, Wenxiu Zhao¹, Hongguang Sun¹, Yirui Yin¹, Sheng Zhang¹, Fuqiang Wang¹, Youyuan Peng¹, Xiaomin Wang¹, Zhenyu Yin¹^,^†()

^1. Department of Hepatobiliary Surgery, Zhongshan Hospital of Xiamen University, Fujian Provincial Key Laboratory of Chronic Liver Disease and Hepatoce- llular Carcinoma, 361004 Xiamen, China

Received:2015-02-05 Published:2015-08-01
Corresponding author: Zhenyu Yin
About author:
Corresponding author: Yin Zhenyu, Email: yinzy@xmu.edu.cn

全文 ( ) 下载PDF ( ) 导出引用

引用本文：

方钦亮, 谢程融, 赵文秀, 孙红光, 尹毅锐, 张盛, 王付强, 彭友缘, 王效民, 尹震宇. DNA甲基化转移酶1/3b与肝癌临床病理特征和预后相关性研究[J/OL]. 中华普通外科学文献(电子版), 2015, 09(04): 261-266.

Qinliang Fang, Chengrong Xie, Wenxiu Zhao, Hongguang Sun, Yirui Yin, Sheng Zhang, Fuqiang Wang, Youyuan Peng, Xiaomin Wang, Zhenyu Yin. Correlation between DNMT1/3b and the clinicopathology and prognosis of hepatocellular carcinoma[J/OL]. Chinese Archives of General Surgery(Electronic Edition), 2015, 09(04): 261-266.

目的

探讨DNA甲基化转移酶1/3b(DNMT1/3b)表达异常与肝癌临床病理特征和患者术后生存之间的相关性。

方法

采用免疫组织化学技术检测89例手术切除肝癌组织中DNMT1和DNMT3b的蛋白表达情况,结合临床病理及无瘤生存期和总生存期,分析DNMT1/3b与临床病理参数、预后的相关性,并通过高通量基因芯片技术筛选肝癌相关DNMT1和DNMT3b的靶基因。

结果

89例肝癌组织中DNMT1和DNMT3b皆为阳性63例,皆为阴性5例,单一阳性的分别为9例和12例。DNMT1/3b在肝癌组织中高表达与患者血清AFP(χ²=12.903,P=0.005)、乙型肝炎(χ²=9.535,P=0.023)、卫星灶(χ²=9.574,P=0.023)和肿瘤复发、转移及生存期有关,而与性别、肿瘤大小、肝硬化和肿瘤分化无关;Kaplan-Meier分析显示DNMT1/3b阳性组患者生存时间较阴性组短,差异有统计学意义(P<0.05)。高通量基因组甲基化芯片技术筛选出肝癌细胞中受DNMT1和DNMT3b调控的基因2 000多个,其中参与肿瘤侵袭转移的靶基因有112个。

结论

DNMT1/3b在肝癌组织中高表达与患者血清AFP、乙型肝炎、卫星灶和肿瘤复发、转移等临床病理特征和生存期密切相关,并可能通过调控多方面肿瘤相关基因促进肝癌复发、转移等恶性表型。

关键词: 肝癌, DNA甲基化转移酶, 生存期, 甲基化芯片

Objective

To detect the protein expression of DNMT1 and DNMT3b, and to explore the association of DNMT1/3b with the clinicopathology and prognosis of hepatocellular carcinoma.

Methods

Eighty-nine cases of hepatocellular carcinoma were detected by immunohistochemistry (IHC) for analysis of the correlation between DNMT1/3b expression and clinicopathologic parameters and the tumor-free survival and overall survival. And high-throughput gene chip technology was used to screen out DNMT1 and DNMT3b target genes.

Results

Both DNMT1 and DNMT3b expressions existed in 63 cases while 5 cases had neither DNMT1 nor DNMT3b. Nine cases had positive DNMT1 and 12 had positive DNMT3b. High level expression of DNMT1/3b was associated with serum AFP (χ²=12.903, P=0.005), HBV (χ²=9.535, P=0.023), satellite opacities (χ²=9.574, P=0.023), tumor recurrence and metastasis, and overall survival. No association was found with sex, size of tumor, liver cirrhosis and tumor differentiation. By Kaplan-Meier analysis, survival time was shorter in the patients with DNMT1 and/or 3b overexpression than in those with negative DNMT1 and 3b. More than 2 000 DNMT1 and DNMT3b target genes were screened out through high-throughput gene chip technology, and 112 of which were involved in tumor invasion and metastasis.

Conclusions

The overexpression of DNMT1 and DNMT3b are significantly related to the clinical and pathological characteristics such as serum AFP, HBV, satellite opacities, tumor recurrence and metastasis, and survival rate in hepatocellular carcinoma, and they can regulate a various tumor related genes in promoting the malignant phenotype including recurrence and metastasis of hepatocellular carcinoma.

Key words: Hepatocellular carcinoma, DNA methyltransferases, Survival rate, Methylation microarrays

表1 DNNT1和DNMT3b蛋白表达与肝癌患者临床病理特征的关系(例)

临床病理特征		DNMT1/3b蛋白表达(例数)				χ²值	P值
临床病理特征		(-/-)	(+/-)	(-/+)	(+/+)	χ²值	P值
性别		?	?	?	?	0.469	0.926
?	男	4	8	11	55	?	?
?	女	1	1	1	8	?	?
HBsAg		?	?	?	?	9.535	0.023
?	阳性	2	9	8	53	?	?
?	阴性	3	0	4	10	?	?
血清AFP(μg/L)		?	?	?	?	12.903	0.005
?	<25	5	2	5	16	?	?
?	≥25	0	7	7	47	?	?
肿瘤大小(cm)		?	?	?	?	0.167	0.983
?	<5	2	4	5	24	?	?
?	≥5	3	5	7	39	?	?
肝硬化		?	?	?	?	1.053	0.788
?	阳性	2	4	6	36	?	?
?	阴性	3	5	6	27	?	?
肿瘤分化		?	?	?	?	2.491	0.870
?	低	1	2	2	8	?	?
?	中	3	5	9	50	?	?
?	高	1	2	1	5	?	?
卫星灶		?	?	?	?	9.574	0.023
?	阳性	0	1	1	7	?	?
?	阴性	5	8	11	57	?	?

表1 DNNT1和DNMT3b蛋白表达与肝癌患者临床病理特征的关系(例)

临床病理特征		DNMT1/3b蛋白表达(例数)				χ²值	P值
临床病理特征		(-/-)	(+/-)	(-/+)	(+/+)	χ²值	P值
性别		?	?	?	?	0.469	0.926
?	男	4	8	11	55	?	?
?	女	1	1	1	8	?	?
HBsAg		?	?	?	?	9.535	0.023
?	阳性	2	9	8	53	?	?
?	阴性	3	0	4	10	?	?
血清AFP(μg/L)		?	?	?	?	12.903	0.005
?	<25	5	2	5	16	?	?
?	≥25	0	7	7	47	?	?
肿瘤大小(cm)		?	?	?	?	0.167	0.983
?	<5	2	4	5	24	?	?
?	≥5	3	5	7	39	?	?
肝硬化		?	?	?	?	1.053	0.788
?	阳性	2	4	6	36	?	?
?	阴性	3	5	6	27	?	?
肿瘤分化		?	?	?	?	2.491	0.870
?	低	1	2	2	8	?	?
?	中	3	5	9	50	?	?
?	高	1	2	1	5	?	?
卫星灶		?	?	?	?	9.574	0.023
?	阳性	0	1	1	7	?	?
?	阴性	5	8	11	57	?	?

图1 免疫组织化学法检测89例HCC患者手术切除组织(包括癌组织T及癌旁组织NT)中DNMT1/3b的表达（×200）

图2 HCC患者DNMT1/DNMT3b的表达与预后的相关性分析

表2 112个肿瘤侵袭、转移相关基因中甲基化程度最高的前20个基因

染色体	甲基化得分	基因名称	基因库编码	编码蛋白名称
chr15	4.59	CTSH	NM_004390	cathepsin H isoform a preproprotein
chr1	3.74	FOXE3	NM_012186	forkhead box E3
chr15	3.71	TYRO3	NM_006293	TYRO3 protein tyrosine kinase precursor
chr20	3.70	PARD6B	NM_032521	PAR-6 beta
chr3	3.63	PODXL2	NM_015720	podocalyxin-like 2 precursor
chr16	3.63	SOX8	NM_014587	SRY (sex determining region Y)-box 8
chr17	3.61	ABR	NM_021962	active breakpoint cluster region-related
chrX	3.60	SHROOM2	NM_001649	apical protein of Xenopus-like
chr8	3.52	DLC1	NM_006094	deleted in liver cancer 1 isoform 2
chr8	3.52	PTK2B	NM_004103	PTK2B protein tyrosine kinase 2 beta isoform a
chr4	3.49	PITX2	NM_153427	paired-like homeodomain transcription factor 2
chr6	3.44	SRF	NM_003131	serum response factor (c-fos serum response
chr11	3.39	P2RY2	NM_176071	purinergic receptor P2Y2
chr2	3.35	NCK2	NM_003581	NCK adaptor protein 2 isoform A
chr17	3.28	ACE	NM_000789	angiotensin I converting enzyme 1 isoform 1
chr7	3.23	FSCN1	NM_003088	fascin 1
chr1	3.22	ARHGEF16	NM_014448	Rho guanine exchange factor 16
chr17	3.20	CDK5R1	NM_003885	cyclin-dependent kinase 5, regulatory subunit 1
chr9	3.19	NTRK2	NM_001007097	neurotrophic tyrosine kinase, receptor, type 2
chr20	3.14	SOX18	NM_018419	SRY-box 18

表2 112个肿瘤侵袭、转移相关基因中甲基化程度最高的前20个基因

染色体	甲基化得分	基因名称	基因库编码	编码蛋白名称
chr15	4.59	CTSH	NM_004390	cathepsin H isoform a preproprotein
chr1	3.74	FOXE3	NM_012186	forkhead box E3
chr15	3.71	TYRO3	NM_006293	TYRO3 protein tyrosine kinase precursor
chr20	3.70	PARD6B	NM_032521	PAR-6 beta
chr3	3.63	PODXL2	NM_015720	podocalyxin-like 2 precursor
chr16	3.63	SOX8	NM_014587	SRY (sex determining region Y)-box 8
chr17	3.61	ABR	NM_021962	active breakpoint cluster region-related
chrX	3.60	SHROOM2	NM_001649	apical protein of Xenopus-like
chr8	3.52	DLC1	NM_006094	deleted in liver cancer 1 isoform 2
chr8	3.52	PTK2B	NM_004103	PTK2B protein tyrosine kinase 2 beta isoform a
chr4	3.49	PITX2	NM_153427	paired-like homeodomain transcription factor 2
chr6	3.44	SRF	NM_003131	serum response factor (c-fos serum response
chr11	3.39	P2RY2	NM_176071	purinergic receptor P2Y2
chr2	3.35	NCK2	NM_003581	NCK adaptor protein 2 isoform A
chr17	3.28	ACE	NM_000789	angiotensin I converting enzyme 1 isoform 1
chr7	3.23	FSCN1	NM_003088	fascin 1
chr1	3.22	ARHGEF16	NM_014448	Rho guanine exchange factor 16
chr17	3.20	CDK5R1	NM_003885	cyclin-dependent kinase 5, regulatory subunit 1
chr9	3.19	NTRK2	NM_001007097	neurotrophic tyrosine kinase, receptor, type 2
chr20	3.14	SOX18	NM_018419	SRY-box 18

图3 肝癌细胞表型相关特点中受DNMT1/3b调控的基因分类数量

[1]	Jemal A, Bray F, Center MM, et al. Global cancer statistics[J]. CA Cancer J Clin, 2011, 61(2): 69-90.
[2]	Park JY. Promoter hypermethylation as a biomarker in prostate adenocarcinoma[J]. Methods Mol Biol, 2015, 1238: 607-625.
[3]	Ng JM, Yu J. Promoter hypermethylation of tumour suppressor genes as potential biomarkers in colorectal cancer[J]. Int J Mol Sci, 2015, 16(2): 2472-2496.
[4]	Wang D, Yang PN, Chen J, et al. Promoter hypermethylation may be an important mechanism of the transcriptional inactivation of ARRDC3, GATA5, and ELP3 in invasive ductal breast carcinoma[J]. Mol Cell Biochem, 2014, 396(1-2): 67-77.
[5]	Kang GH. CpG island hypermethylation in gastric carcinoma and its premalignant lesions[J]. Korean J Pathol, 2012, 46(1): 1-9.
[6]	Lin RK, Hsu HS, Chang JW, et al. Alteration of DNA methyltransferases contributes to 5′CpG methylation and poor prognosis in lung cancer[J]. Lung Cancer, 2007, 55(2): 205-213.
[7]	Oh BK, Kim H, Park HJ, et al. DNA methyltransferase expression and DNA methylation in human hepatocellular carcinoma and their clinicopathological correlation[J]. Int J Mol Med, 2007, 20(1): 65-73.
[8]	Fang QL, Yin YR, Xie CR, et al. Mechanistic and biological significance of DNA methyltransferase 1 upregulated by growth factors in human hepatocellular carcinoma[J]. Int J Oncol, 2015, 46(2): 782-790.
[9]	Fan H, Zhao ZJ, Cheng J, et al. Overexpression of DNA methyltransferase 1 and its biological significance in primary hepatocellular carcinoma[J]. World J Gastroenterol, 2009, 15(16): 2020-2026.
[10]	Saito Y, Kanai Y, Nakagawa T, et al. Increased protein expression of DNA methyltransferase (DNMT) 1 is significantly correlated with the malignant potential and poor prognosis of human hepatocellular carcinomas[J]. Int J Cancer, 2003, 105(4): 527-532.
[11]	Shitani M, Sasaki S, Akutsu N, et al. Genome-wide analysis of DNA methylation identifies novel cancer-related genes in hepatocellular carcinoma[J]. Tumour Biol, 2012, 33(5): 1307-1317.
[12]	Yu J, Ni M, Xu J, et al. Methylation profiling of twenty promoter-CpG islands of genes which may contribute to hepatocellular carcinogenesis[J]. BMC Cancer, 2002, 2: 29.

[1]	丁科, 张亚琼, 刘杰, 邓莉平, 张永喜, 熊勇. 获得性免疫缺陷综合征相关淋巴瘤患者的临床特征及生存状况的变化趋势[J/OL]. 中华实验和临床感染病杂志(电子版), 2024, 18(05): 278-284.
[2]	唐梅, 周丽, 牛岑月, 周小童, 王倩. ICG荧光导航的腹腔镜肝切除术临床意义[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(06): 655-658.
[3]	高金红, 陈玉梅, 郭韵. 基于King互动达标理论的心理疏导在腹腔镜肝癌切除术患者的应用效果分析[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(05): 517-520.
[4]	林巧, 周丽. RFA联合LAH术治疗原发性肝癌并门静脉癌栓的临床效果分析[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(05): 521-524.
[5]	曾繁利, 齐秩凯, 杨贺庆. 两种经Glisson蒂鞘解剖路径肝切除术治疗原发性肝癌的肿瘤学疗效及风险比对[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(05): 525-527.
[6]	胡宁宁, 赵延荣, 王栋, 王胜亮, 郭源. FMNL3与肝细胞癌肝移植受者预后的相关性研究[J/OL]. 中华移植杂志(电子版), 2024, 18(05): 283-288.
[7]	张礼江, 沈玲佳, 施我大. 倾向性评分匹配分析奥希替尼对晚期NSCLC 预后的影响[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(05): 820-822.
[8]	王飞, 陈政, 余德才, 曹亚娟. 原发性肝癌合并门静脉高压症的微创手术治疗[J/OL]. 中华腔镜外科杂志(电子版), 2024, 17(05): 306-310.
[9]	张敏, 朱建华, 缪雅芳, 郭锦荣. 菝葜皂苷元对肝癌HepG2细胞抑制作用的机制研究[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(06): 328-335.
[10]	曾聿理, 雷发容, 肖慧, 邱德亮, 谢静, 吴寻. 氯普鲁卡因通过调控circRNA-ZKSCAN1表达抑制肝癌Huh-7细胞体外生长和转移的研究[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 220-228.
[11]	袁雨涵, 杨盛力. 体液和组织蛋白质组学分析在肝癌早期分子诊断中的研究进展[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 883-888.
[12]	吴警, 吐尔洪江·吐逊, 温浩. 肝切除术前肝功能评估新进展[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 889-893.
[13]	广东省护士协会介入护士分会, 广东省医师协会介入医师分会. 原发性肝癌低血糖患者护理规范管理专家共识[J/OL]. 中华临床医师杂志(电子版), 2024, 18(08): 709-714.
[14]	韦巧玲, 黄妍, 赵昌, 宋庆峰, 陈祖毅, 黄莹, 蒙嫦, 黄靖. 肝癌微波消融术后中重度疼痛风险预测列线图模型构建及验证[J/OL]. 中华临床医师杂志(电子版), 2024, 18(08): 715-721.
[15]	蔡晓雯, 李慧景, 丘婕, 杨翼帆, 吴素贤, 林玉彤, 何秋娜. 肝癌患者肝动脉化疗栓塞术后疼痛风险预测模型的构建及验证[J/OL]. 中华临床医师杂志(电子版), 2024, 18(08): 722-728.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Correlation between DNMT1/3b and the clinicopathology and prognosis of hepatocellular carcinoma

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Correlation between DNMT1/3b and the clinicopathology and prognosis of hepatocellular carcinoma