乙型肝炎病毒x蛋白介导微RNA-143在乙型肝炎相关肝癌患者中的表达及临床作用

doi:10.3877/cma.j.issn.2095-5820.2022.04.006

目的

探讨乙型肝炎（乙肝）相关肝病患者血清中乙型肝炎病毒x（HBx）蛋白与微RNA-143（miR-143）的相关性及在乙肝相关肝癌中的临床诊断价值。

方法

收集乙肝相关肝病患者血清169例，按乙肝病程进展分为慢性乙肝组78例、乙肝相关肝硬化组53例及乙肝相关肝癌组38例，同时收集20例健康人群血清设为正常对照组。采用酶联免疫吸附法（ELISA）测定血清HBx蛋白及乙肝两对半指标；实时荧光定量PCR（qPCR）检测血清miR-143及HBV DNA含量；全自动化学发光免疫分析仪（CLIA）检测血清甲胎蛋白（AFP）含量。

结果

miR-143与AFP表达量在HBx蛋白阳性组的表达水平均高于HBx蛋白阴性组，差异有统计学意义（P＜0.05）；HBx蛋白阳性检出率及miR-143表达量在HBV DNA高水平组较HBV DNA正常组、低水平组和中水平组的高，差异具有统计学意义（P＜0.05）；HBx蛋白阳性检出率在肝硬化组和肝癌组明显高于正常对照组和慢性乙肝组，血清miR-143及AFP表达量在肝癌患者中显著高于正常对照组、慢性乙肝组和肝硬化组，差异具有统计学意义（P＜0.05）；miR-143相对表达量在HBx蛋白阳性肝癌患者组中明显高于HBx蛋白阴性肝癌患者组，差异具有统计学意义（P＜0.05）；血清miR-143与HBx蛋白水平呈正相关（P＜0.05）。

结论

HBx蛋白与miR-143表达密切相关，呈一种剂量依赖关系，miR-143可能参与了HBx蛋白介导的乙肝肝癌进展的过程，两者在慢性乙肝发展成肝癌的进程中起着重要的作用，为乙肝肝癌诊疗提出一种新的思考方向。

关键词: 乙型肝炎病毒x蛋白, 微RNA-143, 乙肝相关肝癌

Objective

To study the expression of HBx protein (Hepatitis B virus X antigen,HBx)and miR-143 in the serum of patients with hepatitis B related liver disease, and to explore the relationship between HBx protein and miR-143, and their roles in hepatitis B related liver cancer.

Methods

169 serum samples were collected from patients with HBV-related liver disease in the Laboratory Department of the Second Affiliated Hospital of Guangzhou Medical University. According to the course of hepatitis B disease, they were divided into chronic hepatitis B group (78 cases), HBV-related liver cirrhosis group (53 cases) and HBV-related liver cancer group (38 cases). Serum samples from 20 healthy people were selected as the control group. Serum HBx protein and hepatitis B were detected by ELISA; Serum miR-143 and HBV DNA were detected by Quantitative Real-time PCR(qPCR); AFP was detected by automatic chemiluminescence immunoassay (CLIA).

Results

The expression levels of miR-143 and AFP in HBx positive group were significantly higher than those in HBx negative group (P＜0.05); The positive rate of HBx and the expression of miR-143 in the high level group of hepatitis B virus replication were significantly higher than those in the other three groups (P＜0.05); The positive rate of HBx in liver cirrhosis group and liver cancer group was significantly higher than that in chronic hepatitis B group. The expression of serum miR-143 and AFP in liver cancer patients was significantly higher than that in the other three groups (P＜0.05); The relative expression of miR-143 in HBx positive group was significantly higher than that in HBx negative group (P＜0.05).

Conclusions

serum HBx is closely related to the expression of miR-143 in a dose-dependent manner. miR-143 may be involved in the progression of hepatitis B virus X protein mediated hepatitis B liver cancer. Both of them play an important role in the development of chronic hepatitis B into liver cancer, which provides a new direction for the diagnosis and treatment of hepatitis B liver cancer.

Key words: HBx protein, miR-143, Hepatitis B related liver cancer

表1 HBx蛋白血清HBx蛋白、miR-143表达量与HBV DNA复制水平之间的分析

HBV DNA拷贝数（copies/ml）	例数	HBxAg		miR-143（2^–ΔΔCt值）
HBV DNA拷贝数（copies/ml）	例数	阳性［例（%）］	A_{450 mm}值中位数（min，max）	miR-143（2^–ΔΔCt值）
正常对照组	7	1（14.3）	0.34（0.017，0.80）	1.34±0.52
低水平组	66	14（21.2）	0.56（0.016，1.49）	2.23±0.78
中水平组	54	18（33.3）	0.96（0.031，2.02）	3.82±0.94
高水平组	42	22（52.4）^a	1.48（0.023，2.96）^a	4.64±0.87^a

表1 HBx蛋白血清HBx蛋白、miR-143表达量与HBV DNA复制水平之间的分析

HBV DNA拷贝数（copies/ml）	例数	HBxAg		miR-143（2^–ΔΔCt值）
HBV DNA拷贝数（copies/ml）	例数	阳性［例（%）］	A_{450 mm}值中位数（min，max）	miR-143（2^–ΔΔCt值）
正常对照组	7	1（14.3）	0.34（0.017，0.80）	1.34±0.52
低水平组	66	14（21.2）	0.56（0.016，1.49）	2.23±0.78
中水平组	54	18（33.3）	0.96（0.031，2.02）	3.82±0.94
高水平组	42	22（52.4）^a	1.48（0.023，2.96）^a	4.64±0.87^a

图1 HBx蛋白不同模式中miR-143与AFP表达量的分析

表2 HBx蛋白血清HBx蛋白、miR-143及AFP与乙肝病程关系的分析

组别	例数	HBx蛋白［例（%）］	miR-143（2^–ΔΔCt值）	HBV DNA（lg）	AFP（lg ug/L）
慢性乙肝组	78	10（12.8）	1.21±0.58	4.78±1.25	1.09±0.25
肝硬化组	53	27（50.8）^a	2.37±0.97	4.55±0.77	2.44±0.38
肝癌组	38	18（47.4）^a	4.23±1.08^b	5.53±1.18^b	4.74±0.57^b
正常对照组	20	0（0.0）	1.53±0.36	0	1.03±0.33

表2 HBx蛋白血清HBx蛋白、miR-143及AFP与乙肝病程关系的分析

组别	例数	HBx蛋白［例（%）］	miR-143（2^–ΔΔCt值）	HBV DNA（lg）	AFP（lg ug/L）
慢性乙肝组	78	10（12.8）	1.21±0.58	4.78±1.25	1.09±0.25
肝硬化组	53	27（50.8）^a	2.37±0.97	4.55±0.77	2.44±0.38
肝癌组	38	18（47.4）^a	4.23±1.08^b	5.53±1.18^b	4.74±0.57^b
正常对照组	20	0（0.0）	1.53±0.36	0	1.03±0.33

图2 乙肝肝癌患者血清中HBx蛋白表达与miR-143表达量的关系注：2A：HBx蛋白miR-143的相对表达量；2B：HBx蛋白水平与miR-143的关系

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[1]	汤永昌, 袁峰, 梁豪, 钟昭众, 熊志勇, 曹明波, 任昱朋, 李宇轩, 姚志成, 邓美海. HBx对HBV相关性肝癌增殖和迁移能力的影响及其机制[J]. 中华肝脏外科手术学电子杂志, 2022, 11(02): 198-202.

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Expression and clinical effect of the HBx protein mediated miR-143 in patients with hepatitis B hepatocellular carcinoma

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Expression and clinical effect of the HBx protein mediated miR-143 in patients with hepatitis B hepatocellular carcinoma