家族聚集性肝细胞肝癌组织中S期激酶相关蛋白2和增殖细胞核抗原的表达及意义

doi:10.3877/cma.j.issn.2095-3232.2013.04.012

目的

探讨S期激酶相关蛋白2（Skp2）、增殖细胞核抗原（PCNA）在家族聚集性肝细胞肝癌（肝癌）组织中的表达及意义。

方法

回顾性研究2001年1月至2010年1月在中山大学附属第三医院行肝癌肝切除的79例患者临床资料。所有患者均签署知情同意书，符合医学伦理学规定。根据是否符合家族聚集性肝癌的诊断分为家族聚集性肝癌组（FH组)和非家族聚集性肝癌组（NFH组）。FH组34例，其中男6例，女28例；年龄（34±5）岁。NFH组45例，其中男8例，女37例；年龄（59±4）岁。另取40例正常肝组织作为对照。应用免疫组织化学方法（免疫组化法）检测Skp2、PCNA的表达，蛋白质印迹法检测Skp2、PCNA的蛋白含量。一般资料中的计量资料和Skp2、PCNA的蛋白含量比较采用t检验，一般资料中的定性资料和Skp2、PCNA的阳性表达比较采用χ²检验。

结果

FH组与NFH组患者的性别、年龄、乙型肝炎表面抗原（HBsAg）阳性率、乙型肝炎病毒脱氧核糖核酸（HBV-DNA）定量、甲胎蛋白（AFP）、肝功能Child-Pugh评分、肿瘤有否肝外转移、有否肿瘤包膜、有否静脉癌栓等临床资料的比较差异均无统计学意义（P>0.05）。FH组肝癌组织Skp2、PCNA阳性表达的数密度分别为（37±3）、（57±5）个/高倍镜视野，NFH组分别为（17±3）、（38±4）个/高倍镜视野，FH组肝癌组织中的Skp2、PCNA阳性表达明显高于NFH组（t=2.12，3.14；P<0.05)。蛋白质印迹法检测结果发现，FH组肝癌组织中Skp2、PCNA的蛋白含量分别为2.7±0.3、2.4±0.4，NFH组分别为1.7±0.4、1.5±0.3，FH组肝癌组织中Skp2、PCNA的蛋白含量明显高于NFH组（t=0.15，0.13；P<0.05)。

结论

肝癌组织中Skp2、PCNA蛋白阳性表达上调，家族聚集性肝癌患者的肝癌组织中Skp2、PCNA的表达明显强于非家族聚集性肝癌患者的肝癌组织。

关键词: 癌，肝细胞, 家族聚集性肝癌, S期激酶相关蛋白类, 增殖细胞核抗原, 免疫组织化学, 印迹法，蛋白质

Objective

To investigate the expression and significance of S-phase kinase associated protein 2(Skp2) and proliferating cell nuclear antigen(PCNA) in the tissues of familial aggregation hepatocellular carcinoma (HCC).

Methods

Clinical data of 79 HCC patients who received resection of liver cancer from January 2001 to January 2010 in the Third Affiliated Hospital of Sun Yat-Sen University were analyzed retrospectively. The informed consents of all patients were obtained and the ethical committee approval was received. The patients were divided into two groups: familial aggregation HCC group [FH group; 6 males and 28 females; mean age of (34±5) year old] and non-familial aggregation HCC group [NFH group; 8 males and 37 females; mean age of (59±4) year old]. Another 40 cases with normal liver tissues were selected as control. The expression of Skp2 and PCNA were detected by immunohistochemistry and the protein contents of Skp2 and PCNA were detected by Western blot. Measurement data of the general clinical data and protein contents of Skp2 and PCNA were compared by t test. Qualitative data of the general clinical data and positive expression of Skp2 and PCNA were compared by Chi-square test.

Results

There was no significant difference between FH group and NFH group in sex, age, positive rate of hepatitis B surface antigen(HBsAg), quantitation of hepatitis B virus-deoxyribonucleic acid(HBV-DNA), the level of alpha fetal protein(AFP), Child-Pugh score, presence or absent of extrahepatic metastasis of tumor, tumor capsule and venous tumor emboli(P>0.05). Compared with NFH group, the number densities of Skp2 and PCNA positive expression in FH group were significantly higher[(17±3)/high power field vs. (37±3)/high power field, (38±4)/high power field vs. (57±5)/high power field; t=2.12, 3.14; P<0.05]. The results of Western blot assay showed that the protein contents of Skp2 and PCNA of HCC tissues in FH group were significantly higher than those in NFH group[(2.7±0.3) vs. (1.7±0.4), (2.4±0.4) vs. (1.5±0.3); t=0.15, 0.13; P<0.05].

Conclusions

The positive expression of Skp2 and PCNA increase in the tissues of HCC. The expression of Skp2 and PCNA in the tissues of familial aggregation HCC obviously increase than that in the tissues of non-familial aggregation HCC.

Key words: Carcinoma, hepatocellular, Familial aggregation hepatocellular carcinoma, S-phase kinase-associated proteins, Proliferating cell nuclear antigen, Immunohistochemistry, Blotting, western

表1 两组肝癌患者计数资料的比较（例）

组别	例数	性别		HBsAg		肝外转移		肿瘤包膜		静脉癌栓
组别	例数	男	女	阳性	阴性	无	有	无	有	无	有
FH组	34	6	28	34	0	29	5	23	11	29	5
NFH组	45	8	37	32	13	38	7	35	10	38	7
χ²值	?	0.623		0.929		0.635		0.912		0.798
P值	?	0.751		0.124		0.621		0.364		0.387

表1 两组肝癌患者计数资料的比较（例）

组别	例数	性别		HBsAg		肝外转移		肿瘤包膜		静脉癌栓
组别	例数	男	女	阳性	阴性	无	有	无	有	无	有
FH组	34	6	28	34	0	29	5	23	11	29	5
NFH组	45	8	37	32	13	38	7	35	10	38	7
χ²值	?	0.623		0.929		0.635		0.912		0.798
P值	?	0.751		0.124		0.621		0.364		0.387

表2 两组肝癌患者计量资料的比较（±s）

组别	例数	年龄（岁）	HBV-DNA(IU/ml)	AFP(μg/L)	Child-Pugh评分（分）
FH组	34	34±5	751±70	231±20	5.1±2.1
NFH组	45	59±4	351±51	271±31	4.9±3.2
t值	?	0.674	0.732	0.413	0.876
P值	?	0.112	0.131	0.751	0.754

表2 两组肝癌患者计量资料的比较（±s）

组别	例数	年龄（岁）	HBV-DNA(IU/ml)	AFP(μg/L)	Child-Pugh评分（分）
FH组	34	34±5	751±70	231±20	5.1±2.1
NFH组	45	59±4	351±51	271±31	4.9±3.2
t值	?	0.674	0.732	0.413	0.876
P值	?	0.112	0.131	0.751	0.754

图1 两组肝细胞肝癌患者癌组织Skp2免疫组织化学方法染色结果（×400）

图2 两组肝细胞肝癌患者癌组织的PCNA免疫组织化学方法染色结果（×200）

图3 两组肝细胞肝癌患者癌组织Skp2蛋白质印迹电泳图

图4 两组肝细胞肝癌患者癌组织PCNA蛋白质印迹电泳图

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Expression and significance of S-phase kinase associated protein 2 and proliferating cell nuclear anti-gen in the tissues of familial aggregation hepatocellular carcinoma

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Expression and significance of S-phase kinase associated protein 2 and proliferating cell nuclear anti-gen in the tissues of familial aggregation hepatocellular carcinoma