摘要
为进一步探讨白细胞介素-2(IL-2)、可溶性白细胞介素-2受体(sIL-2R)、白细胞介素-4(IL-4)、碱性成纤维细胞生长因子(bFGF)、胰岛素样生长因子-Ⅱ(IGF-Ⅱ)及胰岛素样生长因子-Ⅱ受体(IGF-ⅡR)在肝癌发生、发展中的作用,为肝癌的早期诊断和应用细胞因子治疗肝癌提供实验依据,本研究以二乙基亚硝胺(DEN)诱发大鼠肝癌模型,应用酶联免疫吸附法(ELISA)、放射免疫测定法(RIA)及免疫组织化学(IHC)等方法,连续动态检测大鼠肝癌发生过程不同时段血清中IL-2、sIL-2R、IL-4、bFGF、IGF-Ⅱ的含量及IGF-Ⅱ及IGF-ⅡR在肝组织中的表达。结果显示:1.病理学检查证实DEN诱发的肝癌为肝细胞癌,诱癌率为70.6%(36/51),大鼠肝癌癌变过程大致经过肝细胞损伤期、肝细胞增生-硬化期和肝细胞癌变期等三个阶段,具有与人体肝癌相似的病变过程及病理学变化特征。2.大鼠血清IL-2水平随病变进展而逐渐降低,癌变组血清IL-2浓度低于正常对照组、肝细胞损伤组及增生-硬化组。3.诱癌过程中大鼠血清sIL-2R水平无明显变化。4.增生-硬化组大鼠血清IL-4水平明显低于正常对照组和肝细胞损伤组;癌变组血清IL-4浓度低于正常对照组、肝细胞损伤组及增生-硬化组。5.在诱癌过程中,大鼠肝组织及血清IGF-Ⅱ水平均呈“高→低→高”的变化趋势;IGF-Ⅱ及IGF-ⅡR在癌灶中的表达低于癌周增生灶、增生结节及非典型增生结节。6.大鼠血清bFGF水平的变化以癌变期为主,癌变组bFGF浓度显著低于正常对照组、肝细胞损伤组及增生-硬化组,bFGF与IGF-Ⅱ无明显相关。由此认为:1.饲以低剂量DEN诱发大鼠肝癌模型是一种较理想的研究人体肝癌发生的动物模型;2.检测血清IL-2水平有助于了解病情发展和机体免疫状态,是早期发现癌变的敏感指标;3.诱癌过程中大鼠血清IL-2及IL-4水平的变化未显示Th1/Th2漂移现象;4.IGF-Ⅱ与肝细胞增殖及恶性转化有关,但一旦肝细胞癌形成,IGF-Ⅱ可能就不再是重要的调节因子;5.诱癌初期大鼠血清IGF-Ⅱ水平的显著升高,提示IGF-Ⅱ可作为一个肝癌早期的诊断指标;6.诱癌过程中,bFGF与IGF-Ⅱ无协同作用。
In order to provide an experimental basis for early diagnosis and cytokine therapy on HCC (hepatocellular carcinoma), the expressions of IL-2, sIL-2R, IL-4, bFGF, IGF-Ⅱ, IGF-ⅡR during hepatocarcinogenesis in rats were studied. Experi- mental hepatocellular carcinomas were induced with the administration of diethylnitrosamine (DEN) in Wistar rats. The serum levels of IL-2, sIL-2R, IL-4 and bFGF were detected by enzyme linked immunosorbent assay (ELISA). Im- munohistochemical staining method was used for measuring the expressions of IGF-Ⅱand IGF-ⅡR in liver tissues in the carcinogenic successive stages. The serum level of IGF-Ⅱwas detected by radioimmunoassay (RIA). In this study, we found that: 1. 70.6% (36/51) hepatocellular carcinoma were induced by DEN. The procession of hepatocarcinogenesis in this model included three stages—hepatic toxic lesion, hepatic proliferation/cirrhosis and hepatic carcinogenesis. This ex- perimental hepatocarcinoma rat model induced by DEN is similar with the human hepatocellular carcinoma not only in forming procession but also in morphologic features. 2. The serum level of IL-2 was decreased in association with progression of HCC, it was higher in HCC group than that in control group, hepatic lesion group and proliferation/cirrhosis group. 3. There were no significant differences among all groups in the serum level of sIL-2R. 4. The serum level of IL-4 was significantly lower in proliferation/cirrhosis group than that in control group and hepatic lesion group, and it was lower in HCC group than those in other groups. 5. The IGF-Ⅱpresent an pattern of "high-low-high" in terms of degree of expression not only in liver tissues but also in serum during carcinogenesis.The expressions of IGF-Ⅱand IGF-ⅡR in HCC were lower than those in the paratumor foci, para- tumor hyperplastic nodules and paratumor atypical hyperplastic nodules. 6. The serum level of bFGF changed mainly in the stage of hepatic carcinogenesis, it was lower in HCC group than those in other groups. There was no correlation between bFGF and IGF-Ⅱ. It may be concluded that: 1. The liver cancer model induced by low dose DEN is an ideal experimental model for the study of hepatocarcino- genesis. 2. Measuring the serum level of IL-2 can monitor the pathogenetic condition and systematic immunologic status. The serum level of IL-2 is a sensitive index to early hepatocarcinogenesis. 3. Th1/Th2 cytokine imbalance is not observed. 4. IGF-Ⅱmight be associated with hepatocytic proliferation and malignant transformation of hepatocytes. Once tumours formed, probably it was not an important regulatory factor any longer. 5. The essential role of IGF-Ⅱat the immediate initiation stage of carcinogenesis can not be ignored. Thus this expression can be used as a suitable marker for early detection of the cancerous process. 6. The two angiogenic growth factors, bFGF and IGF-Ⅱ, have no synergistic effect in the carcinogenesis and progression of HCC.
引文
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