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目的利用树鼩cDNA芯片研究黄曲霉毒素B1(AFB1)和(或)乙型肝炎病毒(HBV)引起的树鼩肝细胞癌(HCC)发生过程中参与信号传导的部分基因表达变化情况,从而进一步探讨HCC发生的分子机制。方法实验树鼩分3组:A组(AFB1组)、B组(AFB1+HBV组)、C组(正常对照组)。所有动物在实验过程中定期接受剖腹手术取肝组织检查,至肝癌形成时处死动物取肝癌和癌旁组织。用树鼩CD N A 芯片检测实验第30、60、90周肝活检组织、肝癌组织及其癌旁组织中各基因的表达情况,并用实时逆转录聚合酶链反应(Real time RT-PCR)法验证cDNA芯片结果。结果在A组和B组从癌前到癌变过程中胰岛素样生长因子-Ⅱ(IGF-Ⅱ)、C-rel、核因子-κB2(NF-κB2)均显示有差异表达, 同时bcl-2、细胞周期素A(cyclin A)、睫状神经营养因子(cNTF)仅在B组显示有差异表达。而C组这几个基因无差异表达。实验组与对照组比较,在诱癌的早、中期(30、60周)均出现CNTF及cyclin A的差异表达。real time RT PCR结果与cDNA芯片检测结果基本一致。A组IGF-Ⅱ、C-rel基因及B组IGF-Ⅱ基因,在肝癌组织表达水平明显低于癌旁及实验30、60周组织,而癌旁与实验30、60周相比较则无明显差异;B组CNTF 基因在癌旁,肝癌及60周之间比较无明显改变,但均明显高于30周;A组CNTF基因表达水平在癌旁、肝癌组织高于癌前组织,但差异无统计学意义,C组这3个基因不同时期比较亦无明显差异。结论树鼩部分信号传导通路相关cDNA芯片应用于检测树鼩HCC发生过程中信号传导通路中各基因表达的变化,对进一步了解树鼩HCC发生的机制有重要实用价值。IGF-Ⅱ、NF-κB2、C-rel、Bcl-2、cyclin A及CNTF 这几个基因与树HCC的发生发展密切相关。
Objective To study the changes of some genes involved in signal transduction in tree shrew hepatocellular carcinoma (HCC) caused by aflatoxin B1 (AFB1) and / or hepatitis B virus (HBV) by cDNA library of tree shrews To investigate the molecular mechanism of HCC. Methods The experimental tree was divided into three groups: group A (AFB1), group B (AFB1 + HBV) and group C (normal control). All animals underwent routine laparotomy in the course of the experiment to take the liver tissue examination, and the animals were sacrificed at the time of liver cancer formation to take liver cancer and paracancer tissues. The expression of each gene in the liver biopsy, liver cancer tissues and their adjacent tissues at the 30th, 60th, 90th week of the experiment was detected by the CDNA chip of tree 鼩 and verified by real time RT-PCR cDNA chip results. Results The expressions of IGF-Ⅱ, C-rel and NF-κB2 were significantly different in group A and group B from precancerous to carcinogenesis, while the expressions of bcl-2, Cyclin A and cNTF showed differential expression in group B only. The C group of these genes no difference in expression. Compared with the control group, the difference of CNTF and cyclin A in the early and mid-term (30 and 60 weeks) of induction of cancer were observed. Real time RT PCR results and cDNA chip test results are basically the same. The expression of IGF-Ⅱ and C-rel in group A and IGF-Ⅱ in group B were significantly lower than that in the paracancer and the tissues at 30 and 60 weeks, while there was no significant difference between paracancer and 30 and 60 weeks ; The expression of CNTF in group B was significantly higher than that in group of adjacent tissues and liver cancer for 60 weeks, but both were significantly higher than those in group A; the expression of CNTF in group A was higher than that in precancerous tissues and hepatocellular carcinoma tissues, but the difference was not statistically significant Significance of learning, C group of these three genes in different periods also showed no significant difference. Conclusion The cDNA microarray associated with part of signal transduction pathways in tree shrews is useful for detecting the expression of various genes in signal transduction pathways during the occurrence of tree shrew HCC. It is of great practical value to further understand the mechanism of tree shrew HCC. The genes IGF-Ⅱ, NF-κB2, C-rel, Bcl-2, cyclin A and CNTF are closely related to the occurrence and development of HCC.