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[目的]乙型肝炎病毒(Hepatitis B Virus,HBV)在全球广泛流行,是引起乙型肝炎(Viral Hepatitis)和原发性肝细胞癌(Primary Hepatocellular Carcinoma,PHCC)的主要原因之一。我们采用根据HBV-X基因的序列设计针对起始密码的脱氧核酶来阻断HBV-X基因的表达,探寻X基因与肝细胞癌发生的机制,研究防治肝细胞癌的方法,对临床上进行肝癌基因治疗提供理论依据。[方法]应用Western blot、流式细胞仪以及RT-PCR等检测脱氧核酶对X基因的抑制作用。[结果]根据HBV-X基因的序列设计针对起始密码的脱氧核酶,导入能表达HBV-X基因的细胞系,通过流式细胞仪、Western blot以及RT-PCR等检测脱氧核酶在细胞系中对HBV-X基因的作用情况,发现DNA核酶对HBV-X基因在肝癌细胞中的表达有一定抑制作用。[结论]利用脱氧核酶来阻断乙肝病毒X基因表达从而防治肝细胞癌的发生,降低肝细胞癌的死亡率,提高慢性HBV感染者的生存质量,具有很大的社会效益及经济效益。
[Objective] Hepatitis B virus (HBV) is widely prevalent in the world and is one of the main causes of Viral Hepatitis and Primary Hepatocellular Carcinoma (PHCC). We designed a DNAzyme-based deoxyribozyme based on the sequence of HBV-X gene to block the expression of HBV-X gene, explore the mechanism of X gene and hepatocellular carcinoma, and study the method of prevention and treatment of hepatocellular carcinoma. Provide a theoretical basis for liver cancer gene therapy. [Method] The inhibitory effect of DNAzyme on X gene was detected by Western blot, flow cytometry and RT-PCR. [Results] According to the sequence of HBV-X gene, a deoxyribozyme targeting the initiation codon was designed and introduced into a cell line expressing HBV-X gene. Flow cytometry, Western blot and RT- Department of HBV-X gene in the role of the situation and found that DNAzyme on HBV-X gene in liver cancer cells have a certain degree of inhibition. [Conclusion] The use of DNAzyme to block the expression of hepatitis B virus X gene in order to prevent and treat the occurrence of hepatocellular carcinoma, reduce the mortality of hepatocellular carcinoma and improve the quality of life of patients with chronic HBV infection has great social and economic benefits.