HBV相关肝细胞癌组织中CDC37基因启动子微卫星多态性及其与基因表达的关系

Correlation between microsatellite polymorphism in promoter of CDC37 and its expression in tissues of HBV-related HCC

目的：探讨乙型肝炎病毒（ HBV）相关肝细胞癌（ HCC）组织中CDC37基因启动子区微卫星的多态性及其与基因表达的关系。方法22例接受手术治疗的HBV相关HCC患者，分别提取其癌组织和癌旁组织中的基因组DNA，PCR扩增CDC37基因启动子区含微卫星区段序列，并克隆至PMD-18T载体，每份标本至少挑选3个克隆进行测序，然后对HCC组织和癌旁组织、CDC37过表达的HCC组织与相应的癌旁组织、CDC37过表达和无过表达的HCC组织的启动子GT微卫星多态性组成形式进行比较分析。结果本组患者HCC组织和癌旁组织中CDC37基因启动子区存在（GT）n1-GC-（GT）n2多种类型的微卫星形式，位于CDC37启动子区1368～1415位，以（GT）10-GC-（GT）9最为常见。 HCC组织与癌旁组织的CDC37启动子微卫星多态性形式的总体分布相比，P＞0．05；CDC37过表达HCC组织与相应的癌旁组织的微卫星的总体分布相比，P＞0．05；而CDC37过表达和无过表达HCC组织的微卫星的总体分布相比，P＜0．05，其中（GT）10-GC-（GT）9形式在CDC37过表达患者中所占比例高于无过表达患者，两者相比，P＜0．05，（GT）10-GC-（GT）12形式在CDC37无过表达患者中所占比例高于过表达患者，两者相比，P＜0．05。结论 HBV相关HCC患者癌组织中CDC37基因启动子区存在微卫星多态性，CDC37过表达和无过表达HCC组织的微卫星形式有不同。

Objective To explore the relationship between the microsatellite polymorphism in the promoter of CDC 37 and its expression in tissues of HBV-related HCC.Methods Genomic DNA was isolated from tissues of tumor and non-tumor of 22 patients with HBV-related HCC.Then the DNA was amplified using PCR to span the microsatellite of the hu-man CDC37 promoter.PCR products were cloned into PMD-18T vector.At least 3 clones of each specimen were se-quenced.The types and frequencies of（GT）n repeats were compared between：tumor and non-tumor of all patients;tumor and non-tumor of patients with CDC37 over-expression; tumor with and without CDC37 over-expression.Results Many types of microsatellite（GT）n1-GC-（GT）n2 were found in CDC37 gene promotor in tumor and non-tumor of the patients.The microsatellite was located at nt 1 368 to nt 1 415 of CDC37 promoter.The（GT）10-GC-（GT）9 was the most prevalent type. The distribution of the types of （ GT ） n repeats showed no difference between tumor and non-tumor of all patients （ P 〉0.05）.No statistical difference was also found between tumor and non-tumor of patients with CDC37 over-expression（P〉0.05）.Significant difference was found between tumor with and without CDC 37 over-expression（P〈0.05）.The （GT）10-GC-（ GT） 9 repeats showed a higher frequency in tumors with CDC 37 over-expression than in tumors without CDC 37 over-ex-pression（P〈0.05）, while the （GT）10-GC-（GT）12 repeats showed a higher prevalence in tumors without CDC 37 over-ex-pression comparing with tumors with CDC37 over-expression（P〈0.05）.Conclusions Microsatellite polymorphism is found in the promoter of CDC37 gene in tissues of tumor and non-tumor of patients with HBV-related HCC.The distribution of the types of （GT）n repeats shows statistical difference between tumor with and without CDC 37 over-expression.