Department of Medical Oncology, The Fifth People's Hospital of Changshu, Changshu, China.
Department of Gastroenterology, JingJiang People's Hospital, Jingjiang, China.
Neoplasma. 2020 Jan;67(1):61-67. doi: 10.4149/neo_2019_190221N147. Epub 2019 Nov 26.
The present study aimed to assess the association of rs34000982 polymorphism located in the 3'untranslated region (3'UTR) of high mobility group box 1 (HMGB1) gene and the risk of hepatocellular carcinoma (HCC), and further to explore the underlying mechanism. Genomic DNA was extracted from peripheral blood of 320 patients with HCC and 360 matched controls. Rs34000982 polymorphism was genotyped by a polymerase chain reaction-polyacrylamide gel electrophoresis assay. The genotype-phenotype association of HMGB1 mRNA and protein expression in HCC tissues with different genotypes was detected by quantitative (q) PCR assay and western blot. Vectors containing the insertion (ins)/ins or deletion (del)/del genotype of the rs34000982 polymorphism were constructed and the HMGB1 transcriptional activity affected by the rs34000982 polymorphism was detected by the luciferase assay. It was identified that the ins/ins genotype of rs34000982 significantly increased the risk of HCC compared with the del/del genotype. Further the qPCR results demonstrated that the HMGB1 mRNA expression level in HCC tissues with ins/ins genotype was 2.24 times that of HCC tissues with ins/del and del/del genotypes and there was a similar trend at protein level. In addition, the insertion allele of rs34000982 disturbed the binding of miR-636 with the 3'UTR of HMGB1, thereby increasing HMGB1 transcriptional activity in vitro. These data suggest that the rs34000982 polymorphism may contribute to HCC susceptibility, in full or at least partially through the effect on HMGB1 transcriptional activity by disturbing the binding of miR-636 with the 3'UTR of HMGB1.
本研究旨在评估位于高迁移率族蛋白 B1 (HMGB1) 基因 3'非翻译区（3'UTR）的 rs34000982 多态性与肝细胞癌（HCC）风险的关联，并进一步探讨其潜在机制。从 320 例 HCC 患者和 360 例匹配对照者的外周血中提取基因组 DNA。采用聚合酶链反应-聚丙烯酰胺凝胶电泳法检测 rs34000982 多态性。采用 qPCR 法和 Western blot 法检测 HCC 组织中不同基因型 HMGB1 mRNA 和蛋白表达的表型-基因型关联。构建包含 rs34000982 插入（ins）/ins 或缺失（del）/del 基因型的载体，并通过荧光素酶检测 rs34000982 多态性对 HMGB1 转录活性的影响。结果发现，与 del/del 基因型相比，rs34000982 的 ins/ins 基因型显著增加 HCC 的发病风险。进一步的 qPCR 结果表明，ins/ins 基因型 HCC 组织中 HMGB1 mRNA 表达水平是 ins/del 和 del/del 基因型 HCC 组织的 2.24 倍，蛋白水平也有类似趋势。此外，rs34000982 的插入等位基因干扰了 miR-636 与 HMGB1 3'UTR 的结合，从而增加了体外 HMGB1 的转录活性。这些数据表明，rs34000982 多态性可能导致 HCC 易感性增加，其作用可能至少部分是通过干扰 miR-636 与 HMGB1 3'UTR 的结合来影响 HMGB1 的转录活性。
