HMGB1对宫颈癌干性基因OCT4、Sox2和Nanog表达的影响

Effect of High-mobility Group Box-1 on Expression of Stem Cell Markers OCT4, Sox2 and Nanog in Cervical Carcinoma

目的研究高迁移率族蛋白B1(HMGB1)对宫颈癌干性基因OCT4、Sox2和Nanog的作用。方法构建HMGB1过表达质粒及干扰HMGB1表达的特异性siRNA,将上述重组质粒和(或)siRNA转染到宫颈癌细胞中。采用Western blot和流式细胞术检测HMGB1上调(或抑制)表达后对上述干细胞标志物表达的影响。免疫组织化学法检测HMGB1、OCT4、Sox2和Nanog在66例宫颈组织中的表达并分析相关性。结果 (1)结果显示HMGB1编码区全长为648 bp,其与GV230真核表达系统构建出HMGB1过表达载体,同时筛选出干扰效率的最高的siRNA-HMGB1-1来进行后续实验;(2)与空载体组和HMGB1低表达组相比,HMGB1过表达组细胞株中干细胞标志物OCT4、Sox2和Nanog蛋白表达和比例明显增高,差异具有统计学意义(均P<0.05);HMGB1低表达时,Nanog、OCT4和Sox2蛋白的表达和比例明显降低,差异具有统计学意义(均P<0.05);(3)在宫颈鳞癌组织中HMGB1、OCT4、Sox2和Nanog蛋白阳性表达明显高于慢性宫颈炎组,HMGB1的表达与干细胞标志物的表达呈正相关。结论 HMGB1能够调节宫颈癌干性基因OCT4、Sox2和Nanog的表达,可能是抗肿瘤治疗的潜在靶点。

Objective To investigate the role of High-mobility group box-1（HMGB1） protein in regulating the expression of cancer stem cell markers Nanog, OCT4 and Sox2. Methods We transferred recombinant plasmid and target-specific siRNA of HMGB1 into cervical cancer cells. The expression of cancer stem cell markers（OCT4, Sox2 and Nanog） were measured by Western blot and flow cytometry. Immunohistochemical method was applied to detect HMGB1, OCT4, Sox2 and Nanog expression in 66 cases of cervical tissues. Rank correlation analysis was used to analyze their correlation. Results The full-length coding sequence of HMGB1 was 648 bp and recombined to overexpress the plasmid GV230. The selection of the highest interference rate was carried out for subsequent experiments. The protein expression and rates of stem cell markers OCT4, Sox2 and Nanog were significantly higher in HMGB1 overexpression group than those in low HMGB1 expression group and empty vector group（P〈0.05）. The protein expression and rates of stem cell markers OCT4, Sox2 and Nanog were significantly lower in low HMGB1 expression group than those in HMGB1 overexpression group and empty vector group（P〈0.05）. HMGB1, OCT4, Sox2 and Nanog protein expression in cervical carcinoma tissues were higher than those in chronic cervicitis tissues. Positive correlations were also found between HMGB1 and OCT4, Sox2 and Nanog expression. Conclusion HMGB1 could regulate the expression of cancer stem cell markers Nanog, OCT4 and Sox2, therefore, it is probably a potential target for anti-tumor therapy.