高迁移率族蛋白B1与肝癌细胞再增殖及肝癌患者预后的关系

Association of high mobility group box 1 with liver cancer cell repopulation and prognosis of liver cancer patients

背景与目的:肝癌是消化系统常见恶性肿瘤,肿瘤复发是导致其治疗失败的主要原因。高迁移率族蛋白B1 (HMGB1)被证实在多种肿瘤组织中高表达,在肿瘤发生发展中扮演着重要角色。本研究通过肝癌细胞再增殖体外模型与数据库分析,探讨HMGB1与肝癌细胞再增殖以及与肝癌预后的关系。方法:选择肝癌细胞Huh7和Li7作为研究对象,并通过转染萤火虫荧光素蛋白-水母绿色荧光蛋白融合蛋白(Fluc-GFP)基因质粒构建相对应的报告细胞Huh7-Fluc和Li7-Fluc。以10 Gy剂量X射线照射过的Huh7和Li7作为饲养细胞,分别与相应的报告细胞Huh7-Fluc和Li7-Fluc共培养构建肿瘤再增殖模型,以单纯的报告细胞以及报告细胞与无X射线处理的饲养细胞共培养的体系为对照,通过生物成像观察荧光素酶活性变化来判断报告细胞生长情况,以及HMGB1抑制剂甘草酸(Gly)干预后的影响。通过TIMER2.0和GEPIA2数据库分析平台,分析HMGB1在肝癌组织及癌旁组织的表达水平以及与肝癌预后的相关性。结果:与各自单纯报告细胞培养组以及报告细胞加无X射线处理饲养细胞共培养组比较,再增殖模型组报告细胞增殖能力明显增强(均P<0.01)。Gly干预后,再增殖模型组报告细胞的生长被明显抑制(均P<0.01),但单纯培养的报告细胞的生长无明显影响(均P>0.05)。公共数据库检索结果显示,HMGB1在肝癌组织中的表达水平明显高于癌旁组织(P<0.01),HMGB1高表达的肝癌患者总生存期短(P<0.01),HMGB1的表达水平与肝癌患者的无病生存期无相关性(P>0.05)。结论:HMGB1参与了X射线诱导的肝癌细胞再增殖。HMGB1在肝癌组织中的表达水平,可作为肝癌患者的总生存期预后评估参考指标,并为肝癌的临床治疗提供新思路。

Background and Aims: Liver cancer is a common malignant tumor of the digestive system. Tumor recurrence is a major cause of treatment failure. High mobility group box 1(HMGB1) has been found to be highly expressed in a variety of cancers, and play an important role in cancer development and progression. Therefore, this study was designed to investigate the relations of HMGB1 with the repopulation of liver cancer cells and the prognosis of liver cancer through in vitro repopulation model of liver cancer cells and database analysis.Methods: Liver cancer Huh7 and Li7 cells were chosen for this study. The two types of cells were transfected with plasmids carrying firefly luciferin and green fluorescent protein(Fluc-GFP) gene to construct respective reporter cells Huh7-Fluc and Li7-Fluc cells. The Huh7 and Li7 cells treated with Xray irradiation at a dose of 10 Gy were used as feeder cells and then co-cultured with their corresponding reporter cells to construct the in vitro repopulation models, using the systems of pure reporter cells and co-culture of the reporter cells and feeder cells without X-ray treatment as controls. The growth conditions of the reporter cells were evaluated by analyzing the changes in their luciferase activities via bioimaging, on which the effects of HMGB1 inhibitor glycyrrhizin(Gly) intervention were also observed. The HMGB1 expression levels of liver cancer tissue and tumor adjacent tissue as well as its association with the prognosis of liver cancer were analyzed using the TIMER2.0 and GEPIA2 tumor data analysis platforms.Results: Compared with respective pure reporter cells group and reporter cells plus non-irradiated feeder cells co-culture group, the growth abilities of the reporter cells in both repopulation groups were significantly enhanced(all P<0.01). After Gly intervention, the growth abilities of the reporter cells in both repopulation models were significantly suppressed(both P<0.01), but the growth conditions of the pure reporter cells showed no significant changes(both P>0.05). The results of public database retrieval showed that the expression level of HMGB1 was higher in liver cancer than that in adjacent tissue(P<0.01), and the overall survival time of liver cancer patients with high HMGB1 expression level was significantly shorter than those with low HMGB1 expression level(P<0.01), and HMGB1 expression level was not correlated with disease free survival of liver cancer patients(P>0.05).Conclusion: HMGB1 is involved in liver cancer cell repopulation induced by X-ray irradiation. The HMGB1 expression level in liver cancer tissue can be used as reference index for predicting the prognosis of overall survival of liver cancer patients, which may also provide a new strategy for the clinical treatment of liver cancer.