HMGB1对人肝癌细胞增殖及转移能力的影响

网络出版时间：2012-11-08 16:08

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HMGB1对人肝癌细胞增殖及转移能力的影响

刘亚萍，董蕾，史海涛，刘欣，鲁晓岚，姜炅

（西安交通大学医学院第二附属医院消化科，陕西西安710004)

摘要：目的观察高迁移率族蛋白1（HMGB1）对人肝癌细胞株HepG2体外增殖及转

移能力的影响，并阐明其可能的作用机制。方法MTT法检测细胞增殖能力及细胞对

Matrigel基质胶的粘附能力；Transwell小室模型测定细胞侵袭及迁移能力；Western blot

和逆转录PCR（RT-PCR）分别检测基质金属蛋白酶2(MMP-2)、基质金属蛋白酶

9(MMP-9)和血管内皮生长因子(VEGF)的蛋白及mRNA表达情况。结果HepG2经不

同质量浓度（10、100、1000ng/mL)的重组人HMGB1（rHMGB1）干预，24、48、72h

细胞增殖能力均明显高于对照组（P<0.01）；随着时间和浓度的增加，细胞增殖能力逐

渐增强。100ng/mL的rHMGB1干预HepG2 48h，细胞对Matrigel基质胶的粘附能力较

对照组明显增高(P<0.01)，侵袭和迁移实验中穿膜细胞数均明显多于对照组(P<0.01)。

此外，rHMGB1可明显上调MMP-9和VEGF的蛋白及mRNA表达水平(P<0.01)，但

MMP-2的蛋白及mRNA表达与对照组相比差异无统计学意义(P>0.05)。结论HMGB1

可促进人肝癌细胞的增殖、粘附、侵袭及迁移能力，这可能与上调MMP-9和VEGF

蛋白及mRNA表达水平有关。

关键词：HMGB1；肝癌细胞；增殖；肿瘤转移；基质金属蛋白酶2(MMP-2)；基质金

属蛋白酶9(MMP-9)；血管内皮生长因子(VEGF)

中图分类号：R735 文献标志码：A 文章编号：1671-8259

Effects of HMGB1 on proliferative and metastatic abilities of hepatoma cell line HepG2 LIU Ya-ping, DONG Lei, SHI Hai-tao, LIU Xin, LU Xiao-lan, JIANG Jiong

(Department of Gastroenterology, the Second Affiliated Hospital,

Medical School of Xi’an Jiao tong University, Xi'an 710004, China)

收稿日期：2012-05-20 修回日期：2012-09-16

基金项目：国家自然科学基金青年基金（No.81200310）

Supported by the Youth Fund of National Natural Science Foundation of China (No.81200310) 通讯作者：董蕾，主任医师，E-mail：dong556 @

作者简介：刘亚萍（1985-），女（汉族），硕士研究生，主要研究慢性肝病及胃肠动力.

E-mail: liuyaping5222491 @

ABSTRACT: Objective To investigate effects of high mobility group box-1 (HMGB1) on the proliferative and metastatic abilities of hepatoma cell line HepG2 in vitro and analyze the possible mechanisms. Methods MTT was used for determining the proliferative and adhesive abilities of HepG2. The invasive and migratory abilities were detected by Transwell assay. The expressions of MMP-2, MMP-9 and vascular endothelial growth factor (VEGF) at protein and mRNA levels were evaluated by Western blot and reverse transcription (RT-PCR), respectively. Results HepG2's proliferative ability in response to rHMGB1 of different concentration (10, 100 and 1000ng/mL) was significantly higher than that in the control group at 24, 48 and 72h (P<0.01), and it increased with the rise of treatment concentration and time. HepG2 was treated by rHMGB1 (100ng/mL) for 48 h to detect the adhesive, invasive and migratory abilities. Compared to that in the control group, the adhesive ability of HepG2 to Matrigel was significantly elevated (P<0.01) and the number of invasive and migratory cells was remarkably increased (P<0.01). In addition, the expressions of MMP-9 and VEGF at protein and mRNA levels were significantly up-regulated (P<0.01), but there was no difference in MMP-2 (P>0.05). Conclusion The proliferative, adhesive, invasive and migratory abilities of hepatoma cells can be promoted by HMGB1, which may be related to up-regulating the protein and mRNA expressions of MMP-9 and VEGF.

KEY WORDS: high mobility group box-1 (HMGB1); hepatoma cell; proliferation; tumor metastasis; MMP-2; MMP-9; vascular endothelial growth factor (VEGF)

高迁移率族蛋白1（high mobility group box-1, HMGB1）是真核细胞核内的非组蛋白染色质结合蛋白，在维持核小体结构的稳定和DNA重组、复制、修复及基因转录中发挥着重要作用[1]。HMGB1不仅作用于细胞内，还可以通过坏死细胞被动释放或免疫

[2]，其过表达可抑制细胞凋亡，诱导细胞增殖及分化[3]。新近研究表明HMGB1高表达于多种肿瘤组织[4]，参与肿瘤细胞的生长、浸润和迁移等生物学特性，具有重要的临床意义[5]。

原发性肝癌是临床上常见的恶性肿瘤之一，全球发病率逐年增长。其起病隐匿、恶性程度高，对于晚期肝癌患者来说手术效果欠佳、预后差，因此分子靶向治疗等生物治疗方法成为近年来研究的热点[6]，HMGB1作为一种新发现的肿瘤分子靶点，已有研究表明其在肝癌组织中有明显的表达[4]，但其对肝癌细胞的增殖及转移等生物学特性方面的影响尚不清楚。本研究即通过体外培养人肝癌细胞HepG2，观察HMGB1对其