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基质胶的黏附能力;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、1 000ng/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基质胶的黏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、黏附、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。
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 to determine the proliferative and adhesive abilities of HepG2. The invasive and migratory abilities were detected by Transwell assay. The protein and mRNA expressions of matrix metalloproteinases-2 (MMP-2), matrix metalloproteinases-9(MMP-9) and vascular endothelial growth factor (VEGF) were evaluated by Western blot and reverse transcription (RT-PCR), respectively. Results The proliferative ability of HepG2 in response to rHMGB1 of different concentrations (10, 100 and 1 000 ng/mL) was significantly higher than that in the control group at 24, 48 and 72 h (P〈0.01), and it increased with the increase of treatment concentration and time. HepG2 was treated by rHMGB1 (100 ng/mL) for 48 h to detect the adhesive, invasive and migratory abilities. Compared to 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 protein and mRNA expressions of MMP-9 and VEGF were significantly up-regulated (P〉0.01), but there was no difference in the protein and mRNA expressions of 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.
出处
《西安交通大学学报(医学版)》
CAS
CSCD
北大核心
2013年第2期159-163,共5页
Journal of Xi’an Jiaotong University(Medical Sciences)
基金
国家自然科学基金青年基金资助项目(No.81200310)~~