肝星状细胞对肝癌SMMC-7721细胞增殖、侵袭的影响及作用机制研究

Effect of hepatic stellate cells on proliferation and invasiveness of human hepatocellular carcinoma cells

目的 观察人肝星状细胞LX-2对人肝癌SMMC-7721细胞增殖、侵袭能力的影响,并探讨其可能的作用机制。方法 人肝星状细胞LX-2条件培养基(HSC-CM)与肝癌SMMC-7721细胞共培养,设为HSC-CM组;人正常肝细胞LO2条件培养基(LO2-CM)与肝癌SMMC-7721细胞共培养,设为LO2-CM组(阴性对照);1%FBS培养基与肝癌SMMC-7721细胞共培养,设为1%FBS组(空白对照)。采用CCK-8法检测3组肝癌SMMC-7721细胞增殖能力;Transwell侵袭实验检测培养48h后细胞侵袭能力;Western blot检测增殖细胞核抗原(PCNA)、血管内皮生长因子(VEGF)、核因子κB(NF-κB)、基质金属蛋白酶2(MMP-2)蛋白表达水平。结果 HSC-CM组、LO2-CM组、1%FBS组肝癌SMMC-7721细胞增殖能力、侵袭能力、PCNA、VEGF、NF-κB、基质金属蛋白酶2(MMP-2)蛋白表达水平比较差异均有统计学意义(均P<0.05);与LO2-CM组、1%FBS组比较,HSC-CM组肝癌SMMC-7721细胞增殖能力、侵袭能力均增强(均P<0.05);PCNA、VEGF、NF-κB、MMP-2蛋白表达水平均上调(均P<0.05)。结论 肝星状细胞可促进肝癌SMMC-7721细胞增殖、侵袭等生物学行为,其作用机制可能与上调PCNA、VEGF、NF-κB、MMP-2表达水平有关。

Objective To investigate the effect of human hepatic stellate cell LX-2 on the proliferation and invasiveness of human hepatocellular carcinoma cells, and its mechanism. Methods Human hepatic stellate LX-2 cells in conditioned medium（HSC-CM） was co-cultured with human hepatocellular carcinoma SMMC-7721 cells（HSC-CM group）; human normal liver LO2 cells in conditioned medium（LO2-CM） were co-cultured with SMMC-7721 cells（negative control）; hepatoma SMMC-7721 cells were cultured in 1% FBS medium（blank control）. CCK-8 assay was used to detect the proliferation of SMMC-7721 cells in three groups. Transwell assay was used to detect the cell invasion ability after 48 h of culture. Proliferating cell nuclear antigen（PCNA）, vascular endothelial growth factor（VEGF） and nuclear factor-κB（NF-κB）, matrix metalloproteinase 2（MMP-2） protein expression levels were detected by Western blot. Results Compared with LO2-CM group and 1% FBS group, the proliferation and invasion of HSC-CM group were all enhanced（all P〈0.05）; protein expression levels of PCNA, VEGF, NF-κB, MMP-2 were up-regulated（all P〈0.05）. Conclusion Hepatic stellate cells can promote the proliferation and invasion of hepatocellular carcinoma SMMC-7721 cells, which are associated with the up-regulation of PCNA, VEGF, NF-κB, MMP-2 expression.