Li Xiangnan, Zhu Fangyu, He Yongsong, Luo Fang
Department of Hepatobiliary Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Department of Hepatobiliary Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Apr;33(4):477-482.
Objective To investigate the cell inhibitory effect of arginase inhibitor nor-NOHA on HepG2 hepatocellular carcinoma cells and related mechanism. Methods CCK-8 assay was used to detect the cell proliferation and flow cytometry to detect the apoptosis of HepG2 cells treated with (0, 0.5, 1.0, 2.0, 3.0) ng/μL nor-NOHA. The protein levels of arginase 1 (Arg1), P53, matrix metalloproteinase-2 (MMP-2), E-cadherin (ECD) were determined by Western blotting. Real time quantitative PCR was employed to examine the changes in the mRNA level of inducible nitric oxide synthase (iNOS). Griess assay was used to measure the concentration of nitric oxide (NO) in HepG2 cells. Transwell assay and wound-healing assay were performed to evaluate the changes of the cell invasion and migration ability, respectively. Results nor-NOHA inhibited the proliferation and induced the apoptosis of HepG2 cells. It also decreased the expression levels of Arg1 and MMP-2, increased the expression levels of P53 and ECD as well as the production of NO; in addition, nor-NOHA inhibited the invasion and migration of HepG2 cells. Conclusion Nor-NOHA can induce cell apoptosis and inhibit the ability of invasion and migration of HepG2 cells by inhibiting Arg1, which is related with the increase of iNOS expression and the high concentration of NO.
目的 探讨精氨酸酶抑制剂nor-NOHA对HepG2肝癌细胞的抑制作用及相关机制。方法 采用CCK-8法检测(0、0.5、1.0、2.0、3.0)ng/μL的nor-NOHA处理HepG2细胞后的增殖情况,流式细胞术检测细胞凋亡情况。采用蛋白质印迹法检测精氨酸酶1(Arg1)、P53、基质金属蛋白酶-2(MMP-2)、E-钙黏蛋白(ECD)的蛋白水平。采用实时定量PCR检测诱导型一氧化氮合酶(iNOS)mRNA水平的变化。采用Griess法检测HepG2细胞中一氧化氮(NO)的浓度。分别采用Transwell实验和划痕实验评估细胞侵袭和迁移能力的变化。结果 nor-NOHA抑制HepG2细胞的增殖并诱导其凋亡,降低Arg1和MMP-2的表达水平,增加P53和ECD的表达水平以及NO的生成;此外,nor-NOHA抑制HepG2细胞的侵袭和迁移。结论 Nor-NOHA可通过抑制Arg1诱导细胞凋亡并抑制HepG2细胞的侵袭和迁移能力,这与iNOS表达增加和NO浓度升高有关。