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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其机制。方法采用CCK-8法及流式细胞术检测(0.0、0.5、1.0、2.0、3.0)ng/μL nor-NOHA对人肝癌HepG2细胞的增殖抑制作用和细胞凋亡的促进作用;Western blot法检测其对细胞内精氨酸酶1(Arg1)、P53、基质金属蛋白酶2(MMP-2)以及上皮钙黏素(ECD)蛋白表达的影响;实时定量PCR检测诱导型一氧化氮合酶(i NOS)m RNA的表达;Griess法检测各组HepG2细胞产生的一氧化氮(NO)浓度;TranswellTM实验及细胞划痕实验测定HepG2细胞的侵袭、迁移能力。结果 nor-NOHA抑制肝癌细胞的增殖并诱导其凋亡;nor-NOHA降低Arg1、MMP-2的表达水平,升高P53、ECD的水平并增加NO的合成;nor-NOHA降低肝癌细胞的侵袭、迁移能力。结论nor-NOHA通过抑制Arg1,可诱导肝癌细胞凋亡并降低其侵袭、迁移能力,其机制与升高i NOS的表达,产生高浓度NO有关。
Objective To investigate the inhibitory effect of arginase inhibitor nor-NOHA on HepG2 hepatocarcinoma cells and its mechanism. Methods CCK-8 and flow cytometry were used to detect the effects of nor-NOHA on the proliferation and apoptosis of human hepatocellular carcinoma HepG2 cells. The results of Western blot (Arg1), P53, MMP-2, and E-cadherin (ECD) in cells were detected by real-time PCR. The expressions of inducible nitric oxide synthase NOS) m RNA expression. The concentration of nitric oxide (NO) produced by HepG2 cells was detected by Griess method. The invasion and migration of HepG2 cells were determined by TranswellTM assay and cell scratch assay. Results NO-NOR could inhibit the proliferation and induce the apoptosis of hepatocellular carcinoma cells; nor-NOHA decreased the expression of Arg1 and MMP-2, increased the levels of P53 and ECD and increased the NO synthesis; nor-NOHA reduced the invasion of hepatocellular carcinoma cells, Migration ability. Conclusion Nor-NOHA can induce hepatocellular carcinoma cell apoptosis and reduce its invasion and migration by inhibiting Arg1, and its mechanism is related to increasing i NOS expression and producing high concentration of NO.