| 晁延军,李英,张大闯,等.白细胞介素 -18抑制肝癌细胞增殖、促进细胞凋亡的机制研究[J].安徽医药,2021,25(3):446-450. |
| 白细胞介素 -18抑制肝癌细胞增殖、促进细胞凋亡的机制研究 |
| Mechanism of IL-18 inhibiting proliferation and promoting apoptosis of hepatoma cells |
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| DOI：10.3969/j.issn.1009-6469.2021.03.005 |
| 中文关键词: 肝肿瘤 Wnt信号通路 β连环素 白细胞介素 -18 增殖 凋亡 |
| 英文关键词: Liver neoplasms Wnt signaling pathway Beta catenin Interleukin-18 Proliferation Apoptosis |
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| 中文摘要: |
| 目的探讨白细胞介素 -18(IL-18)通过调控 Wnt/β-连环素( β-catenin)信号通路影响肝癌细胞增殖及凋亡的作用机制。方法培养肝癌细胞株 HepG2、Bel-7402与正常肝细胞株 L02，实时荧光定量逆转录聚合酶链反应( qRT-PCR)与蛋白质印迹法(Western blotting)分别检测细胞 IL-18 mRNA及蛋白表达。预实验中筛选出 HepG2细胞为实验细胞，分别转染 pcDNA、pcDNAIL-18，Western blotting验证转染效率。四甲基偶氮唑盐微量酶反应比色法( MTT法)与膜联蛋白 V(Annexin V)-异硫氰酸荧光素( FITC)/碘化丙啶( PI)染色实验分别检测 HepG2细胞增殖及凋亡情况。 qRT-PCR与 Western blotting分别检测 HepG2细胞中 B淋巴细胞瘤 -2相关 X蛋白( Bax)、 B淋巴细胞瘤 -2(Bcl-2)、β-catenin、细胞周期蛋白 D1(Cyclin D1)表达水平。 HepG2细胞中转染 IL-18过表达质粒后再加入 Wnt/β-catenin通路激活剂(SKL2001)，并检测其对 HepG2细胞增殖及凋亡的影响。结果与 L02细胞相比，肝癌 HepG2、Bel-7402细胞中 IL-18 mRNA及蛋白表达均显著降低［( 1.02±0.12)比( 0.39±0.09)(/ 0.45±0.12)；(0.41±0.14)比( 0.11±0.13)(/ 0.19±0.14)］(P<0.05)； IL-18过表达后 HepG2细胞吸光度［24 h(0.31±0.10)比( 0.27±0.13)； 48 h(0.63±0.12)比(0.39±0.14)； 72 h(0.98±0.18)比( 0.54±0.15)］Bcl-2及 β-catenin、Cyclin D1蛋白表达水平均明显低于 pcDNA组，而细胞凋亡率［( 6.93±0.22)%比( 20.54±3.97)%］及 Bax蛋白达水平均明显升高； SKL2001可激活 Wnt/β-catenin信号通路进而逆转 IL-18对肝癌细胞增殖的抑制作用及其对细胞凋亡的促进作用。结论 IL-18可下调肝癌细胞 β-catenin表达而抑制 Wnt/β |
| 英文摘要: |
| Objective To investigate the mechanism of interleukin-18 (IL-18) affecting the proliferation and apoptosis of hepatoma cells by regulating Wnt/β-catenin signaling pathway.Methods Hepatoma cell lines HepG2, Bel-7402 and normal liver cell line L02 were cultured, and IL-18 mRNA and protein expression were detected by qRT-PCR and Western blotting, respectively. In the prelimi?nary experiment, HepG2 cells were selected as experimental cells, transfected with pcDNA and pcDNA-IL-18, respectively, and the transfection efficiency was verified by Western blotting. MTT and Annexin V-FITC/PI staining experiments were used to detect the pro? liferation and apoptosis of HepG2 cells. The expression levels of Bax, Bcl-2, β-catenin and Cyclin D1 in HepG2 cells were detected by qRT-PCR and Western blotting, respectively. HepG2 cells were transfected with IL-18 overexpression plasmid and then Wnt/β-catenin pathway activator (SKL2001) was added to detect the proliferation and apoptosis of HepG2 cells.Results Compared with L02 cells, IL18 mRNA and protein expression in HepG2 and Bel-7402 cells were significantly decreased [(1.02±0.12) vs. (0.39±0.09)/(0.45±0.12); (0.41±0.14) vs. (0.11±0.13)/(0.19±0.14)] (P<0.05). After IL-18 overexpression, the OD value of HepG2 cells [24 h (0.31±0.10) vs. (0.27±0.13);48 h (0.63±0.12) vs. (0.39±0.14);72 h (0.98±0.18) vs. (0.54±0.15)] and the expression levels of Bcl-2, β-catenin and Cyclin D1 were significantly lower than those of pcDNA group, while the apoptosis rate [(6.93±0.22)% vs. (20.54±3.97)%] and Bax protein ex? pression level were significantly increased. SKL2001 could activate Wnt/β-catenin signaling pathway to reverse the inhibitory effect of IL-18 on hepatoma cell proliferation and its promotion of apoptosis.Conclusion IL-18 can down-regulate the expression of β-catenin in hepatocellular carcinoma cells and inhibit Wnt/β-catenin signaling pathway, thereby attenuating the proliferation of hepatoma cells and increasing the level of apoptosis. |
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