基于ULK1/Atg13信号通路探究白皮杉醇对肝癌HepG2细胞增殖、凋亡、自噬的影响

摘要/Abstract

摘要： 目的探究白皮杉醇对肝癌HepG2细胞增殖、凋亡、自噬的调控作用及UNC-51样激酶1(ULK1)/自噬相关蛋白13(Atg13)信号通路的影响。方法将体外培养至对数生长期的肝癌HepG2细胞分为对照组(常规培养HepG2细胞)、顺铂组(含HepG2细胞的培养基中加入顺铂10 μmol/L)、白皮杉醇低、中、高浓度组(含HepG2细胞的培养基中加入白皮杉醇10、20、40 μmol/L),培养72 h后。四甲基偶氮唑蓝法、流式细胞术、单丹磺酰戊二胺染色分别检测HepG2细胞增殖率、凋亡率、自噬情况;荧光定量PCR和蛋白印迹法分别检测HepG2细胞中微管相关蛋白轻链3Ⅱ(LC3Ⅱ)、ULK1、Atg13、B淋巴细胞瘤-2相关X蛋白(Bax)信使RNA(mRNA)和蛋白水平。结果对照组HepG2细胞未形成自噬小体;顺铂组HepG2细胞自噬小体大量形成;白皮杉醇低、中、高浓度组HepG2细胞自噬小体形成数量依次增多,但不及顺铂组。与对照组相比,顺铂组、白皮杉醇低、中、高浓度组HepG2细胞增殖率显著降低(P<0.05),凋亡率、LC3Ⅱ、ULK1、Atg13、Bax mRNA和蛋白水平显著升高(P<0.05);与顺铂组相比,白皮杉醇低、中、高浓度组HepG2细胞增殖率显著升高(P<0.05),凋亡率、LC3Ⅱ、ULK1、Atg13、Bax mRNA和蛋白水平显著降低(P<0.05);与白皮杉醇低浓度组相比,白皮杉醇中、高浓度组HepG2细胞增殖率依次降低(P<0.05),凋亡率、LC3Ⅱ、ULK1、Atg13、Bax mRNA和蛋白水平依次升高(P<0.05)。结论白皮杉醇能抑制HepG2细胞增殖,促进HepG2细胞凋亡和自噬,其机制可能与激活ULK1/Atg13信号通路有关。

关键词: 白皮杉醇, 肝癌HepG2细胞, UNC-51样激酶1, 自噬相关蛋白13, 增殖, 凋亡, 自噬

Abstract: Objective To investigate the regulatory effects of piceatannol on proliferation, apoptosis and autophagy of liver cancer HepG2 cells, as well as its effects on the UNC-51-like kinase 1 (ULK1)/autophagy associated protein 13 (Atg13) signaling pathway. Methods HepG2 cells cultured in vitro until the logarithmic growth phase were divided into control group (HepG2 cells were conventionally cultured), cisplatin group (add 10 μmol/L of cisplatin to the culture medium containing HepG2 cells) and low piceatannol (add 10 μmol/L of piceatannol to the culture medium containing HepG2 cells), medium piceatannol (add 20 μmol/L of piceatannol to the culture medium containing HepG2 cells), high piceatannol (add 40 μmol/L of piceatannol to the culture medium containing HepG2 cells) groups. The cells in all groups were cultured for 72 hours. The proliferation rate, apoptosis rate and autophagy of HepG2 cells were detected by methyl thiazolyl tetrazolium assay, flow cytometry and monodansyl pentanediamine staining, respectively. The messenger RNA (mRNA) and protein levels of microtubule-associated protein light chain 3Ⅱ (LC3Ⅱ), ULK1, Atg13, B-lymphoblastoma-2-associated X protein (Bax) in HepG2 cells were detected by fluorescence quantitative PCR and western blot, respectively. Methods The control group HepG2 cells did not form autophagosomes. A large number of autophagosomes were formed in HepG2 cells of the cisplatin group. The number of autophagosome formation in HepG2 cells in the piceatannol low, medium, and high concentration groups were increased sequentially, but was not as high as that in the cisplatin group. Compared with control group, the proliferation rate of HepG2 cells in cisplatin group, piceatannol low, medium and high concentration groups was significantly decreased (P<0.05), and the apoptosis rate, the mRNA and protein levels of LC3Ⅱ, ULK1, Atg13 and Bax were significantly increased (P<0.05). Compared with cisplatin group, the proliferation rate of HepG2 cells in piceatannol low, medium and high concentration groups was significantly increased (P<0.05), and the apoptosis rate, the mRNA and protein levels of LC3Ⅱ, ULK1, Atg13 and Bax were significantly decreased (P<0.05). Compared with piceatannol low concentration group, the proliferation rate of HepG2 cells in piceatannol medium and high concentration groups was decreased in turn (P<0.05), and the apoptosis rate, the mRNA and protein levels of LC3Ⅱ, ULK1, Atg13 and Bax were increased in turn (P<0.05). Conclusion Piceatannol can inhibit proliferation of HepG2 cells, promote apoptosis and autophagy of HepG2 cells, and its mechanism may be related to the activation of ULK1/Atg13 signaling pathway.

Key words: Piceatannol, Liver cancer HepG2 cells, UNC-51-like kinase 1, Autophagy associated protein 13, Proliferation, Apoptosis, Autophagy

张沙沙, 刘改玲, 周红霞. 基于ULK1/Atg13信号通路探究白皮杉醇对肝癌HepG2细胞增殖、凋亡、自噬的影响[J]. 肝脏, 2025, 30(1): 61-64.

ZHANG Sha-sha, LIU Gai-ling, ZHOU Hong-xia. The effects of piceatannol on proliferation, apoptosis and autophagy of liver cancer HepG2 cells were investigated based on ULK1/Atg13 signaling pathway[J]. Chinese Hepatolgy, 2025, 30(1): 61-64.

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