摘要
目的探讨UNC-51样激酶1(ULK1)在肝细胞癌(HCC)中的表达及预后,敲减ULK1或抑制ULK1激酶的活性是否可以抑制肝癌细胞增殖、诱导凋亡并增强奥沙利铂的敏感性。方法在在线数据库中分析ULK1在泛癌和HCC中的表达和预后;定量实时聚合酶链反应验证不同ULK1敲减序列的敲减效率;CCK8法检测敲减ULK1对肝癌细胞系HepG2增殖能力的影响;CCK8法检测ULK1抑制剂和奥沙利铂对HepG2细胞增殖的影响;流式细胞术探讨奥沙利铂与ULK1抑制剂联合对HepG2细胞凋亡的影响;免疫印记法实验检测奥沙利铂与ULK1抑制剂联合对HepG2细胞自噬的影响。结果ULK1在HCC中高表达且与患者预后呈负相关;在HepG2细胞中成功敲减ULK1基因;敲低ULK1可降低HepG2细胞增殖能力;ULK1抑制剂和奥沙利铂联合可抑制HepG2细胞增殖,同时增加HepG2细胞凋亡数量;其机制可能与增强HepG2细胞自噬有关。结论敲低ULK1或使用ULK1抑制剂,可抑制HepG2细胞增殖并诱导凋亡,或有可能增强化疗药物毒性作用,其机制可能与细胞自噬的调控有关。
Objective To investigate the expression and prognosis of UNC-51-like kinase 1(ULK1)in hepatocellular carcinoma(HCC),and whether knocking down ULK1 or inhibiting the activity of ULK1 kinase can inhibit the proliferation of hepatocellular carcinoma cells,induce apoptosis and enhance the sensitivity of oxaliplatin.Methods The expression and prognosis of ULK1 in pan-cancer and HCC were analyzed in online database.Quantitative real-time polymerase chain reaction was used to verify the knock-down efficiency of different ULK1 knock-down sequences.CCK8 method was used to detect the effect of knockout ULK1 on the proliferation of hepatocellular carcinoma cell line HepG2.CCK8 method was used to detect the effects of ULK1 inhibitor and oxaliplatin on the proliferation of HepG2 cells.Flow cytometry was used to detect the effect of oxaliplatin combined with ULK1 inhibitor on apoptosis of HepG2 cells.Immunoblotting experiments were conducted to detect the effect of oxaliplatin combined with ULK1 inhibitors on autophagy in HepG2 cells.Results ULK1 was highly expressed in HCC and negatively correlated with the prognosis of patients.The ULK1 gene was successfully knocked down in HepG2 cells.Knocking down ULK1 can reduce the proliferation of HepG2 cells.The combination of ULK1 inhibitor and oxaliplatin can inhibit the proliferation of H epG2 cells and increase the number of apoptosis of HepG2 cells.The mechanism may be related to enhancing autophagy of HepG2 cells.Conclusion Knocking down ULK1 or using ULK1 inhibitor can inhibit the proliferation and induce apoptosis of HepG2 cells,or may enhance the toxicity of chemotherapy drugs,and its mechani sm may be related to the regulation of autophagy.
作者
毛雅珍
陈勇
林赵栋
沈玲英
林伟
MAO Yazhen;CHEN Yong;LIN Zhaodong;SHEN Lingying;LIN Wei(Department of Clinical Laboratory,Fuzhou First General Hospital,Fujian,Fuzhou 350009,China)
出处
《中国医药科学》
2024年第17期4-7,17,共5页
China Medicine And Pharmacy
基金
福州市科技计划项目(2021-S-173)。
