Tang Jian-Cai, Zhao Jia, Long Feng, Chen Jian-Ye, Mu Bo, Jiang Zhen, Ren Yonggan, Yang Jian
Department of Biochemistry.
School of Pharmacy.
J Cancer. 2018 Jan 1;9(1):32-40. doi: 10.7150/jca.21224. eCollection 2018.
Increasing evidences indicate that shikonin can suppress the tumor growth. However, the mechanisms remain elusive. In the present study, we investigated the effects and mechanisms of shikonin against esophageal cancer. The expression of hypoxia inducible factor 1α (HIF1α) and pyruvate kinase M2 (PKM2) in esophageal cancer tissues and cells was detected by immunohistochemistry and Western blot. CCK-8 was used to examine the esophageal cancer cell viability. Apoptosis and cell cycle were analyzed by flow cytometry. The expression of EGFR, PI3K, Akt, p-AKT, mTOR, HIF1α and PKM2 was detected by Western blot. EC109/pkm2 was established by lentivirus transducer. Ec109 tumor model was founded to observe the antitumor effect of shikonin . We found that HIF1α and PKM2 protein expression levels were higher in esophageal cancer tissues and cells than normal esophageal tissues and cells. Shikonin reduced esophageal cancer cells viability and induced cell cycle arrest and apoptosis. Shikonin decreased EGFR, PI3K, p-AKT, HIF1α and PKM2 expression. Overexpression of PKM2 could enhance resistance of esophageal cancer cells to shikonin. we found that shikonin reduced tumor burden, inducing cell arrest and apoptosis. Taken together, shikonin has a significant antitumor effect in the esophageal cancer by regulating HIF1α/PKM2 signal pathway.
越来越多的证据表明紫草素可以抑制肿瘤生长。然而，其机制仍不清楚。在本研究中，我们调查了紫草素抗食管癌的作用及机制。通过免疫组织化学和蛋白质印迹法检测食管癌组织和细胞中缺氧诱导因子1α（HIF1α）和丙酮酸激酶M2（PKM2）的表达。采用CCK-8检测食管癌细胞活力。通过流式细胞术分析细胞凋亡和细胞周期。通过蛋白质印迹法检测表皮生长因子受体（EGFR）、磷脂酰肌醇-3激酶（PI3K）、蛋白激酶B（Akt）、磷酸化Akt（p-AKT）、哺乳动物雷帕霉素靶蛋白（mTOR）、HIF1α和PKM2的表达。通过慢病毒转导建立EC109/pkm2细胞系。建立Ec109肿瘤模型以观察紫草素的抗肿瘤作用。我们发现食管癌组织和细胞中HIF1α和PKM2蛋白表达水平高于正常食管组织和细胞。紫草素降低食管癌细胞活力，诱导细胞周期阻滞和细胞凋亡。紫草素降低EGFR、PI3K、p-AKT、HIF1α和PKM2的表达。PKM2过表达可增强食管癌细胞对紫草素的抗性。我们发现紫草素减轻肿瘤负荷，诱导细胞停滞和凋亡。综上所述，紫草素通过调节HIF1α/PKM2信号通路对食管癌具有显著的抗肿瘤作用。
