胰蛋白酶 K 蛋白酶（TKP）是一种来自的丝氨酸蛋白酶，通过阻断肝癌细胞的有氧糖酵解来抑制细胞增殖。

TKP, a Serine Protease from , Inhibits Cell Proliferation by Blocking Aerobic Glycolysis in Hepatocellular Carcinoma Cells.

机构信息

Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of National Ministry of Education, Shanxi University, Taiyuan, China.

出版信息

Nutr Cancer. 2022;74(1):333-345. doi: 10.1080/01635581.2021.1882508. Epub 2021 Feb 5.

DOI:10.1080/01635581.2021.1882508

PMID:33544002

Abstract

AIM

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. TKP is a serine protease extracted from the fruit of . We investigated the impact of TKP on the proliferation of HCC cells and its underlying mechanisms.

METHODS

Bel-7402 and HepG2 cell viability and colony formation capacity were evaluated using MTT and colony formation assays, respectively. Glucose uptake and lactate production were determined using glucose and lactate assay kits. The mRNA expressions of GLUT1, PDK, LDHA, PKM2, β-catenin, c-Myc, and HnRNPA1 were assessed using real-time PCR analysis. Protein expression and the distribution of PKM2 were examined by western blot assay.

RESULTS

TKP significantly inhibited Bel-7402 and HepG2 cell survival and colony formation capacity. The IC50 values of TKP against Bel-7402 and HepG2 cells were 31.37 ± 1.33 and 27.41 ± 0.81 μg/mL, respectively. TKP restrained aerobic glycolysis. TKP decreased the expression level, nuclear protein level and pyruvate kinase activity of PKM2, whereas overexpression PKM2 reversed the suppression of TKP on glycolysis. TKP inhibited the β-catenin/c-Myc/HnRNPA1 pathway. LiCl treatment partly rescued the inhibitory effects of TKP on PKM2, aerobic glycolysis, and cell viability.

CONCLUSION

TKP suppresses HCC cell proliferation via blocking PKM2-dependent glycolysis, which is regulated by inhibiting the β-catenin/c-Myc/HnRNPA1 pathway.

摘要

目的

肝细胞癌（HCC）是最常见的恶性肿瘤之一。TKP 是一种从果实中提取的丝氨酸蛋白酶。我们研究了 TKP 对 HCC 细胞增殖的影响及其潜在机制。

方法

通过 MTT 和集落形成测定分别评估 Bel-7402 和 HepG2 细胞活力和集落形成能力。使用葡萄糖和乳酸测定试剂盒测定葡萄糖摄取和乳酸生成。通过实时 PCR 分析评估 GLUT1、PDK、LDHA、PKM2、β-catenin、c-Myc 和 HnRNPA1 的 mRNA 表达。通过 Western blot 分析检测 PKM2 的蛋白表达和分布。

结果

TKP 显著抑制 Bel-7402 和 HepG2 细胞的存活和集落形成能力。TKP 对 Bel-7402 和 HepG2 细胞的 IC50 值分别为 31.37±1.33μg/mL 和 27.41±0.81μg/mL。TKP 抑制有氧糖酵解。TKP 降低了 PKM2 的表达水平、核蛋白水平和丙酮酸激酶活性，而过表达 PKM2 逆转了 TKP 对糖酵解的抑制作用。TKP 抑制了 β-catenin/c-Myc/HnRNPA1 通路。LiCl 处理部分挽救了 TKP 对 PKM2、有氧糖酵解和细胞活力的抑制作用。