El-Ghonemy Dina H, Ali Sanaa A, Abdel-Megeed Rehab M, Elshafei Ali M
Microbial Chemistry Department, Biotechnology Research Institute, National Research Centre, 33 El Buhouth St, Giza, EG-12622, Egypt.
Therapeutic Chemistry Department, Pharmaceutical and Drug Industries Research Institute, National Research Center, 33 El Buhouth St., Giza, EG-12622, Egypt.
J Genet Eng Biotechnol. 2023 Mar 30;21(1):38. doi: 10.1186/s43141-023-00493-x.
Hepatocellular carcinoma (HCC) is among the common cancers, but difficult to diagnose and treat. L-asparaginase has been introduced in the treatment protocol of pediatric acute lymphoblastic leukemia (ALL) since the 1960s with a good outcome and increased survival rates to nearly 90%. Moreover, it has been found to have therapeutic potential in solid tumors. Production of glutaminase-free-L-asparaginase is of interest to avoid glutaminase-related toxicity and hypersensitivity. In the current study, an extracellular L-asparaginase that is free of L-glutaminase was purified from the culture filtrate of an endophytic fungus Trichoderma viride. The cytotoxic effect of the purified enzyme was evaluated in vitro against a panel of human tumor cell lines and in vivo against male Wister albino mice intraperitoneally injected with diethyl nitrosamine (200 mg/kg bw), followed by (after 2 weeks) oral administration of carbon tetrachloride (2 mL/kg bw). This dose was repeated for 2 months, and after that, the blood samples were collected to estimate hepatic and renal injury markers, lipid profiles, and oxidative stress parameters.
L-asparaginase was purified from T. viride culture filtrate with 36 purification folds, 688.1 U/mg specific activity, and 38.9% yield. The highest antiproliferative activity of the purified enzyme was observed against the hepatocellular carcinoma (Hep-G2) cell line, with an IC of 21.2 g/mL, which was higher than that observed for MCF-7 (IC 34.2 g/mL). Comparing the DENA-intoxicated group to the negative control group, it can be demonstrated that L-asparaginase adjusted the levels of the liver function enzymes and the hepatic injury markers that had previously changed with DENA intoxication. DENA causes kidney dysfunction and altered serum albumin and creatinine levels as well. Administration of L-asparaginase was found to improve the levels of the tested biomarkers including kidney and liver function tests. L-asparaginase treatment of the DENA-intoxicated group resulted in a significant improvement in the liver and kidney tissues to near normal similar to the healthy control group.
The results suggest that this purified T. viride L-asparaginase may be able to delay the development of liver cancer and may be used as a potential candidate for future application in medicine as an anticancer medication.
肝细胞癌（HCC）是常见癌症之一，但诊断和治疗困难。自20世纪60年代以来，L-天冬酰胺酶已被引入小儿急性淋巴细胞白血病（ALL）的治疗方案，效果良好，生存率提高至近90%。此外，已发现其在实体瘤中具有治疗潜力。生产无谷氨酰胺酶的L-天冬酰胺酶有助于避免谷氨酰胺酶相关的毒性和超敏反应。在本研究中，从内生真菌绿色木霉的培养滤液中纯化出一种不含L-谷氨酰胺酶的细胞外L-天冬酰胺酶。在体外评估纯化酶对一组人类肿瘤细胞系的细胞毒性作用，在体内评估其对腹腔注射二乙基亚硝胺（200 mg/kg体重）的雄性Wistar白化小鼠的作用，2周后口服给予四氯化碳（2 mL/kg体重）。该剂量重复2个月，之后采集血样以评估肝和肾损伤标志物、血脂谱和氧化应激参数。
从绿色木霉培养滤液中纯化出L-天冬酰胺酶，纯化倍数为36，比活性为688.1 U/mg，产率为38.9%。纯化酶对肝癌（Hep-G2）细胞系的抗增殖活性最高，IC50为21.2 μg/mL，高于MCF-7细胞系（IC50为34.2 μg/mL）。将二乙基亚硝胺中毒组与阴性对照组比较，可以证明L-天冬酰胺酶调节了先前因二乙基亚硝胺中毒而改变的肝功能酶和肝损伤标志物水平。二乙基亚硝胺还会导致肾功能障碍，并改变血清白蛋白和肌酐水平。发现给予L-天冬酰胺酶可改善包括肝肾功能检测在内的受试生物标志物水平。L-天冬酰胺酶治疗二乙基亚硝胺中毒组导致肝和肾组织显著改善，接近正常，类似于健康对照组。
结果表明，这种纯化的绿色木霉L-天冬酰胺酶可能能够延缓肝癌的发展，并可能作为未来医学应用中的一种潜在抗癌药物候选物。
