Cancer & Radiation Countermeasures Unit, Department of Biochemistry, North-Eastern Hill University, Shillong-793022, India.
Asian Pac J Cancer Prev. 2022 Aug 1;23(8):2843-2850. doi: 10.31557/APJCP.2022.23.8.2843.
Hepatocellular carcinoma (HCC) is one of the leading drivers of cancer-related mortality in the world. As a result, researchers are constantly looking for ways to optimize the screening and diagnosis of the said malignancy.
To establish the mice model of hepatocellular carcinoma with the administration of a suitable dose of diethylnitrosamine (DEN) and examine the utility of EphA7 and pEphA7 as ideal diagnostic markers in HCC.
Swiss Albino (BALB/c) mice of around 10-12 weeks old were exposed to a known hepatocarcinogen-diethylnitrosamine at a dose of 20 mg/kg body weight at weekly intervals for a period of 4, 8, 12, & 16 weeks. Blood was collected from mice of different experimental groups, and age-matched control and serum were separated from whole blood samples. The liver homogenate was prepared after completion of treatment, and the resulting supernatant was used for enzyme assays. A range of liver biomarker enzyme assays such as Gamma-glutamyl transpeptidase (GGT), Acetylcholine esterase (AChE), GPx activity and GSH level, Heme oxygenase-1 (HO-1), GPC3 and alpha-fetoprotein (AFP) level along with the expression of Caspase-3, EphA7 and pEphA7 were evaluated.
An elevation in body weight and relative liver weight across the treatment period (4, 8, 12, 16 weeks) was observed in DEN-treated mice. Significant differences in GGT levels between control and DEN treated mice were noted in the present study (P < 0.005). In the 16th week of the treatment period, a significant difference in AchE level was noted between the treated and control group (P < 0.001). However, there was no statistically significant difference in the levels of SGOT and SGPT levels between the control and DEN treated groups (P > 0.001). Lower GSH and GPx levels were demonstrated in the treated mice as compared to control over all the treatment period. Loss of Caspase-3 expression and significant differences in expression of HO-1 activity in treated vs. non-treated group of mice were observed. Significant differences in EphA7 and pEphA7 protein expression levels were noted in the DEN-treated vs. control groups across all the treatment periods (4 weeks: P < 0.05; 8 weeks: P < 0.05; 12 weeks: P < 0.005; 16 weeks: P < 0.05).
The present study indicated that EphA7 and phosphoEphA7 over-expression might contribute to the malignancy transition, invasion development, and metastasis of HCC. As a result, along with the known markers such as AFP and others, EphA7 and pEphA7 could be a very putative biomarkers of HCC, particularly at a very early stage of cancer development.
肝细胞癌（HCC）是全球癌症相关死亡的主要驱动因素之一。因此，研究人员一直在寻找优化该恶性肿瘤筛查和诊断的方法。
用适当剂量的二乙基亚硝胺（DEN）建立肝癌小鼠模型，并研究 EphA7 和 pEphA7 作为 HCC 理想诊断标志物的效用。
将 10-12 周龄的瑞士白化（BALB/c）小鼠每周以 20mg/kg 体重的剂量暴露于已知的肝癌致癌物二乙基亚硝胺中，持续 4、8、12 和 16 周。从不同实验组的小鼠中采集血液，并从全血样本中分离血清。完成治疗后制备肝匀浆，所得上清液用于酶测定。评估了一系列肝生物标志物酶测定，如γ-谷氨酰转肽酶（GGT）、乙酰胆碱酯酶（AChE）、谷胱甘肽过氧化物酶（GPx）活性和谷胱甘肽水平、血红素加氧酶-1（HO-1）、GPC3 和甲胎蛋白（AFP）水平以及 Caspase-3、EphA7 和 pEphA7 的表达。
在 DEN 处理的小鼠中，观察到体重和相对肝重在整个治疗期间（4、8、12、16 周）升高。本研究中观察到 DEN 处理组与对照组之间 GGT 水平有显著差异（P < 0.005）。在治疗期的第 16 周，处理组和对照组之间 AchE 水平有显著差异（P < 0.001）。然而，对照组和 DEN 处理组之间的 SGOT 和 SGPT 水平无统计学差异（P > 0.001）。与对照组相比，处理组的 GSH 和 GPx 水平较低。在处理组中观察到 Caspase-3 表达的丧失和 HO-1 活性表达的显著差异。在 DEN 处理组与对照组之间，EphA7 和 pEphA7 蛋白表达水平在所有治疗期间均有显著差异（4 周：P < 0.05；8 周：P < 0.05；12 周：P < 0.005；16 周：P < 0.05）。
本研究表明 EphA7 和磷酸化 EphA7 的过度表达可能导致 HCC 的恶性转化、侵袭发展和转移。因此，除了 AFP 等已知标志物外，EphA7 和 pEphA7 可能是 HCC 的一个非常有潜力的标志物，特别是在癌症发展的早期阶段。
