Page 271 - EJMO-9-2

Eurasian Journal of
Medicine and Oncology ANGPTL8 upregulation in CRC with liver metastasis

2.2. ANGPTL8 expression analysis 5’-CTCCTT AATGTCACGCACGAT-3’ (reverse). ANGPTL8

The protein level of ANGPTL8 expression was assessed expression was determined by quantifying the comparative
using a human enzyme-linked immunosorbent assay cycle threshold (Ct) values. The relative gene expression levels
ΔCt
kit specific for ANGPTL8 (No. E11644h, EIAab, China). were calculated using the 2 method and analyzed with JMP.6
The mRNA level of ANGPTL8 expression was measured software (SAS Institute, United States of America).
using real-time quantitative polymerase chain reaction 2.3. Statistical analyses
(RT-qPCR). Total RNA was extracted from fresh-frozen
tissue samples (100 mg) using the TRISURE™/Qiazol The data distribution was assessed using the Kolmogorov–
synthesis kit (Bioline, United Kingdom) and stored at −20°C. Smirnov normality test. Comparisons between groups
The reverse transcription process was performed using the were performed using a parametric independent sample
ReverTra Ace RT-qPCR Master Mix with gDNA Remover t-test. Associations between parameters or variables were
®
(TOYOBO, Japan). The RT-qPCR was conducted on a qTower evaluated using Pearson’s product-moment correlation test
machine (Analytic Jena, Germany) using a mix solution and univariate linear regression analysis. A significance
from the SensiFAST SYBR Green No-ROX kit (Bioline, level of 5% (p≤0.05) was applied, and the results are
United Kingdom). The forward and reverse primers for presented as mean ± SEM.
ANGPTL8 were 5’-GAGACTCAGATGGAGGAGGA-3’ and 3. Results
5’-ATGCTGCTGTGCCACCATCT-3’, respectively. β-actin
was used as the housekeeping gene, with primer sequences Based on the patients’ medical records (Figure 1), in the
of 5’- CATGTACGTTGCTATCCAGG-3’ (forward) and non-liver metastasis group, three patients had a history of

Figure 1. Characteristics of disease history in the study population across metastasis and non-metastasis groups. The graphs represent the distribution of
individuals with (Yes) and without (No) history of each condition.
Note: Asterisk (*) indicates a significant difference between “Yes” and “No” within the same group (metastasis or non-metastasis), as determined by
independent sample t-test analysis (p≤0.05).
Abbreviations: CVD: Cardiovascular disease; DM: Diabetes mellitus.

Volume 9 Issue 2 (2025) 263 doi: 10.36922/EJMO025080034