Department of Biochemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt.
Department of Biochemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt.
Life Sci. 2020 Aug 1;254:117794. doi: 10.1016/j.lfs.2020.117794. Epub 2020 May 15.
MicroRNAs (miRNAs) are non-coding RNAs that control post-transcriptional gene expression. Recently, miRNAs were confirmed to be promising biomarkers for different pathological conditions. This study assessed the role of serum miR-16 and miR-375 in HCC development in chronic liver disease patients such as cirrhosis. Moreover, miR-16 and miR-375 levels were estimated in HCC cell lines (HepG2 and Huh7) after treatment with doxorubicin (DOX), thymoquinone (TQ) and their combination.
Serum miR-16 and miR-375 were analyzed in 30 HCC patients, 20 cirrhosis patients and 10 healthy volunteers using RT-PCR. Moreover, HepG2 and Huh7 cells were incubated with DOX, TQ or TQ/DOX combination for 24 h and the levels of miR-16, miR-375 and gene expression of anti-apoptotic protein BCL-2 were determined in cell lysates using RT- PCR. Moreover, the ability of DOX, TQ and TQ/DOX combination to induce apoptosis were analyzed by measuring caspase-3 expression using ELISA method.
Serum miR-16 and miR-375 levels were significantly decreased in HCC patients as compared to cirrhosis and healthy control group. Also, combined use of serum miR-16 and miR-375 showed a better predictive ability than each alone. Moreover, the expression level of miR-16 and miR-375 in HepG2 and Huh7 cells increased significantly after treatment with DOX and TQ. Also, TQ/DOX combination improved apoptosis by increasing caspase-3 expression and decreasing of BCL-2 expression.
This study proved that the combined use of serum miR-16 and miR-375 was better than each alone for HCC detection. Moreover, TQ induced apoptosis and upregulatedmiR-16 and miR-375 expression in HCC cells that may explain its anticancer activity.
微小 RNA（miRNA）是一种非编码 RNA，可控制转录后基因表达。最近，miRNA 被证实是不同病理状态下有前途的生物标志物。本研究评估了血清 miR-16 和 miR-375 在慢性肝病患者（如肝硬化）肝癌发展中的作用。此外，在用多柔比星（DOX）、百里醌（TQ）及其组合处理 HepG2 和 Huh7 肝癌细胞系后，估计了 miR-16 和 miR-375 的水平。
使用 RT-PCR 分析 30 例 HCC 患者、20 例肝硬化患者和 10 例健康志愿者的血清 miR-16 和 miR-375。此外，用 DOX、TQ 或 TQ/DOX 组合孵育 HepG2 和 Huh7 细胞 24 小时，并用 RT-PCR 测定细胞裂解物中 miR-16、miR-375 和抗凋亡蛋白 BCL-2 的基因表达。此外，通过酶联免疫吸附测定法测量 caspase-3 表达来分析 DOX、TQ 和 TQ/DOX 组合诱导细胞凋亡的能力。
与肝硬化和健康对照组相比，HCC 患者血清 miR-16 和 miR-375 水平显著降低。此外，联合使用血清 miR-16 和 miR-375 比单独使用每种 miRNA 具有更好的预测能力。此外，DOX 和 TQ 处理后 HepG2 和 Huh7 细胞中 miR-16 和 miR-375 的表达水平显著增加。此外，TQ/DOX 联合用药通过增加 caspase-3 表达和降低 BCL-2 表达来改善细胞凋亡。
本研究证明，联合使用血清 miR-16 和 miR-375 比单独使用每种 miRNA 更有利于 HCC 的检测。此外，TQ 诱导 HCC 细胞凋亡并上调 miR-16 和 miR-375 的表达，这可能解释了其抗癌活性。
