Mofed Dina, Sabet Salwa, Baiomy Ahmed A, Salem Tamer Z
Molecular Biology and Virology Lab, Biomedical Sciences Program, Zewail City of Science and Technology, October Gardens, 6th of October City, Giza 12578, Egypt.
Zoology Graduate Program, Department of Zoology, Faculty of Science, Cairo University, Giza 12613, Egypt.
Curr Issues Mol Biol. 2022 Aug 12;44(8):3632-3647. doi: 10.3390/cimb44080249.
Long non-coding RNAs (lncRNAs) are regulated in cancer cells, including lncRNA MEG3, which is downregulated in Hepatocellular Carcinoma (HCC). In addition, hepatitis C virus (HCV) core proteins are known to dysregulate important cellular pathways that are linked to HCC development. In this study, we were interested in evaluating the overexpression of lncRNA MEG3, either alone or in combination with two forms of HCV core protein (C173 and C191) in HepG2 cells. Cell viability was assessed by MTT assay. Transcripts' levels of key genes known to be regulated in HCC, such as , , miRNA152, , and were measured by qRT-PCR. Protein expression levels of caspase-3 and MKI67 were determined by immunocytochemistry and apoptosis assays. The co-expression of lncRNA MEG3 and C191 resulted in a marked increase and accumulation of dead cells and a reduction in cell viability. In addition, a marked increase in the expression of tumor suppressor genes ( and miRNA152), as well as a marked decrease in the expression of oncogenes (, BCL2, and ), were detected. Moreover, apoptosis assay results revealed a significant increase in total apoptosis (early and late). Finally, immunocytochemistry results detected a significant increase in apoptotic marker caspase-3 and a decrease in tumor marker MKI67. In this study, transgene expression of C191 and lncRNA MEG3 showed induction in apoptosis in HepG2 cells greater than the expression of each one alone. These results suggest potential anticancer characteristics.
长链非编码RNA（lncRNAs）在癌细胞中受到调控，包括lncRNA MEG3，其在肝细胞癌（HCC）中表达下调。此外，已知丙型肝炎病毒（HCV）核心蛋白会失调与HCC发展相关的重要细胞通路。在本研究中，我们感兴趣的是评估lncRNA MEG3单独或与两种形式的HCV核心蛋白（C173和C191）在HepG2细胞中的过表达情况。通过MTT法评估细胞活力。通过qRT-PCR检测已知在HCC中受到调控的关键基因的转录水平，如 、 、miRNA152、 和 。通过免疫细胞化学和凋亡检测确定caspase-3和MKI67的蛋白表达水平。lncRNA MEG3和C191的共表达导致死细胞显著增加和积累，以及细胞活力降低。此外，检测到肿瘤抑制基因（ 和miRNA152）的表达显著增加，以及癌基因（ 、BCL2和 ）的表达显著降低。此外，凋亡检测结果显示总凋亡（早期和晚期）显著增加。最后，免疫细胞化学结果检测到凋亡标志物caspase-3显著增加，肿瘤标志物MKI67降低。在本研究中，C191和lncRNA MEG3的转基因表达在HepG2细胞中显示出比单独表达其中任何一种更强的诱导凋亡作用。这些结果表明其具有潜在的抗癌特性。
