School of Laboratory Medicine, Wannan Medical College, China.
Mutat Res. 2024 Jul-Dec;829:111872. doi: 10.1016/j.mrfmmm.2024.111872. Epub 2024 Jul 6.
Among primary liver cancers, HCC is the most prevalent. Small noncoding RNAs called miRNAs control the expression of downstream target genes to take part in a variety of physiological and pathological processes, including those related to cancer.
miR-129-2-3p and SEMA4C expression levels were assessed using RT-qPCR. The CCK-8, invasion, and wound healing assays were used to confirm the capacity of HCC cells for proliferation, invasion and migration respectively. Serum SEMA4C levels were detected via ELISA. The RIP and dual-luciferase reporter assays were used to confirm the existence of intergenic binding sites. Cell apoptosis assay and cell cycle assay were performed to detect the apoptosis rate and cycle distribution of cells, and WB was performed to detect the protein expression of SEMA4C, RhoA, ROCK1, E-cadherin, N-cadherin, and vimentin. Furthermore, cancer-inhibiting role of miR-129-2-3p were further confirmed by animal tests.
miR-129-2-3p expression was reduced in HCC tissues and cells. Overexpression of miR-129-2-3p decreased the proliferation, invasion, migration, and EMT in HCC cells, whereas inhibition of miR-129-2-3p had the opposite effects. Our research also showed that SEMA4C was increased in HCC tissues, serum and cells, and that SEMA4C knockdown prevented HCC cell invasion, migration, proliferation, and EMT. Overexpression of SEMA4C reversed the inhibitory effect of miR-129-2-3p on HCC.
Overall, we discovered that through binding to SEMA4C, miR-129-2-3p regulates HCC cell proliferation, invasion, migration, and EMT.
原发性肝癌中，HCC 最为常见。miRNAs 是一类小的非编码 RNA，可以调控下游靶基因的表达，参与多种生理和病理过程，包括与癌症相关的过程。
采用 RT-qPCR 检测 miR-129-2-3p 和 SEMA4C 的表达水平。用 CCK-8 实验、侵袭实验和划痕愈合实验分别验证 HCC 细胞的增殖、侵袭和迁移能力。采用 ELISA 法检测血清 SEMA4C 水平。用 RIP 和双荧光素酶报告基因实验验证基因间结合位点的存在。通过细胞凋亡实验和细胞周期实验检测细胞的凋亡率和周期分布，通过 WB 实验检测 SEMA4C、RhoA、ROCK1、E-钙黏蛋白、N-钙黏蛋白和波形蛋白的蛋白表达。此外，还通过动物实验进一步证实了 miR-129-2-3p 的抑癌作用。
miR-129-2-3p 在 HCC 组织和细胞中的表达降低。过表达 miR-129-2-3p 可降低 HCC 细胞的增殖、侵袭、迁移和 EMT，而抑制 miR-129-2-3p 则产生相反的效果。我们的研究还表明，SEMA4C 在 HCC 组织、血清和细胞中表达增加，抑制 SEMA4C 可阻止 HCC 细胞侵袭、迁移、增殖和 EMT。过表达 SEMA4C 可逆转 miR-129-2-3p 对 HCC 的抑制作用。
总之，我们发现 miR-129-2-3p 通过与 SEMA4C 结合，调控 HCC 细胞的增殖、侵袭、迁移和 EMT。
