Department of Anorectal Surgery, Jining No.1 People's Hospital, Jining 272011, China; Affiliated Jining No.1 People's Hospital of Jining Medical University, Jining Medical University, Jining 272067, China.
Department of Anorectal Surgery, Jining No.1 People's Hospital, Jining 272011, China.
Exp Mol Pathol. 2019 Feb;106:131-138. doi: 10.1016/j.yexmp.2019.01.003. Epub 2019 Jan 8.
Colorectal cancer (CRC) is a troublesome disease with high morbidity and mortality. Ginsenoside Rg3 possesses anti-cancer properties. Colon Cancer Associated Transcript 1 (CCAT1) participates in the genesis, development, invasion and metastasis of colorectal cancer. In our study, we explored the effects of Rg3 on CRC cell line Caco-2 by regulating CCAT1.
CRC tissue was obtained from hospital and Caco-2 cells were purchased. Caco-2 cells were treated with Rg3 and/or transfected with pc- CCAT1 or pcDNA3.1. The group without Rg3 treatment was treated as control. Cell viability, cell apoptosis, cell migration and invasion were detected by Cell Counting Kit-8 assay, flow cytometry and Transwell chamber migration/invasion assay, respectively. The expression of CyclinD1, apoptosis related proteins (p53, Bcl-2, Bax, pro-/Cleaved-Caspase-3), migration and invasion related proteins (MMP-9 and vimentin), and phosphatidylinositol 3'-kinase (PI3K)/protein kinase B (AKT) related proteins (p/t-PI3K, p/t-AKT) were examined by western blot. The expression of CCAT1 was measured by quantitative real time RCR (qRT-PCR).
Rg3 significantly decreased cell viability, migration and invasion, and promoted apoptosis. Meanwhile, the expression of Cyclin D1, matrix metalloproteinase (MMP)-9 and vimentin was downregulated. The expression of apoptosis-related proteins p53, Bax, and Cleaved-Caspase-3 were upregulated while Bcl-2 was downregulated by the treatment of Rg3 compared with control. Furthermore, CCAT1 was upregulated in CRC tissue and Rg3 negatively regulated CCAT1 expression. Transfection with pc-CCAT1 led to the opposite results as compared with transfection with pcDNA3.1 in Rg3 treated cells. In addition, Rg3 decreased the phosphorylation of PI3K and AKT.
Ginsenoside Rg3 inhibits migration and invasion, and promotes apoptosis of Caco-2 cells by suppression expression of LncRNA CCAT1.
结直肠癌（CRC）是一种发病率和死亡率都很高的棘手疾病。人参皂苷 Rg3 具有抗癌特性。结肠癌相关转录物 1（CCAT1）参与结直肠癌的发生、发展、侵袭和转移。在我们的研究中，我们通过调节 CCAT1 来探索 Rg3 对 CRC 细胞系 Caco-2 的影响。
从医院获取 CRC 组织并购买 Caco-2 细胞。用 Rg3 处理 Caco-2 细胞和/或用 pc-CCAT1 或 pcDNA3.1 转染。未用 Rg3 处理的组作为对照。用细胞计数试剂盒-8 测定法、流式细胞术和 Transwell 室迁移/侵袭测定法分别检测细胞活力、细胞凋亡、细胞迁移和侵袭。用 Western blot 检测细胞周期蛋白 D1、凋亡相关蛋白（p53、Bcl-2、Bax、pro-/Cleaved-Caspase-3）、迁移和侵袭相关蛋白（MMP-9 和波形蛋白）以及磷脂酰肌醇 3'-激酶（PI3K）/蛋白激酶 B（AKT）相关蛋白（p/t-PI3K、p/t-AKT）的表达。用实时定量 PCR（qRT-PCR）测量 CCAT1 的表达。
Rg3 显著降低细胞活力、迁移和侵袭，并促进凋亡。同时，下调细胞周期蛋白 D1、基质金属蛋白酶（MMP）-9 和波形蛋白的表达。与对照组相比，Rg3 处理后凋亡相关蛋白 p53、Bax 和 Cleaved-Caspase-3 的表达上调，而 Bcl-2 的表达下调。此外，CRC 组织中上调 CCAT1，Rg3 负调节 CCAT1 的表达。与 pcDNA3.1 转染相比，pc-CCAT1 转染导致 Rg3 处理细胞的结果相反。此外，Rg3 降低了 PI3K 和 AKT 的磷酸化。
人参皂苷 Rg3 通过抑制长链非编码 RNA CCAT1 的表达抑制 Caco-2 细胞的迁移和侵袭，并促进细胞凋亡。
