Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University, Yongin si 17104, Gyeonggi do, Republic of Korea.
Department of Oriental Medicinal Biotechnology, College of Life Science, Kyung Hee University, Yongin-si 17104, Gyeonggi-do, Republic of Korea.
Molecules. 2022 Nov 28;27(23):8311. doi: 10.3390/molecules27238311.
Ginsenoside Rh1 (G-Rh1), a possible bioactive substance isolated from the Korean Meyer, has a wide range of pharmacological effects. In this study, we have investigated the anticancer efficacy of G-Rh1 via in silico and in vitro methodologies. This study mainly focuses on the two metastatic regulators, Rho-associated protein kinase 1 (ROCK1) and RhoA, along with other standard apoptosis regulators. The ROCK1 protein is a member of the active serine/threonine kinase family that is crucial for many biological processes, including cell division, differentiation, and death, as well as many cellular processes and muscle contraction. The abnormal activation of ROCK1 kinase causes several disorders, whereas numerous studies have also shown that RhoA is expressed highly in various cancers, including colon, lung, ovarian, gastric, and liver malignancies. Hence, inhibiting both ROCK1 and RhoA will be promising in preventing metastasis. Therefore, the molecular level interaction of G-Rh1 with the ROCK1 and RhoA active site residues from the preliminary screening clearly shows its inhibitory potential. Molecular dynamics simulation and principal component analysis give essential insights for comprehending the conformational changes that result from G-Rh1 binding to ROCK1 and RhoA. Further, MTT assay was employed to examine the potential cytotoxicity in vitro against human lung cancer cells (A549) and Raw 264.7 Murine macrophage cells. Thus, G-Rh1 showed significant cytotoxicity against human lung adenocarcinoma (A549) at 100 µg/mL. In addition, we observed an elevated level of reactive oxygen species (ROS) generation, perhaps promoting cancer cell toxicity. Additionally, G-Rh1 suppressed the mRNA expression of RhoA, ROCK1, MMP1, and MMP9 in cancer cell. Accordingly, G-Rh1 upregulated the p53, Bax, Caspase 3, caspase 9 while Bcl2 is downregulated intrinsic pathway. The findings from our study propose that the anticancer activity of G-Rh1 may be related to the induction of apoptosis by the RhoA/ROCK1 signaling pathway. As a result, this study evaluated the functional drug-like compound G-Rh1 from in preventing and treating lung cancer adenocarcinoma via regulating metastasis and apoptosis.
人参皂苷 Rh1（G-Rh1）是一种可能从韩国 Meyer 中分离出来的生物活性物质，具有广泛的药理作用。在这项研究中，我们通过计算机模拟和体外方法研究了 G-Rh1 的抗癌功效。本研究主要集中在两个转移调节剂 Rho 相关蛋白激酶 1（ROCK1）和 RhoA 以及其他标准凋亡调节剂上。ROCK1 蛋白是活性丝氨酸/苏氨酸激酶家族的成员，对于许多生物学过程至关重要，包括细胞分裂、分化和死亡以及许多细胞过程和肌肉收缩。ROCK1 激酶的异常激活会导致多种疾病，而许多研究还表明，RhoA 在包括结肠癌、肺癌、卵巢癌、胃癌和肝癌在内的各种癌症中表达水平较高。因此，抑制 ROCK1 和 RhoA 两者将有望预防转移。因此，从初步筛选中 G-Rh1 与 ROCK1 和 RhoA 活性位点残基的分子水平相互作用清楚地表明了其抑制潜力。分子动力学模拟和主成分分析为理解 G-Rh1 与 ROCK1 和 RhoA 结合导致的构象变化提供了重要的见解。进一步，通过 MTT 测定法在体外检查了对人肺癌细胞（A549）和 Raw 264.7 鼠巨噬细胞的潜在细胞毒性。因此，G-Rh1 在 100μg/mL 时对人肺腺癌（A549）显示出显著的细胞毒性。此外，我们观察到活性氧（ROS）生成水平升高，这可能促进了癌细胞毒性。此外，G-Rh1 抑制了癌细胞中 RhoA、ROCK1、MMP1 和 MMP9 的 mRNA 表达。相应地，G-Rh1 上调了 p53、Bax、Caspase 3、Caspase 9，同时下调了 Bcl2 内在途径。本研究的结果表明，G-Rh1 的抗癌活性可能与 RhoA/ROCK1 信号通路诱导的细胞凋亡有关。因此，本研究评估了来自 的功能性药物样化合物 G-Rh1 通过调节转移和凋亡来预防和治疗肺癌腺癌的作用。
