Department of Pharmacy, Henan Provincial People Hospital, Department of Pharmacy of Central China Fuwai Hospital, Central China Fuwai Hospital of Zhengzhou University, Zhengzhou, 450003 Henan China.
Cell Mol Biol Lett. 2020 Jun 9;25:36. doi: 10.1186/s11658-020-00229-6. eCollection 2020.
Rhomboid domain containing 1 (RHBDD1) plays a crucial role in tumorigenesis. Silibinin, which is a natural extract from milk thistle, has shown anti-tumor effects against various tumors. Here, we investigate whether silibinin affects the function of RHBDD1 in non-small cell lung cancer (NSCLC) cell proliferation, migration and invasion.
The Oncomine database and an immunohistochemistry (IHC) assay were used to determine the RHBDD1 expression levels in lung cancer tissues. The associations between RHBDD1 and overall survival rate or clinicopathological parameters were respectively assessed using the Kaplan-Meier overall survival analysis or Chi-squared test. CCK-8 and Transwell assays were applied to analyze cell proliferation, migration and invasion. A549 cells were incubated with increasing concentrations of silibinin. RHBDD1 knockdown and overexpression were achieved via transfection with si-RHBDD1 or RHBDD1 overexpression plasmid, respectively. Western blotting was performed to measure the expressions of epithelial-mesenchymal transition (EMT) markers.
We found that overexpression of RHBDD1 in lung cancer tissues correlates with a poor prognosis of survival. Clinical specimen analysis showed that upregulation of RHBDD1 correlates remarkably well with TNM stage and lymph node metastasis. Silibinin suppresses A549 cell proliferation, migration, invasion and EMT in a dose-dependent manner. Importantly, RHBDD1 was downregulated in silibinin-treated A549 cells. RHBDD1 overexpression reversed the suppressive effects of silibinin on A549 cell proliferation, migration, invasion and EMT expression, while its knockdown enhanced them.
These findings shown an anti-tumor impact of silibinin on NSCLC cells via repression of RHBDD1.
菱形结构域包含 1 型（RHBDD1）在肿瘤发生中起着至关重要的作用。水飞蓟宾是奶蓟草的一种天然提取物，已显示出对各种肿瘤的抗肿瘤作用。在这里，我们研究了水飞蓟宾是否会影响非小细胞肺癌（NSCLC）细胞增殖、迁移和侵袭中 RHBDD1 的功能。
使用 Oncomine 数据库和免疫组织化学（IHC）检测来确定肺癌组织中 RHBDD1 的表达水平。使用 Kaplan-Meier 总体生存分析或卡方检验分别评估 RHBDD1 与总生存率或临床病理参数之间的关联。CCK-8 和 Transwell 测定用于分析细胞增殖、迁移和侵袭。用递增浓度的水飞蓟宾孵育 A549 细胞。通过转染 si-RHBDD1 或 RHBDD1 过表达质粒来实现 RHBDD1 的敲低和过表达。通过 Western blot 测定来测量上皮-间充质转化（EMT）标志物的表达。
我们发现，肺癌组织中 RHBDD1 的过表达与生存率较差相关。临床标本分析表明，RHBDD1 的上调与 TNM 分期和淋巴结转移显著相关。水飞蓟宾以剂量依赖性方式抑制 A549 细胞的增殖、迁移、侵袭和 EMT。重要的是，水飞蓟宾处理的 A549 细胞中 RHBDD1 的表达下调。RHBDD1 的过表达逆转了水飞蓟宾对 A549 细胞增殖、迁移、侵袭和 EMT 表达的抑制作用，而其敲低则增强了这些作用。
这些发现表明，水飞蓟宾通过抑制 RHBDD1 对 NSCLC 细胞发挥抗肿瘤作用。
