五味子乙素通过靶向RhoA/ROCK1通路抑制肝细胞癌的肿瘤进展。
Schisandrin B inhibits tumor progression of hepatocellular carcinoma by targeting the RhoA/ROCK1 pathway.
作者信息
Yang Hui, Wu Tengfei
机构信息
OBiO Technology (Shanghai) Corp., Ltd., Shanghai, China.
Department of Laboratory Animal Science, China Medical University, Shenyang, China.
出版信息
J Gastrointest Oncol. 2023 Apr 29;14(2):533-543. doi: 10.21037/jgo-23-87.
BACKGROUND
Schisandrin B (Sch. B) performs various pharmacological properties, including anticancer activities. However, the pharmacological mechanisms of Sch. B in hepatocellular carcinoma (HCC) are not fully elucidated. We investigated the impact and mechanism on progression in HCC, and to provide new experimental evidence for HCC treatment.
METHODS
To determine the inhibitory effect of Sch. B on HCC , 32 Balb/c nude mice were used to prepare the tumor-bearing mice model by subcutaneously inoculating HCC cells (Huh-7). As tumor volume grew to 100 mm, mice were randomly divided into Saline (control group), 100 mg/kg Sch. B group (Sch. B-L), 200 mg/kg Sch. B group (Sch. B-M), and 400 mg/kg Sch. B group (Sch. B-H) (n=8). Saline or different concentration Sch. B was used to treat mice via gavage administration for 21 days. After mice were euthanized, tumor weight and volume were evaluated. Cell apoptosis was detected by TUNEL. Ki-67 and PCNA were detected by immunohistochemical staining. The RhoA and Rho-associated protein kinase 1 (ROCK1) were determined by western blot. experiment, Huh-7 cell were treated by Sch. B at 40, 30, 20, 10, 5, 1, and 0 µM to detect cell proliferation by Cell Counting Kit-8 (CCK-8). Huh-7 cells were divided as a control group, Sch. B group, and Sch. B + RhoA overexpression (Sch. B + RhoA) group. RhoA and ROCK1 were examined. The colony formation assay and flow cytometry were used to detect cell proliferation and apoptosis. The wound healing and Transwell assays were used for cell metastasis detection.
RESULTS
Our results showed 100, 200 and 400 mg/kg Sch. B significantly reduced tumor weight and volume. And 200 and 400 mg/kg Sch. B increased apoptosis, and reduced Ki-67 and PCNA levels, inhibited the RhoA and ROCK1 (P<0.05). experiment, Sch. B inhibited Huh-7 cell proliferation at concentration more than 10 μM (P<0.05). Sch. B decreased cell duplication, promoted apoptosis and blocked migration and invasion of Huh-7 (P<0.05). Sch. B inhibited RhoA and ROCK1 level as compared with control group (P<0.05). RhoA overexpression reversed the effect of Sch. B (P<0.05).
CONCLUSIONS
Sch. B inhibits Huh-7 cells progression via RhoA/ROCK1 pathway. The results provide new evidence for the clinical treatment of HCC.
背景
五味子乙素（Sch. B）具有多种药理特性，包括抗癌活性。然而，Sch. B在肝细胞癌（HCC）中的药理机制尚未完全阐明。我们研究了其对HCC进展的影响及机制，为HCC治疗提供新的实验证据。
方法
为确定Sch. B对HCC的抑制作用，32只Balb/c裸鼠通过皮下接种HCC细胞（Huh-7）制备荷瘤小鼠模型。当肿瘤体积长至100 mm时，将小鼠随机分为生理盐水组（对照组）、100 mg/kg Sch. B组（Sch. B-L）、200 mg/kg Sch. B组（Sch. B-M）和400 mg/kg Sch. B组（Sch. B-H）（n = 8）。通过灌胃给予生理盐水或不同浓度的Sch. B治疗小鼠21天。小鼠安乐死后，评估肿瘤重量和体积。通过TUNEL检测细胞凋亡。通过免疫组织化学染色检测Ki-67和PCNA。通过蛋白质印迹法测定RhoA和Rho相关蛋白激酶1（ROCK1）。在实验中，用40、30、20、10、5、1和0 μM的Sch. B处理Huh-7细胞，通过细胞计数试剂盒-8（CCK-8）检测细胞增殖。将Huh-7细胞分为对照组、Sch. B组和Sch. B + RhoA过表达组（Sch. B + RhoA）。检测RhoA和ROCK1。采用集落形成试验和流式细胞术检测细胞增殖和凋亡。采用伤口愈合试验和Transwell试验检测细胞转移。
结果
我们的结果显示，100、200和400 mg/kg的Sch. B显著降低了肿瘤重量和体积。200和400 mg/kg的Sch. B增加了细胞凋亡，降低了Ki-67和PCNA水平，抑制了RhoA和ROCK1（P < 0.05）。在实验中，Sch. B在浓度超过10 μM时抑制Huh-7细胞增殖（P < 0.05）。Sch. B减少了细胞复制，促进了凋亡，并阻断了Huh-7的迁移和侵袭（P < 0.05）。与对照组相比，Sch. B抑制了RhoA和ROCK1水平（P < 0.05）。RhoA过表达逆转了Sch. B的作用（P < 0.05）。
结论
Sch. B通过RhoA/ROCK1途径抑制Huh-7细胞进展。这些结果为HCC的临床治疗提供了新的证据。
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