Jiang Zhi Tao, Han Yi, Liu Xiao Yan, Lv Ling Yan, Pan Jin Huo, Liu Chun Di
Department of Pharmacy Office, Zhangjiagang Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Zhangjiagang, Jiangsu, People's Republic of China.
College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, People's Republic of China.
Onco Targets Ther. 2020 Apr 8;13:2973-2985. doi: 10.2147/OTT.S238074. eCollection 2020.
Triterpene has attracted considerable interests because it exhibits anticancer effects. However, the effects of tripterine on hepatocellular carcinoma (HCC) are not well studied. In the current study, the mechanism of tripterine on HCC cells growth and metastasis was examined.
The inhibitory effect on the growth and aggressiveness in HCC cells was analyzed by Cell Counting Kit-8 (CCK-8), wound healing and Transwell assay. The levels of microRNA-532-5p (miR-532-5p) in HCC cells and tissues were measured using qRT-PCR. The expression of chemokine (C-X-C Motif) ligand 2 (CXCL2) was determined by Western blotting and immunohistochemistry (IHC). Luciferase reporter gene assay was used to validate the binding between miR-532-5p and CXCL2. The impact of tripterine on the growth and metastasis of HCC cells in vivo was analyzed using transplanted tumor model and experimental lung metastasis model, respectively.
We found that tripterine inhibited HCC cells proliferation, migration ability and invasion. Under tripterine treatment, the level of miR-532-5p was strikingly raised, and overexpression of miR‑532-5p reduced cell viability and metastatic-related traits. In addition, we identified CXCL2 as a target of miR-532-5p in HCC. Rescue experiments indicated that overexpression of CXCL2 restored the migration and invasive capacity of HCC cells inhibited by miR-532-5p or tripterine treatment. Finally, the tumor growth and metastatic ability of HCC MHCC97H cell in vivo were also significantly restrained by tripterine. The expression of CXCL2 was distinctly decreased and miR-532-5p level was increased by tripterine in vivo.
In conclusion, tripterine inhibits the growth, migration ability and invasiveness of HCC cells through intervening miR-532-5p/CXCL2.
三萜烯因其具有抗癌作用而备受关注。然而，雷公藤红素对肝细胞癌（HCC）的影响尚未得到充分研究。在本研究中，我们探讨了雷公藤红素对肝癌细胞生长和转移的作用机制。
采用细胞计数试剂盒-8（CCK-8）、伤口愈合实验和Transwell实验分析雷公藤红素对肝癌细胞生长和侵袭能力的抑制作用。运用qRT-PCR检测肝癌细胞和组织中微小RNA-532-5p（miR-532-5p）的水平。通过蛋白质免疫印迹法和免疫组织化学（IHC）检测趋化因子（C-X-C基序）配体2（CXCL2）的表达。采用荧光素酶报告基因实验验证miR-532-5p与CXCL2之间的结合。分别利用移植瘤模型和实验性肺转移模型分析雷公藤红素对肝癌细胞体内生长和转移的影响。
我们发现雷公藤红素可抑制肝癌细胞的增殖、迁移能力和侵袭能力。在雷公藤红素处理下，miR-532-5p水平显著升高，过表达miR-532-5p可降低细胞活力和转移相关特性。此外，我们确定CXCL2是肝癌中miR-532-5p的靶标。挽救实验表明，过表达CXCL2可恢复受miR-532-5p或雷公藤红素处理抑制的肝癌细胞的迁移和侵袭能力。最后，雷公藤红素在体内也显著抑制了肝癌MHCC97H细胞的肿瘤生长和转移能力。雷公藤红素在体内可使CXCL2的表达明显降低，miR-532-5p水平升高。
综上所述，雷公藤红素通过干预miR-532-5p/CXCL2抑制肝癌细胞的生长、迁移能力和侵袭能力。
