Department of Radiation Oncology, The Affiliated Hospital of Qingdao University, Qingdao, Shangdong, China (mainland).
Department of Clinical Laboratory, Yuhuangding Hospital, Yantai, Shandong, China (mainland).
Med Sci Monit. 2016 May 30;22:1827-36. doi: 10.12659/msm.898801.
BACKGROUND We investigated whether the plant-derived agent triptolide (TPL) could effectively inhibit the growth and invasion of human hepatocellular carcinoma (HCC) cells. MATERIAL AND METHODS MHCC-97H cells were treated with various concentration of TPL for various times. To detect the effect of NF-κB on TPL-induced signal pathways, MHCC-97H cells were transfected with p65 siRNA or p65 cDNA, then treated with TPL. We detected cell survival and apoptosis by MTT, soft-agar colony formation assay, flow cytometry, and TUNEL assay. Cell migration and invasion was determined by Matrigel invasion and a wound-healing assay. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA); MMP-9 activity was detected by ELISA. Western blot and real-time PCR (RT-PCR) assays were used to detect p65 and MMP-9 protein and mRNA expression. A subcutaneously implanted tumor model of MHCC-97H cells in nude mice was used to assess the effects of TPL on tumorigenesis in vivo. RESULTS We showed that TPL treatment significantly suppressed growth and induced apoptosis of MHCC-97H cells in a dose- and time-dependent manner in vitro. Furthermore, TPL treatment inhibited invasion in vitro and inhibited the growth and lung metastasis of MHCC-97H cells in vivo. NF-κB and MMP-9 were inactivated with TPL treatment. Overexpression of p65 restored MMP-9 activity and inhibited the TPL anti-tumor effect on MHCC-97H cells. Knockdown of p65 blocked MMP-9 activation and enhanced TPL-induced cell apoptosis and survival inhibition, and TPL inhibition of migration and invasion in vitro. CONCLUSIONS TPL treatment inhibited MHCC-97H cell growth, invasion, and metastasis in vitro and vivo, suggesting that TPL could be developed as a potential therapeutic agent for the treatment of HCC.
我们研究了植物来源的试剂雷公藤红素（TPL）是否能有效抑制人肝癌（HCC）细胞的生长和侵袭。
用不同浓度的 TPL 处理 MHCC-97H 细胞不同时间。为了检测 NF-κB 对 TPL 诱导信号通路的影响，MHCC-97H 细胞用 p65 siRNA 或 p65 cDNA 转染，然后用 TPL 处理。我们通过 MTT、软琼脂集落形成试验、流式细胞术和 TUNEL 试验检测细胞存活和凋亡。通过 Matrigel 侵袭和划痕愈合试验测定细胞迁移和侵袭。通过电泳迁移率变动分析（EMSA）检测 NF-κB 活性；通过 ELISA 检测 MMP-9 活性。Western blot 和实时 PCR（RT-PCR）试验检测 p65 和 MMP-9 蛋白和 mRNA 表达。建立 MHCC-97H 细胞裸鼠皮下移植瘤模型，评估 TPL 在体内对肿瘤发生的影响。
我们发现 TPL 处理在体外显著抑制 MHCC-97H 细胞的生长，并呈剂量和时间依赖性诱导细胞凋亡。此外，TPL 处理抑制体外侵袭，并抑制 MHCC-97H 细胞的体内生长和肺转移。TPL 处理使 NF-κB 和 MMP-9 失活。p65 的过表达恢复了 MMP-9 的活性，并抑制了 TPL 对 MHCC-97H 细胞的抗肿瘤作用。p65 的敲低阻断了 MMP-9 的激活，并增强了 TPL 诱导的细胞凋亡和存活抑制，以及 TPL 抑制体外迁移和侵袭。
TPL 处理在体外和体内抑制 MHCC-97H 细胞的生长、侵袭和转移，提示 TPL 可开发为治疗 HCC 的潜在治疗剂。
