Background

The therapeutic efficiency of Traditional Chinese Medicine (TCM) in suppressing the recurrence and metastasis of hepatocellular carcinoma (HCC) has been well proved.

Objective

The aim of this study is to investigate the role of Bie Jia Jian pill (BJJP) combined with bone mesenchymal stem cells (BMSCs) in HCC progression.

Methods

Flow cytometry was used to identify BMSCs isolated from BALB/c mice. The expressions of biomarkers and apoptosis rate of cancer stem cells (CSCs) enriched from Huh7 cells were also measured. The osteogenic differentiation and adipogenic differentiation ability of isolated BMSCs was determined by oil red O staining and Alizarin Red Staining. CSCs were used to establish the orthotopic HCC model. Histological changes in the liver tissues were examined by hematoxylin–eosin (H&E) staining and Van Gieson (VG) staining. The cell apoptotic rate in the cancer tissues was detected by TUNEL staining. The cell proliferation antigen Ki67 in the cancer tissues were detected by immunofluorescence assay and PCR, respectively. The levels of CSCs cellular surface markers (CD24, CD133 and EpCAM) and Wnt/β-catenin signal pathway related proteins were detected by PCR and western blot.

Results

Treatment of BJJP or BMSCs both improved the morphology induced by HCC and suppressed the differentiation ability of CSCs, as evidenced by down-regulated expressions of CD24, CD133, EpCAM and Ki67. The protective effect of BJJP or BMSCs in cancer tissues can be enhanced by the combination of BJJP and BMSCs. In addition to that, BJJP or BMSCs alone was found to increase the expression of miR-140 and promote cell apoptosis in CSCs, while down-regulation of miR-140 partially reversed the protective effect of BMSCs or BJJP + BMSCs on cancer tissues. BJJP + BMSCs treatment together also can down-regulate the expressions of Wnt3a and β-catenin.

Conclusions

These results proved the inhibitory role of BJJP + BMSCs in HCC development through regulating miR-140 and Wnt/β-catenin signal pathway.

中文翻译：

---

鳖甲煎丸增强骨髓间充质干细胞对肝癌进展的改善作用

背景

中医药在抑制肝细胞癌（HCC）复发转移方面的疗效已得到充分证明。

客观的

本研究旨在探讨鳖甲煎丸 (BJJP) 联合骨髓间充质干细胞 (BMSCs) 在 HCC 进展中的作用。

方法

流式细胞术用于鉴定从 BALB/c 小鼠分离的 BMSC。还测量了从 Huh7 细胞中富集的癌症干细胞 (CSC) 的生物标志物表达和凋亡率。通过油红O染色和茜素红染色测定分离的BMSCs的成骨分化和成脂分化能力。CSC 用于建立原位 HCC 模型。通过苏木精-伊红 (H&E) 染色和 Van Gieson (VG) 染色检查肝组织的组织学变化。TUNEL染色检测癌组织中细胞凋亡率。分别采用免疫荧光法和PCR检测癌组织中的细胞增殖抗原Ki67。CSCs细胞表面标志物（CD24、

结果

BJJP 或 BMSCs 的治疗既改善了 HCC 诱导的形态，又抑制了 CSCs 的分化能力，CD24、CD133、EpCAM 和 Ki67 的表达下调证明了这一点。BJJP和BMSCs的结合可以增强BJJP或BMSCs对癌组织的保护作用。除此之外，发现BJJP或BMSCs单独增加miR-140的表达并促进CSCs中的细胞凋亡，而miR-140的下调部分逆转了BMSCs或BJJP + BMSCs对癌组织的保护作用。BJJP+BMSCs联合治疗也可以下调Wnt3a和β-catenin的表达。

结论

这些结果证明了BJJP+BMSCs通过调控miR-140和Wnt/β-catenin信号通路在HCC发展中的抑制作用。