Abstract
BackgroundBased on clinical medications and related studies, we established a Yang-Gan Jie-Du Sang-Jie (YGJDSJ) herbal formula for hepatocarcinoma treatment. In present study, we evaluated the anti-cancer potential of YGJDSJ on suspension-grown human hepatocellular carcinoma Bel-7402 cells.MethodsBel-7402 cells were cultured in poly(2-hydroxyethyl methacrylate) (poly-HEMA) coated plates and treated with YGJDSJ. Anchorage-independent cell growth was detected by cell Counting Kit-8 (CCK-8) assay and soft agar colony formation assay. Anoikis was detected by ethdium homodimer-1 (EthD-1) staining and flow cytometry analysis. Caspases activities were detected by the cleavage of chromogenic substrate. Reactive oxygen species (ROS) was detected by 2′,7′-dichlorofluorescin diacetate (DCFH-DA) staining. Protein expression and phosphorylation was identified by western blot. Protein expression was knocked-down by siRNA.ResultsYGJDSJ inhibited the proliferation of Bel-7402 cells in poly-HEMA coated plates and anchorage-independent growth of Bel-7402 cells in soft agar. YGJDSJ also induced anoikis in Bel-7402 cells as indicated by EthD-1 staining and flow cytometry analysis. YGJDSJ activated caspase-3, − 8, and − 9 in suspension-grown Bel-7402 cells. The pan-caspase inhibitor Z-VAD-FMK significantly abrogated the effects of YGJDSJ on anoikis in suspension-grown Bel-7402 cells. In addition, YGJDSJ increased ROS in suspension-grown Bel-7402 cells. The ROS scavenger N-acetyl-L-cysteine (NAC) partially attenuated YGJDSJ-induced activation of caspase-3, − 8 and − 9 and anoikis in suspension-grown Bel-7402 cells. Furthermore, YGJDSJ inhibited expression and phosphorylation of protein tyrosine kinase 2 (PTK2) in suspension-grown Bel-7402 cells. Over-expression of PTK2 significantly abrogated YGJDSJ induced anoikis.ConclusionsYGJDSJ inhibits anchorage-independent growth and induce caspase-mediated anoikis in Bel-7402 cells, and may relate to ROS generation and PTK2 downregulation.
Highlights
Based on clinical medications and related studies, we established a Yang-Gan Jie-Du Sang-Jie (YGJDSJ) herbal formula for hepatocarcinoma treatment
YGJDSJ inhibits anchorage-independent growth of Bel-7402 cells Poly-HEMA-coated culture plates were used to observe the effects of YGJDSJ on anchorage-independent growth in Bel-7402 cells
The results show that YGJDSJ inhibited proliferation of Bel-7402 cells in the poly-HEMA coated plates in a dose-dependent manner (P < 0.01) (Fig. 1a)
Summary
Based on clinical medications and related studies, we established a Yang-Gan Jie-Du Sang-Jie (YGJDSJ) herbal formula for hepatocarcinoma treatment. We evaluated the anti-cancer potential of YGJDSJ on suspension-grown human hepatocellular carcinoma Bel-7402 cells. Hepatocarcinoma is one of the most common malignancies worldwide, and ranks second and sixth as the cause of cancer deaths in men and women, respectively [1]. The treatment options for hepatocarcinoma mostly include surgery, transhepatic artery chemoembolization (TACE), and targeted therapy. Surgery, comprising either hepatectomy and/or liver transplantation, is the only treatment that can possibly cure hepatocarcinoma. Only patients with early-stage hepatocarcinoma are eligible for curative surgery. Hepatocarcinomas tend to metastasize to the lungs, bones, adrenal glands, and other distant organs via blood circulation. Hepatocarcinomas can metastasize to the lymph nodes via circulating lymph, including portal
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