ABSTRACT
BACKGROUND: The biological characteristics of hepatic alveolar echinococcosis are similar to cancer lesions. Its biological characteristics of invasive growth and metastasis increase the difficulty of surgery. The fibrosis of the outer capsule wall can inhibit the growth of echinococcus multilocularis and keep the disease in the quiescent stage. The role of mesenchymal stem cells in the fibrosis of the outer capsule wall of echinococcosis remains unclear. OBJECTIVE: To investigate the effect of alveolar echinococcosis protoscolices on the differentiation of mesenchymal stem cells into fibroblasts. METHODS: Bone marrow mesenchymal stem cells were isolated from femur bone marrow of 4-week-old C57BL/6 mice, and cultured by adherent method. Alveolar echinococcosis protoscolices were extracted from gerbils infected with alveolar echinococcus. The experiment was divided into three groups. The alveolar echinococcosis group was co-cultured with the third generation of bone marrow mesenchymal stem cells and the protocephalus of alveolar echinococcosis protoscolices. The Echinococcus granulosus group was co-cultured with the third generation of bone marrow mesenchymal stem cells and the protocephalus of Echinococcus granulosus protoscolices. The simple control group was cultured with the third generation of bone marrow mesenchymal stem cells. At 1, 3, 5, and 7 days of cultivation, the real-time fluorescent quantitative PCR was used to detect collagen type I, collagen type III, transforming growth factor-beta 1 and Smad7 gene expression. Western blot assay was utilized to determine collagen type I, collagen type III, Smad7 and phosphorylated Smad2/3 protein expression. ELISA was applied to measure supernatant collagen type I and collagen type III contents. RESULTS AND CONCLUSION: (1) Real-time fluorescent quantitative PCR detection displayed that transforming growth factor-beta 1, collagen type I, collagen type III mRNA relative expression levels were significantly lower in the alveolar echinococcosis group than in the Echinococcus granulosus group and simple control group (P < 0.05). Smad7 mRNA relative expression was significantly higher in the alveolar echinococcosis group than in the Echinococcus granulosus group and simple control group (P < 0.05). (2) Western blot assay showed that collagen type I, collagen type III and phosphorylated Smad2/3 protein relative expression levels were significantly lower in the alveolar echinococcosis group than in the Echinococcus granulosus group and simple control group (P < 0.05). Smad7 protein relative expression was significantly higher in the alveolar echinococcosis group than in the Echinococcus granulosus group and simple control group (P < 0.05). (3) ELISA exhibited that supernatant collagen type I and collagen type III contents were significantly lower in the alveolar echinococcosis group than in the Echinococcus granulosus group and simple control group (P < 0.05). (4) Alveolar echinococcosis protoscolices may promote bone marrow mesenchymal stem cells to secrete Smad7, inhibit the collagen type I, collagen type III and transforming growth factor-beta 1 through the transforming growth factor-beta/Smad signaling pathway, thereby inhibiting the fibrosis of bone marrow mesenchymal stem cells.
ABSTRACT
Due to the high mortality and rapid disease progression, ovarian cancer remains one of the most common malignancies threatening the health of women. The present study was conducted to explore the anticancer effects and the underlying mechanisms of poricoic acid A (PAA), the main components of Poria cocos, on ovarian cancer. We investigated the anticancer effects of different concentrations of PAA in the SKOV3 cell line. Cell viability and proliferation were examined by CCK-8 assay. Cellular migration and invasion were assessed by the scratch and Transwell migration assays, respectively. The effect of PPA on cell apoptosis was measured by flow cytometry and caspase-3/8/9 colorimetric assay. Western blot was performed to detect protein level changes related to apoptosis and mTOR signaling pathways. The in vivo anticancer effect of PAA was evaluated using xenograft tumorigenesis model in nude mice. Our results showed that PAA suppressed SKOV3 cellular viability, migration, and invasion in a dosage-dependent manner. Flow cytometry results demonstrated PAA treatment could induce SKOV3 cell apoptosis. In addition, increased ratio of LC3-II/LC3-I (a marker for autophagosome formation) was observed after PAA treatment, as well as inhibition of m-TOR and p70s6k phosphorylation. In nude mice, PAA treatment reduced the xenograft tumor weight by 70% (P<0.05). In conclusion, our data suggested that PAA induced apoptosis and autophagy in ovarian cancer via modulating the mTOR/p70s6k signaling axis.