Inhibitory Effect of Cardamonin on Human Osteosarcoma Cells and Its Underlying Mechanism / 中国药学杂志

OBJECTIVEP: To analyze and confirm the inhibitory effect of cardamonin(CAR) on human osteosarcoma(OS) cells and its related mechanism. METHODS: Human osteosarcoma cells were treated with 0, 4, 12, 16 μmol•L-1 CAR and 0.08% DMSO, respectively. Cell proliferation was detected by crystal violet staining and MTT assay. Cell apoptosis was detected by Hoechst staining and flow cytometry(FCM). Cell migration ability was detected by scratch healing test. Cell invasion ability was detected by Transwell method. Western blot detects changes in cell proliferation, apoptosis, migration and invasion-associated proteins and Wnt/β-catenin signaling pathway-related proteins. RESULTS: CAR inhibits proliferation, migration and invasion of osteosarcoma cells. CAR inhibits the expression of PCNA, MMP-7 and vimentin. CAR inhibits the expression of Bcl-2, promotes the expression of apoptotic markers caspase 3 and cleaved-caspase 3. CAR inhibits the expression of the key molecule β-catenin of Wnt/β-catenin signaling and its downstream target molecules cyclin D1 and c-Myc. CONCLUSION: CAR can inhibit the proliferation, invasion and migration of osteosarcoma cells, but promote its apoptosis. Its molecular mechanism may be related to the interference of Wnt/beta-catenin signaling pathway activation.

Microrna-27a knockdown inhibits proliferation and metastasis in renal cell carcinoma through wnt/p-catenin signaling pathway / 中国药理学通报

Aim To explore the role of miR-27a on the proliferation and metastasis of renal cell carcinoma (RCC) and its mechanism. Methods The expression of miR-27a in RCC cancer tissues, para-carcinoma tissues, RCC cells (Caki-1, 786-0 and ACHN) and normal renal tubular epithelial cells ( HK2) were detected by RT-qPCR. After miR-27a-inhibitor transfect-ed into 786-0 and ACHN cells, cell proliferation was measured by CCK-8 assay; cell colony formation was detected by colony formation assay; cell migration and invasion were detected by cell wound healing assay and Transwell assay, respectively; the pretein expression of P-catenin was detected by Western blot. After trans-fected miR-27a inhibitor into ACHN cells then treated with LiCl (Wnt/p-catenin signal agonist), cell proliferation , migration and invasion were detected. Results The expression of miR-27a in RCC cancer tissues was significantly higher than that in para-carcinoma tissues, and increased with stage progression. Compared with HK-2 cells, the expression levels of miR-27a in RCC cells were elevated. After transfection with miR-27a inhibitor, the cell colony formation, cell prolifera-tion, invasion and migration ability of 786-0 and ACHN cells were significantly reduced. MiR-27a in-i hibitor reduced the expression of p-catenin in ACHN cells. LiCl promoted the proliferation, invasion and migration ability of ACHN cells transfected with miR-27a inhibitor. Conclusions MiR-27a is highly expressed in RCC cancer tissues and RCC cells, and knockdown of miR-27a inhibits proliferation and metastasis in RCC cells through Wnt/p-catenin signaling pathway.

Polydatin inhibits cell proliferation, invasion and migration, and induces cell apoptosis in hepatocellular carcinoma

Polydatin, a small molecule from Polygonum cuspidatum, has many biological functions, particularly anti-cancer effects. However, the anti-cancer effects of polydatin in hepatocellular carcinoma (HCC) have not been examined yet. In the present study, MTT assay, BrdU assay, transwell invasion assay, and wound healing assay were performed to determine cell proliferation, invasion and migration. Flow cytometry and TUNEL assay were used to measure cell apoptosis. Quantitative real-time PCR and western blotting assays were used to determine mRNA and protein expression levels. Xenograft experiment was performed to determine the in vivo anti-tumor effect of polydatin. Immunostaining was performed to analyze the expression of caspase-3 and Ki-67. Our results showed that polydatin inhibited cell proliferation in a concentration-dependent and time-dependent manner in the HCC cell lines. Polydatin also induced cell apoptosis in a concentration-dependent manner possibly via increasing the caspase-3 activity, and up-regulating the protein expression of caspase-3, caspase-9, Bax, and down-regulating the protein expression of Bcl-2. In addition, polydatin treatment had an inhibitory effect on cell proliferation, invasion and migration in HCC cell lines. Polydatin treatment also suppressed the Wnt/beta-catenin signaling activities in HCC cells. Polydatin treatment significantly reduced tumor growth in nude mice inoculated with HepG2 cells, suppressed the expression of Ki-67, and increased caspase-3 expression and TUNEL activity. Our data indicated the important role of polydatin for the suppression of HCC progression.

Opposite Interplay Between the Canonical WNT/β-Catenin Pathway and PPAR Gamma: A Potential Therapeutic Target in Gliomas / 神经科学通报·英文版

In gliomas, the canonical Wingless/Int (WNT)/β-catenin pathway is increased while peroxisome proliferator-activated receptor gamma (PPAR-γ) is downregulated. The two systems act in an opposite manner. This review focuses on the interplay between WNT/β-catenin signaling and PPAR-γ and their metabolic implications as potential therapeutic target in gliomas. Activation of the WNT/β-catenin pathway stimulates the transcription of genes involved in proliferation, invasion, nucleotide synthesis, tumor growth, and angiogenesis. Activation of PPAR-γ agonists inhibits various signaling pathways such as the JAK/STAT, WNT/β-catenin, and PI3K/Akt pathways, which reduces tumor growth, cell proliferation, cell invasiveness, and angiogenesis. Nonsteroidal anti-inflammatory drugs, curcumin, antipsychotic drugs, adiponectin, and sulforaphane downregulate the WNT/β-catenin pathway through the upregulation of PPAR-γ and thus appear to provide an interesting therapeutic approach for gliomas. Temozolomide (TMZ) is an antiangiogenic agent. The downstream action of this opposite interplay may explain the TMZ-resistance often reported in gliomas.

Atractylochromene Is a Repressor of Wnt/beta-Catenin Signaling in Colon Cancer Cells

Wnt/beta-catenin signaling pathway was mutated in about 90% of the sporadic and hereditary colorectal cancers. The abnormally activated beta-catenin increases the cancer cell proliferation, differentiation and metastasis through increasing the expression of its oncogenic target genes. In this study, we identified an inhibitor of beta-catenin dependent Wnt pathway from rhizomes of Atractylodes macrocephala Koidzumi (Compositae). The active compound was purified by activity-guided purification and the structure was identified as 2,8-dimethyl-6-hydroxy-2-(4-methyl-3-pentenyl)-2H-chromene (atractylochromene, AC). AC suppressed beta-catenin/T-cell factor transcriptional activity of HEK-293 reporter cells when they were stimulated by Wnt3a or inhibitor of glycogen synthase kinase-3beta. AC down-regulated the nuclear level of beta-catenin through the suppression of galectin-3 mediated nuclear translocation of beta-catenin in SW-480 colon cancer cells. Furthermore, AC inhibits proliferation of colon cancer cell. Taken together, AC from A. macrocephala might be a potential chemotherapeutic agent for the prevention and treatment of human colon cancer.

Mest Attenuates CCl4-Induced Liver Fibrosis in Rats by Inhibiting the Wnt/beta-Catenin Signaling Pathway

BACKGROUND/AIMS: The Wnt/beta-catenin signaling pathway has been reported to play an important role in liver fibrosis. This study was designed to investigate whether mesoderm-specific transcript homologue (Mest), a strong negative regulator of Wnt/beta-catenin signaling, could inhibit liver fibrosis. METHODS: pcDNA-Mest was transfected into hepatic stellate cells (HSCs) and rats. Rats were randomly divided into four groups: normal group (normal saline), treatment group (pcDNA-Mest+CCl4), control group (pcDNA-neo+CCl4), and model group (normal saline+CCl4). Changes in liver pathology were evaluated by hematoxylin and eosin and Masson's trichrome staining. The levels of alanine transaminase, aspartate transaminase, lactic dehygrogenase, hyaluronic acid, and laminin in the serum and hydroxyproline in the liver were detected by biochemical examination and radioimmunoassay, respectively. The expression and distribution of beta-catenin, alpha-smooth muscle actin (alpha-SMA), Smad3, and tissue inhibitor of metalloproteinase type I were determined, and the viability of the HSCs was tested. RESULTS: Our data demonstrate that Mest alleviated CCl4-induced collagen deposition in liver tissue and improved the condition of the liver in rats. Mest also significantly reduced the expression and distribution of beta-catenin, alpha-SMA and Smad3 both in vivo and in vitro, in addition to the viability of HSCs in vitro. CONCLUSIONS: We found that Mest attenuates liver fibrosis by repressing beta-catenin expression, which provides a new therapeutic approach for treating liver fibrosis.

Affection of Wnt/beta-catenin in Titanium Particles Challenged Osteoblasts / 대한정형외과연구학회지

PURPOSE: The intracellular mechanisms that lead to periprosthetic osteolysis including impaired bone forming activity of osteoblast remain incompletely characterized. To determine the possibility that Ti-particles play a role to regulate Wnt/beta-catenin signaling pathway in impaired osteogenesis, we analyzed the stability of beta-catenin and the transcriptional changes of regulators for Wnt/beta-catenin signaling pathway in MC3T3-E1 osteoblast cells. MATERIALS AND METHODS: Ti-particles were prepared by sterilizing and counted on the microscopy. Transcriptional changes of OPG, RANKL, LRP5, LRP6, DKK1 and sFRP2 were determined by real-time RTPCR. Protein level of beta-catenin and GSK3beta was detected using Western blotting and immunofluorescence staining. RESULTS: After 4 hours of treatment of Ti-particles, OPG/RANKL mRNA ratio was significantly decreased. And also, decreased protein levels of beta-catenin and phospho-GSK3beta were detected. Using immunofluorescence stain, it was confirmed that Ti-particles suppressed nucleus staining of beta-catenin induced by Wnt3a conditioned medium. The results of real-time RT-PCR showed reduced level of LRP5 and LRP6 transcripts, and induced level of DKK1 and sFRP2 transcripts by challenging of Ti-particles CONCLUSION: Our report suggests that Ti-particles may play a crucial role in the regulation of Wnt/beta-catenin signaling pathway in osteoblast through the transcriptional changes of membrane receptors and extracellular inhibitors for Wnt.