ABSTRACT
ObjectiveTo explore the inhibitory effect of water extract of Broussonetiae Fructus on hepatocellular carcinoma （HCC） induced by diethyl nitrosamine （DEN） in mice based on homologous phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B （PTEN/PI3K/Akt） signaling pathway. MethodThe primary HCC mouse model was constructed by intraperitoneal injection of DEN solution， and the HCC mice were randomly divided into model group， sorafenib group （0.01 g·kg-1·d-1）， low-dose Broussonetiae Fructus water extract group （0.9 g·kg-1·d-1）， medium-dose Broussonetiae Fructus water extract group （1.8 g·kg-1·d-1）， and high-dose Broussonetiae Fructus water extract group （3.6 g·kg-1·d-1）， with 10 mice in each group. Another 10 C57BL/6 mice were selected as a control group and intraperitoneally injected with an equal volume of normal saline. Mice were treated with different concentrations of Broussonetiae Fructus water extract when liver cancer-like white nodules appeared. sorafenib group was treated with sorafenib. The control group and model group were intraperitoneally injected with normal saline. The activities of alkaline phosphatase （ALP）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， and γ-glutamyl transferase （γ-GT） in the serum of mice were detected by the biochemical analyzer. The expression levels of alpha-fetoprotein （AFP） and carcinoembryonic antigen （CEA） were detected by enzyme-linked immunosorbent assay （ELISA）. The degree of hepatocyte canceration and hepatocyte injury were observed by Hematoxylin-eosin （HE） and Masson staining. The proliferation of HCC cells was observed by immunohistochemical staining. The apoptosis of HCC cells in mice was observed by erminal-deoxynucleotidyl transferase mediated nick end labelling （TUNEL） staining. The expression levels of PTEN， PI3K， Akt， and p-Akt proteins related to the PTEN/PI3K/Akt signaling pathway were detected by Western blot. ResultCompared with the control group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the model group were significantly increased （P<0.01）. Carcinogenesis and inflammatory cell infiltration were obvious in liver tissue of mice， and a large number of blue collagen fiber hyperplasia was found. The number of Ki67 positive cells was significantly increased （P<0.01）， and the expression level of PTEN protein was significantly decreased， while PI3K and p-Akt protein expression was increased （P<0.01）. Compared with the model group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the medium-dose and high-dose Broussonetiae Fructus water extract groups were significantly decreased （P<0.05， P<0.01）. The degree of carcinogenesis and inflammatory cell infiltration in liver tissue were reduced， and the collagen fiber hyperplasia was significantly reduced. The number of Ki67 positive cells was significantly decreased， and the number of TUNEL positive apoptotic cells was significantly increased （P<0.05， P<0.01）. PTEN protein expression was increased， while p-Akt protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionThe water extract of Broussonetiae Fructus has a significant inhibitory effect on DEN-induced primary HCC in mice， and its mechanism may be related to the regulation of key protein expressions in the PTEN/PI3K/Akt signaling pathway.
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AIM: To investigate the sensitization of flavonoids from Tetrastigma hemsleyanum (FTH) on gefitinib (GEF)-resistant lung adenocarcinoma cells. METHODS: The viabilities of A549 and A549/GR cells treated with FTH and GEF were detected by MTT method. The apoptotic rates and cell cycles of A549/GR cells treated with FTH and GEF were detected by Flow cytometry. The anti-tumor effects of flavonoids from FTH and GEF were assayed in A549/GR tumor-bearing mice. The expressions of proteins (PTEN, PI3K, p-PI3K, AKT, p-AKT) were detected by Western blot analysis. RESULTS: Compared with GEF group, FTH significantly enhanced the inhibition of GEF on the proliferation of A549/GR cells (P< 0.05). Combination with FTH and GEF significantly increased the apoptosis of A549/GR cells which were arrested at the G
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BACKGROUND: Acute myeloid leukemia (AML) is an aggressive and mostly incurable hematological malignancy with frequent relapses after an initial response to standard chemotherapy. Therefore, novel therapies are urgently required to improve AML clinical outcomes. 4-Amino-2-trifluoromethyl-phenyl retinate (ATPR), a novel all-trans retinoic acid (ATRA) derivative designed and synthesized by our team, has been proven to show biological anti-tumor characteristics in our previous studies. However, its potential effect on leukemia remains unknown. The present research aims to investigate the underlying mechanism of treating leukemia with ATPR in vitro. METHODS: In this study, the AML cell lines NB4 and THP-1 were treated with ATPR. Cell proliferation was analyzed by the CCK-8 assay. Flow cytometry was used to measure the cell cycle distribution and cell differentiation. The expression levels of cell cycle and differentiation-related proteins were detected by western blotting and immunofluorescence staining. The NBT reduction assay was used to detect cell differentiation. RESULTS: ATPR inhibited cell proliferation, induced cell differentiation and arrested the cell cycle at the G0/G1 phase. Moreover, ATPR treatment induced a time-dependent release of reactive oxygen species (ROS). Additionally, the PTEN/PI3K/Akt pathway was downregulated 24 h after ATPR treatment, which might account for the anti-AML effects of ATPR that result from the ROS-mediated regulation of the PTEN/PI3K/AKT signaling pathway. CONCLUSIONS: Our observations could help to develop new drugs targeting the ROS/PTEN/PI3K/Akt pathway for the treatment of AML.
Subject(s)
Humans , Retinoids/pharmacology , Reactive Oxygen Species/metabolism , Antineoplastic Agents/pharmacology , Fluoroimmunoassay , Leukemia, Myeloid, Acute , Signal Transduction , Down-Regulation , Cell Differentiation/drug effects , Cell Survival/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , PTEN Phosphohydrolase/drug effects , PTEN Phosphohydrolase/metabolism , Proto-Oncogene Proteins c-akt/drug effects , Proto-Oncogene Proteins c-akt/metabolismABSTRACT
@#Objective:To explore the mechanism of EYA1 (eyes absent 1) inhibiting the malignant progression of gastric cancer SGC7901 cells through regulating PTEN/PI3K/AKT signaling pathway. Methods: Twenty-nine pairs of gastric cancer tissues and para-cancerous tissues collected at the General Surgery center, Southwest Hospital Affiliated to Military Medical University during June 2016 and June 2018 were used in this study. Wb and RT-PCR assays were used to test the mRNA and protein expressions of EYA1 in gastric cancer tissues and the paired para-cancerous tissues; Transfection with plasmid or siRNAs were used to up-regulate or down-regulate EYA1 or PTEN expression in gastric cancer SGC-7901 cells; MTT, Flow Cytometry, Wound Healing and Transwell assays were carried out to detect cell proliferation, apoptosis, metastasis and invasion abilities, respectively. Results: EYA1 expression was decreased in gastric cancer tissues as compared with the para-cancerous tissues at both mRNA and protein levels (P<0.01); EYA1 over-expression significantly enhanced the proliferation, metastasis and invasion of SGC-7901 cells (all P<0.05), and inhibited cell apoptosis (P<0.05); moreover, its over-expressionsignificantly increased the expression of PTEN, and inhibited the activation of PI3K/AKT pathway (all P< 0.05 or P<0.01). However, the above effects mediated by EYA1 up-regulation were significantly impaired after the knockout of PTEN (all P<0.05 or P<0.01). Conclusion: EYA1 can inhibit the malignant progression of gastric cancer SGC-7901 cells through promoting the expression of PTEN and activating PI3K/AKT pathway.