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This article has been cited by other articles in PMC. Abstract Background Lung cancer is the most common malignant tumor worldwide. Ganoderan GDN is one of the components of Ganoderma lucidum polysaccharides.
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Methods Cell growth was measured by Cell Counting kit-8 and colony formation assay, while cell motility was measured by transwell assay and wound healing assay. Apoptosis was measured by flow cytometry analysis and TUNEL staining, and protein expression was detected by Western prostate specific tumor marker and immunohistochemistry.
Results Previous studies have shown that GDNB has the effects of hyperglycemic and kidney protection. However, the role of GDNB in tumors is currently unknown. Besides, transwell assay and wound healing assay showed that GDNB inhibited invasion and migration of HA and A cells in a concentration-dependent manner.
Moreover, Western blotting also showed that GDNB downregulated the levels of N-cadherin, vimentin and Snail in HA and A cells in a dose-dependent manner, while it upregulated the level of E-cadherin. Animal experiments revealed that GDNB inhibited tumor growth and metastasis, and induced apoptosis of tumor cells in vivo.
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Conclusion Collectively, all data suggest that GDNB regulates the growth, motility and apoptosis of non-small cell lung cancer cells through ERK signaling pathway in vitro and in vivo. Ganoderma lucidum is one of the famous Chinese herbal medicines in China and has a history of more than 2, years.
Extracellular signal-regulated protein kinase ERK signaling pathway is a classical mitogen-activated protein kinases MAPKs signal transduction pathway and plays an important role in cell proliferation, 17 invasion, migration, 18 differentiation and apoptosis. In the current study, we explored the role of GDNB in lung cancer and its underlying molecular mechanisms.
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Our results indicate that GDNB can significantly inhibit the growth and motility of lung cancer cells, and induce cell apoptosis by inactivating the Prostate specific tumor marker leech prostatitis pathway in vitro and in vivo.
Our findings reveal that GDNB may be a potential anticancer drug in the treatment of lung cancer. GDNB was bought from Hubei jusheng technology co. Furtherly, the cells were exposed to 10 µl CCK-8 solution at 37°C for 1 hrs.
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Subsequently, HA and A cells were subjected to different concentrations 0. The cells were cultured for two weeks to form colonies.
After that, the cells were stained with crystal violet 0. Each assay was performed in triplicate. Thereafter, the cells 4.
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Then, μl of complete medium was added to the basal side chamber. After incubation at 37°C for 24 hrs, the cells were fixed with ice-cold methanol for 5 min at room temperature and stained with 0. The remaining cells on the upper chambers were gently removed with a wet cotton swab, and the cells in the low chambers were stained with Hematoxylin dye μl for 10 min and photographed using a microscope Leica Microsystems, Wetzlar, Germany with five randomly selected image fields.
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Wound Healing Assay Migration ability was detected using the Wound healing assay. Subsequently, monolayer cells were cut with a sterile μl pipette tip to create an artificial wound.
Thereafter, cells were cultured under serum-starved conditions for 24 hrs. After that, the cells were washed with PBS for three times to remove the fallen cells. The cell migration was observed with a phase contrast microscope, and images were taken with a D camera Canon, Japan.
HA and A cells were seeded in 6-well plates 1. Thereafter, A cells 1. After successful modeling, lung tumor-bearing mice were randomly divided into 4 prostate specific tumor marker ten in each group : Control group The mice were intragastrically administered with 0.
After eight weeks later, the mice were sacrificed, and tumors were taken out, weight and photograph.
Then, the tumor tissues were cut into pieces and stored for subsequent research. In brief, the tumor tissue embedded in paraffin was cut into 5 μm-thickness prostate specific tumor marker.
Then, the sections were washed with PBS for three times and permeable with proteinase K at 37°C for 20 min. Whereafter, the labeled DNA was observed using a peroxidase-conjugated anti-fluorescein antibody.
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The percentage of tunel positive cells to the total number of cells is considered to be the apoptotic rate. Immunohistochemistry IHC assay was employed to detect the expression of caspase 3, Ki67 and vimentin. Then, H2O2 solution was added to block endogenous peroxidase activity.
Finally, the samples were co-cultured with 3,3ʹ-diaminobenzidine and observed under a light microscope Nikon. Statistical Analysis Data are expressed as mean ± standard deviation SD of at least three independent experiments. GDNB treatment for 24 hrs and 48 hrs had no significant effect on the viability of WI cells, but prostate specific tumor marker decreased the viability of A and HA cells in a concentration-dependent manner. However, when the dose of GDNB exceeded 1.
Therefore, prostate specific tumor marker subsequent experiments in this study used a treatment time of 48 hrs. Next, we explored the effects of different concentrations of GDNB 0. All these data indicate that GDNB inhibits the growth of non-small cell lung cancer cells.