Abstract
Purpose: To investigate the effect of tocotrienolic amide on gastric cancer (GC) cell growth and metastasis, and the underlying mechanism of action.Methods: Gastric cancer (GC) cell lines MKN28 and NCI-N87 were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin solution at 37 ˚C in a humidified atmosphere of 5 % CO2 and 95 % air. Cell invasion and migration were determined using Transwell and wound healing assays, respectively. Realtime quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used for the determination of changes in the levels of expression of Snail, E-cadherin, vimentin, and microRNA-195-5p (miR-195-5p). In vivo tumor growth inhibition was determined 45 days after establishment of GC xenografts in nude mice.Results: Treatment of MKN28 and NCI-N87 cells with tocotrienolic amide significantly and dosedependently reduced their invasiveness and migratory capacity (p < 0.05). It also significantly and dosedependently downregulated the mRNA and protein expressions of Snail and vimentin, but significantly upregulated E-cadherin expression (p < 0.05). The mRNA expression of miR-195-5p was significantly and dose-dependently upregulated in MKN28 and NCI-N87 cells treated with tocotrienolic amide, but was downregulated after transfection with miR-195-5p inhibitor (p < 0.05). Transfection of MKN28 and NCI-N87 cells with miR-195-5p inhibitor significantly and dose-dependently upregulated mRNA and protein expressions of Snail (p < 0.05). Moreover, treatment of GC mice with TCTA led to significant and dose-dependent reduction in tumor weight and volume (p < 0.05).Conclusion: These results suggest that tocotrienolic amide inhibits the growth and metastasis of GC cells by directly targeting Snail and vimentin genes, and thus can potentially be developed for the management of gastric cancer.
 Keywords: Gastric cancer, Metastasis, Protein expression, Tocotrienolic amide, Tumor volume
Highlights
Gastric cancer (GC), a malignant tumor that develops in the inner lining of the stomach, is the fifth most common cancer worldwide
The mRNA expression of miR-195-5p was significantly and dose-dependently upregulated in MKN28 and NCI-N87 cells treated with tocotrienolic amide, but was downregulated by transfection with miR-195-5p inhibitor (p < 0.05; Figure 3)
This study investigated the effect of tocotrienolic amide on GC cell growth and metastasis, and the mechanism involved
Summary
Gastric cancer (GC), a malignant tumor that develops in the inner lining of the stomach, is the fifth most common cancer worldwide. The present study investigated the inhibitory effect of tocotrienolic amide on the migration and invasion of gastric cancer cells. Gastric cancer cell lines (MKN28 and NCI-N87) were obtained from Biochemistry and Cell Biology Institute, Shanghai, China. Gastric cancer cells (MKN28 and NCI-N87) in logarithmic growth phase were seeded in 24-well plates at a density of 1 × 106 cells/mL and cultured in DMEM for 24 h. Light Cycler 1536 RT-PCR detection system was used for the measurement of the expression levels of Snail, E-cadherin, vimentin, and miR-195-5p. Gastric cancer (GC) cells were cultured in serum-free medium with equal volume of miR195-5p inhibitor or control, each at concentration of 10 μmol/L. The tumors formed in nude mice were excised and weighed to determine the effect of tocotrienolic amide on tumor growth in the treatment groups.
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