ABSTRACT
OBJECTIVE: To explore the effect of micro-ribonucleic acid (miR)-187 on cisplatin (DDP) resistance of gastric cancer cells by regulating the transforming growth factor-ß (TGF-ß)/Smad signaling pathway. MATERIALS AND METHODS: DDP-sensitivities in GES-1, SGC7901, and SGC7901/DDP cells were detected via Cell Counting Kit-8 (CCK-8) assay. The differential expression of miR-187 of these cell lines was detected by Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR). DDP-resistant gastric cancer cells SGC7901/DDP were divided into control group (blank control), miR-187 inhibitor group (SGC7901/DDP cells transfected with miR-187 inhibitor), and miR-187 mimic group (SGC7901/DDP cells transfected with miR-187 mimic). The protein expressions of miR-187, TGF-ß1, p-Smad4, excision repair cross-complementation group 3 (ERCC3), and ERCC4 were determined through RT-qPCR, immunohistochemistry, and Western blotting. The apoptosis in each group was detected by flow cytometry. RESULTS: MiR-187 level had a negative correlation with DDP-resistance of GES-1, SGC7901, and SGC7901/DDP cells, and among them, the GES-1 cells had the lowest DDP-resistance and the highest expression of miR-187. CCK-8 assay revealed that compared with that in the control group, DDP-resistance significantly declined in the miR-187 mimic group, while it was significantly enhanced in miR-187 inhibitor group (p<0.01). According to the results of flow cytometry, after treatment with 100 nM DDP for 12 h, the apoptotic rate in miR-187 mimic group enhanced, while it was markedly reduced in the miR-187 inhibitor group (p<0.01). Western blotting and immunohistochemistry results showed that expressions of TGF-ß1 and p-Smad4 were significantly downregulated in the miR-187 mimic group, while they were upregulated in the miR-187 inhibitor group (p<0.01). Besides, compared with the control group, ERCC3 and ERCC4 were downregulated in the miR-187 mimic group, while upregulated in miR-187 inhibitor group (p<0.01). CONCLUSIONS: The overexpression of miR-187 alleviates DDP-resistance in gastric cancer cells by inhibiting the TGF-ß/Smad signaling pathway.
Subject(s)
Cisplatin/pharmacology , Drug Resistance, Neoplasm , MicroRNAs/genetics , Signal Transduction , Stomach Neoplasms/genetics , Cell Line, Tumor , Down-Regulation , Gene Expression Regulation, Neoplastic , Humans , Phosphorylation , Smad Proteins/metabolism , Stomach Neoplasms/drug therapy , Stomach Neoplasms/metabolism , Transforming Growth Factor beta/metabolismABSTRACT
A recombinant fowlpox virus vaccine expressing key protective Mycoplasma gallisepticum antigens could facilitate in the prevention both of fowlpox virus and M. gallisepticum infections. Vectormune FP-MG vaccine, a recombinant fowlpox virus expressing both M. gallisepticum 40k and mgc genes, was assessed for its safety in 8-wk-old specific-pathogen-free White Leghorn chickens. The vaccine virus was serially passaged 5 times by wing-web inoculation. Based on the postinoculation clinical observation, gross pathological examination of air sacs and peritoneum, genetic stability evaluation, virus shedding and tissue distribution detection, horizontal transmission ability determination, and protection against fowlpox virus challenge, the Vectormune FP-MG vaccine possesses a high level of safety.
