RESUMO
INTRODUCTION: The incidence of colorectal cancer is steadily increasing, and the detection of related molecular targets is critical for its diagnosis and treatment. Long noncoding RNA (lncRNA) can play a regulatory role before and after genome transcription, and epigenetic regulation is involved in the process of tumorigenesis and tumor development. METHODS: In this study, qRT-PCR was performed to evaluate the expression of AK093407 in colon cancer and colon para-carcinoma tissues and HCT-15 and HCT-116 cells. SiRNA was transfected into HCT-15 and HCT-116 cells to knock down lncRNA-AK093407. Then, MTT assay was used to test cell proliferation, and flow cytometry was used to test apoptosis and cell cycle. The protein expression of caspase-3, caspase-8, caspase-9, bax, bcl-2, cyclin-A1, cyclin-B1, cyclin-D1, cyclin- E1, p21, p27, and p-Stat3 was determined by Western blot. RESULTS: The results showed that the expression of AK093407 in human colon cancer tissue was higher than in para-carcinoma tissue. The amount of AK093407 in HCT-15 and HCT-116 cells was higher than that in normal colorectal epithelial NM460 cells. When AK093407 was silenced, the proliferation of HCT-15 and HCT-116 cells decreased, the apoptosis rate increased, the cell cycle was arrested in the G1/S phase, the expression of caspase-3, caspase-8, caspase-9, bax, cyclin-A1, cyclin- B1, p21, p27 increased, and the expression of bcl-2, cyclin-D1, cyclin-E1, p-Stat3 decreased. CONCLUSION: These results showed that knockdown of AK093407 could inhibit colon cancer cell proliferation, induce apoptosis and cell cycle arrest, influence the expression of vital factors in mitochondrial apoptosis pathway and cell cycle regulatory pathway, and may negatively regulate JAK/STAT3 through down-regulating p-Stat3.
Assuntos
Neoplasias do Colo , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/genética , Caspase 3 , Caspase 9 , Caspase 8 , Epigênese Genética , Proteína X Associada a bcl-2 , Neoplasias do Colo/genéticaRESUMO
This study was aimed to observe the influence of Discornin Tablets on activation nuclear transcription factor NF-κBp65 of rheumatoid arthritis (RA) cell model as well as the expression of MMP-9, VEGF and tumor necrosis factor-α (TNF-α). Interleukin-17 (IL-17) and TNF-α were used for stimulating RSC-364 cells. Discornin Tablets at different concentrations were used for intervention. The influence of Discornin Tablets in different concentrations on cell viability was detected by MTT method. Expressions of NF-κBp65 and its inhibitory protein (IκB-α) in each group were detected by western blot method. Changes in VEGF, MMP-9 and TNF-α protein levels in cell broth supernatant were checked by ELISA. The results showed that Discornin Tablets can promote the expression of κB inhibitory pro-tein, reduce the high expression of NF-κB protein level, and inhibit the cellular secretion of VEGF, MMP-9 and TNF-α. It was concluded that Discornin Tablets had negative regulation effect on nuclear transcription factor κB of RSC-364 cells. It can increase the expression of IκB-α, as well as reduce the secretion of inflammation factors and blood vessel newborn factors. It suggested that Discornin Tablets may have the potential regulation effect on RA.
RESUMO
This study was aimed to observe the influence of water-solubility nipponica saponin on activation of TNF-α+IL-17-induced rat fibroblast-like synovial cell line RSC-364 cellular model nuclear transcription factor NF-κB pathway as well as TNF-α, IL-1, ICAM-1, MMP-2, MMP-3 secretion. IL-17+ TNF-α were used for stimulating RSC-364 to establish rheumatoid arthritis (RA) cellular model. Water-solubility nipponica saponin in different con-centrations was used for intervention. The influence of water-solubility nipponica saponin in different concentrations on cell viability was detected by semi-quantitative RT-PCR method. Changes in the level of TNF-α, IL-1, ICAM-1, MMP-2, and MMP-3 of culture supernatant were detected by ELISA. The results showed that the activation of NF-κB p65 in RSC-364 stimulated by TNF-α+ IL-17 can be inhibited by water-solubility nipponica saponin ac-cording to its concentration. It improved IκB-α expression, and inhibited TNF-α, IL-1, ICAM-1, MMP-2 and MMP-3 secretion. It was concluded that water-solubility nipponica saponin can inhibit the activation of NF-κB pathway, hinder the secretion and activation of multiple downstream genes, which may be its effect in inhibiting syn-ovial inflammation in RA.
RESUMO
Macrolide antibiotics are broad-spectrum, with activity against a range of Gram-positive, Gram-negative organisms and some anaerobes. The components of macrolide antibiotics are generally complicated. Therefore, it is very important to establish impurity profiles of these antibiotics to ensure their safety and process control. Compared with classical methods, the liquid chromatography-mass spectrometry method is particularly advantageous to characterize minor components at trace levels in terms of sensitivity, efficiency and selectivity, thus more and more widely used in establishments of impurity profiles. In this study, the general approaches to characterize minor components in complex pharmaceutical matrix, fragmentation pathways of 14- and 16-membered macrolide antibiotics and the establishment of the impurity profile of acetylspiramycin were given to provide valuable enlightenments to establish the impurity profiles of pharmaceutical products.
