ABSTRACT
OBJECTIVES: Cancer stem cells (CSCs) comprise a small population of cells in cancerous tumors and play a critical role in tumor resistance to chemotherapy. miRNAs have been reported to enhance the sensitivity of pancreatic cancer to chemotherapy. However, the underlying molecular mechanism requires better understanding. METHODS: Cell viability and proliferation were examined with CCK8 assays. Quantitative real-time polymerase chain reaction was executed to assess mRNA expression. StarBase database was used to select the target genes of miRNA, which were further affirmed by dual luciferase assay. Transwell assay was used to analyze cell invasion and migration. RESULTS: We proved that miR-497 could be obviously downregulated in pancreatic cancer tissues and CSCs from Aspc-1 and Bxpc-3 cells. In addition, inhibition of miR-497 evidently accelerated pancreatic CSC gemcitabine resistance, migration and invasion. Moreover, we revealed that nuclear factor kappa B 1 (NFκB1) was prominently upregulated in pancreatic cancer tissues and pancreatic CSCs, and NFκB1 was also identified as a direct target of miR-497. Furthermore, we demonstrated that overexpression of NFκB1 could also notably promote the viability, migration, and invasion of gemcitabine-treated pancreatic CSCs, but this effect could be partially abolished by miR-497 overexpression. CONCLUSIONS: Those findings suggest that miR-497 overexpression could suppress gemcitabine resistance and the metastasis of pancreatic CSCs and non-CSCs by directly targeting NFκB1.
Subject(s)
MicroRNAs , Pancreatic Neoplasms , Cell Line, Tumor , Cell Proliferation/genetics , Deoxycytidine/analogs & derivatives , Drug Resistance, Neoplasm/genetics , Gene Expression Regulation, Neoplastic , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , NF-kappa B/metabolism , Neoplastic Stem Cells/metabolism , Pancreatic Neoplasms/drug therapy , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/metabolism , Gemcitabine , Pancreatic NeoplasmsABSTRACT
PURPOSE: Explore the effect of Baihe Dihuang powder on chronic stress depression rat models. METHODS: Chronic stress depression rat models were established with different stimuli for 21â¯days. At the same time, the drug was administered for 21 consecutive days. The animals were weighed once a week after the start of the formal experiment. On the second day after the end of drug administration, conduct sugar water consumption test and open-filed box experiment, and conduct behavioral observation; At the end of behavioral testing, blood was taken from the eyeball and plasma was separated to measure MDA level and erythrocyte SOD activity; Take brain for homogenate, then measure the contents of 5-HT, NE and DA in brain tissue homogenate; Take the thymus and spleen, stained with 10% formalin fixation, embedding and HE staining, then use microscope to observe the histopathological changes. RESULTS: Chronic stress depression rats model replicated successfully. Each group of given drugs could increase the weight, the consumption of sugar water, and improve the behavioral score, increase erythrocytes SOD activity and decrease MDA level of plasma, increase the content of 5-HT, NE and DA of brain homogenate, and improve the pathological changes of thymus and spleen of chronic stress depression model animals. CONCLUSION: Chronic stress depression rat model replicates successfully. Baihe Dihuang powder can interfere chronic stress depression rats model through different action pathways.