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PURPOSE: The long non-coding RNA H19, a conservatively imprinted gene, acts as a molecular sponge for the let-7 family, which has been identified as a set of tumor suppressors. However, the combined prognostic value of H19 and let-7a signature in breast cancer patients remains unclear. METHODS: In this research we assessed the prognostic value of the combined H19 and let-7a signature in breast cancer patients by retrospectively reviewing that data of 79 patients who underwent neoadjuvant chemotherapy; we also investigated the expression and function of H19 in breast cancer cell lines in vitro. Survival data were calculated using the Kaplan-Meier method and compared using the log-rank test. Univariate and multivariate survival analyses were conducted using the Cox proportional hazards regression method. As determined using X-tile, the optimal cutoff value for the risk score to assess progression-free survival (PFS) based on the combined signature was –0.1. RESULTS: Patients with an overall positive treatment response had higher let-7a and lower H19 levels. In addition, let-7a expression was negatively correlated with H19 expression. Patients with a risk score of >–0.1 had shorter overall survival and PFS. In vitro data showed that chemoresistant cell lines exhibit higher H19 and lower let-7a levels and knockdown H19 restores paclitaxel sensitivity. CONCLUSION: Our results suggest that the combined let-7a and H19 signature is a novel prognostic factor for breast cancer patients treated with neoadjuvant chemotherapy.
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Humans , Breast Neoplasms , Breast , Cell Line , Disease-Free Survival , Drug Therapy , In Vitro Techniques , Methods , Neoadjuvant Therapy , Paclitaxel , Porifera , Prognosis , Retrospective Studies , RNA, Long NoncodingABSTRACT
This study was aimed to examine the effect of Shaoyao Tang (SYT) on the treatment of dextran sulfate sodium (DSS)-induced experimental colitis in mice via targeting the Notch signaling pathway.The mice were equally and randomly divided into the normal control group (control),model group (DSS),DSS + SYT group (17.8 g· kg-1),n=10.Mice were administered with 3.5％ DSS for 7 days for the model establishment.And then,SYT was given to the model group on the second day after the induction of colitis with DSS for 7 days.The effects were studied by the disease activity index (DAI),histological analysis by HE staining,myeloperoxidase (MPO) activity.ELISA was used to measure secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the colon tissues.The expression of mRNA and protein of Notch-1,Hes-1,Math-1 and Mucin-2 (Muc-2) were detected using real-time PCR and immunohistochemistry.The results showed that compared with the model group,SYT attenuated symptoms of colitis by decreasing DAI score,histopathologic score and MPO activity,as well as increasing the colon length.SYT treatment also inhibited the production of IL-6 and TNF-a in colon tissues of mice.Furthermore,SYT significantly blocked the expression of mRNA and protein of Notch-1 and Hes-1,but increased the expression of Math-1 and Muc-2.It was concluded that SYT exerted the anti-inflammatory activity in mice model of DSS-induced colitis by inhibiting the Notch signaling pathway.
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Background:Tryptophan(TRP)is an essential amino acid,and can produce 5-hydroxytryptamine(5-HT)via 5-HT signal pathway and kynurenine( KYN)metabolic pathway under the catalysis of enzyme,thereby participating in the pathogenesis of visceral hypersensitivity in diarrhea-predominant irritable bowel syndrome(IBS-D). Aims:To investigate the relationship between visceral sensitivity and TRP metabolic pathway in patients with IBS-D. Methods:Thirty patients with IBS-D and 30 healthy controls from June 2012 to January 2014 at Guangdong Provincial TCM Hospital were enrolled. Score of gastrointestinal symptom rating scale( GSRS)was evaluated. Visceral sensitivity was measured by anorectal manometry. RT-PCR and Western blotting were used to detect mRNA and protein expressions of colon mucosal IDo, respectively. Serum 5-HT,5-HIAA,TRP,IDo,KYN,KYNA concentrations and IDo activity,KAT activity were determined by high performance liquid chromatography assay. Results:Compared with control group,GSRS score was significantly increased(P < 0. 05),initial perception threshold,defecation threshold,pain threshold were significantly decreased(P < 0. 05),anorectal constriction pressure was significantly increased( P < 0. 05),serum 5-HT,5-HIAA concentrations were significantly increased(P < 0. 05),mRNA and protein expressions of IDo were significantly increased (P < 0. 05),serum KYN was significantly increased(P < 0. 05),KYNA was significantly decreased(P < 0. 01),IDo activity was significantly incseased(P < 0. 01),and KAT activity was significantly decreased in IBS-D group(P < 0. 01). Correlation analysis showed that initial perception threshold,defecation threshold,pain threshold and anorectal constriction pressure were correlated with 5-HT,5-HIAA,TRP,KYN,KYNA,IDo activity and KAT activity in patients with IBS-D (P < 0. 05). Conclusions:TRP metabolic pathway is correlated with visceral hypersensitivity in patients with IBS-D.
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Objective To investigate the cytokines levels in peripheral blood of ulcerative colitis (UC) patients at active stage, and to observe the effect of baicalin at various concentrations on the cytokines. Methods Quantitative polymerase chain reaction (Q-PCR) assay was used for the detection of interleukin 4 receptor (IL4R), IL6R, IL23R and RAR-related orphan receptor C (RORC) expression in single peripheral mononuclear cell of UC patients. Enzyme-linked immunosorbent assay (ELISA) was used to examine the serum levels of interferon gamma (IFN-γ), IL-4, IL-5, IL-6, IL-10, and transforming growth factor beta (TGF-β) levels in UC patients, and was applied for in-vitro detection of these indicators after baicalin intervention. Results Q-PCR results showed that IL4R, IL6R, IL23R, RORC gene expression levels in UC patients were increased to various degrees as compared to diarrhea-predominant irritable bowel syndrome (IBS-D) patients and the healthy volunteers, and then the levels were decreased to various degrees after intervention by baicalin at different concentrations, in particular at 20 and 40μmol/L. The results of ELISA showed that serum levels of cytokines of IFN-γ, IL-5, IL-6 in UC patients were increased to various degrees while IL-4, IL-10 and TGF-β1 was decreased as compared to IBS-D patients and the healthy volunteers. Baicalin at the concentrations of 20 and 40 μmol/L had an effect on decreasing IFN-γ, IL-5, IL-6 and on increasing IL-4 and IL-10. Conclusion Higher concentrations of baicalin show obvious effect on inhibiting RORC and IL23R expression, on decreasing IFN-γ, IL-5, IL-6 levels, and on increasing IL-4, IL-10 and TGF-β1 levels, which indicated that the therapeutic mechanism of baicalin in relieving ulcerative colonic inflammation is related with the regulation of immune function.
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Metastasis is a multistep process involving modification of morphology to suit migration, reduction of tumor cell adhesion to the extracellular matrix, increase of cell mobility, tumor cell resistance to anoikis, and other steps. MicroRNAs are well-suited to regulate tumor metastasis due to their capacity to repress numerous target genes in a coordinated manner, thereby enabling their intervention at multiple steps of the invasion-metastasis cascade. In this study, we identified a microRNA exemplifying these attributes, miR-124, whose expression was reduced in aggressive MDA-MB-231 and SK-3rd breast cancer cells. Down-regulation of miR-124 expression in highly aggressive breast cancer cells contributed in part to DNA hypermethylation around the promoters of the three genes encoding miR-124. Ectopic expression of miR-124 in MDA-MB-231 cells suppressed metastasis-related traits including formation of spindle-like morphology, migratory capacity, adhesion to fibronectin, and anoikis. These findings indicate that miR-124 suppresses multiple steps of metastasis by diverse mechanisms in breast cancer cells and suggest a potential application of miR-124 in breast cancer treatment.
Subject(s)
Female , Humans , Anoikis , Breast Neoplasms , Genetics , Metabolism , Pathology , Cell Adhesion , Cell Line, Tumor , Cell Movement , Connective Tissue Growth Factor , Metabolism , DNA Methylation , Down-Regulation , Gene Expression Regulation, Neoplastic , MicroRNAs , Genetics , Metabolism , Neoplasm Metastasis , rho GTP-Binding Proteins , Metabolism , rho-Associated Kinases , MetabolismABSTRACT
AIM: To detect the effect of conjunction matrigel with mammary fad pat(MFP)implantation on the tumorigenesis, proliferation, apoptosis and metastasis of Her2 positive and negative breast cancer model. METHODS: The Her2 positive BT 474 and Her2 negative MDA-MB 231 breast cancer cells were injected into MFP of nude mice with or without matrigel to establish breast cancer model. The tumor volume was measured every 3 d. Followed up for 30 d after implantation, the nude mice were killed and the tumors and associated organs were dissected for pathological sectioning and staining with hematoxylin and eosin. The time of tumor formation and the tumorigenesis were determined after implantation. The tumor volume and metastasis rate were calculated and compared with each other. The proliferation and apoptosis of Her2 positive and negative tumors were also determined. RESULTS: Matrigel and MFP implantation technology shortened the time of tumorigenesis significantly(P<0.01). The tumorigenesis rate of BT 474 and MDA-MB 231 breast cancer cells did not show any different(P>0.05). The metastasis rate of MDA-MB 231 breast cancer cells were improved from 25.0% to 37.5%(P<0.05). CONCLUSION: Matrigel and MFP implantation can be combined to shorten the time of tumor formation by two kinds of breast cancer cells, and improve the metastasis of Her2 negative MDA-MB 231 cells. Using matrigel does not show any effect of proliferation and apoptosis on Her2 positive and negative breast cancer cells.