anisomycin and Neoplasm-Metastasis

Either apigenin or chrysin alone has been found to exert anti-inflammatory and tumor suppressive effect. However, the combined effect of apigenin and chrysin on colorectal cancer (CRC) has not been fully clarified. We attempted to explore the effect of chrysin and apigenin on CRC and its related mechanism. SW480 and HCT-116 cells were treated with either apigenin or chrysin alone or two-drug combination at different doses of 5, 25, 50, 100 μM for optimal concentration determination. Then, we focused on the individual and combined effect of apigenin and chrysin on clonogenicity, apoptosis, metastasis-related behaviors of CRC cells by colony formation assay, cell scratch assay, flow cytometry, and transwell assay. The changes of the activation of P38-MAPK/AKT pathway were evaluated underlying apigenin and chrysin intervention, further after co-treated with P38-MAPK agonist anisomycin. Apigenin (25 μM) combined with chrysin (25 μM) were determined to be optimal. Treatment with the combination of apigenin (25 μM) and chrysin (25 μM) significantly reduced cell clone numbers, migration, and invasion ability, while increased the cell apoptosis in both CRC cell lines. The combined effect was higher than chrysin or apigenin alone. Meanwhile, p-P38 and p-AKT were significantly downregulated by chrysin and apigenin treatment. The tumor inhibitive effect of apigenin combined with chrysin was obviously reversed by adding P38 agonist, anisomycin. Apigenin (25 μM) combined with chrysin (25 μM) showed synergetic effect in inhibiting the growth and metastasis of CRC cells by suppressing the activity of P38-MAPK/AKT pathway.

Topics: Adenocarcinoma; Anisomycin; Apigenin; Apoptosis; Cell Line, Tumor; Cell Movement; Clone Cells; Colorectal Neoplasms; Drug Synergism; Flavonoids; HCT116 Cells; Humans; MAP Kinase Signaling System; Molecular Targeted Therapy; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Proteins; p38 Mitogen-Activated Protein Kinases; Proto-Oncogene Proteins c-akt; Tumor Stem Cell Assay

MALAT1 (metastasis associated lung adenocarcinoma transcript1) is a conserved long non-coding RNA, known to regulate gene expression by modulating transcription and post-transcriptional pre-mRNA processing of a large number of genes. MALAT1 expression is deregulated in various tumors, including breast cancer. However, the significance of such abnormal expression is yet to be fully understood. In this study, we demonstrate that regulation of aggressive breast cancer cell traits by MALAT1 is not predicted solely based on an elevated expression level but is context specific. By performing loss- and gain-of-function studies, both under in vitro and in vivo conditions, we demonstrate that MALAT1 facilitates cell proliferation, tumor progression and metastasis of triple-negative breast cancer (TNBC) cells despite having a comparatively lower expression level than ER or HER2-positive breast cancer cells. Furthermore, MALAT1 regulates the expression of several cancer metastasis-related genes, but displays molecular subtype specific correlations with such genes. Assessment of the prognostic significance of MALAT1 in human breast cancer (n=1992) revealed elevated MALAT1 expression was associated with decreased disease-specific survival in ER negative, lymph node negative patients of the HER2 and TNBC molecular subtypes. Multivariable analysis confirmed MALAT1 to have independent prognostic significance in the TNBC lymph node negative patient subset (HR=2.64, 95%CI 1.35- 5.16, p=0.005). We propose that the functional significance of MALAT1 as a metastasis driver and its potential use as a prognostic marker is most promising for those patients diagnosed with ER negative, lymph node negative breast cancer who might otherwise mistakenly be stratified to have low recurrence risk.

Topics: Anisomycin; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epithelial-Mesenchymal Transition; Estrogen Receptor alpha; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Lymphatic Metastasis; Multivariate Analysis; Neoplasm Metastasis; Neoplasm Recurrence, Local; Phenotype; Prognosis; Risk; RNA Splicing; RNA, Long Noncoding; Triple Negative Breast Neoplasms

The STAT proteins are the mediators in the signal transduction in between extracellular environment and nucleus. Based on its own activity STATs regulate expression of genes involved in normal and pathological cellular processes. Constitutive STAT3 activation, the results of different cytokines inductions, has been shown in many primary human cancers. STAT3, as an oncogenic protein, plays an important role in the regulation of autonomous properties of cancer cells.. In this study the effectiveness of the STAT3 gene expression activity silencing with RNA interference method was assessed. pSUPER.neo shRNA coding expression vector: shRNA-STAT3 and control vectors: shRNA-SCR, and pGFP were used. Effects of silencing of the examined gene was described as the phenotype changes of modulated HeLa (CCL-2) cancer cell line. To characterize modulated cancer cells phenotype changes two methods were applied: Wound Healing Assay and the stimulation to the apoptosis with anisomycin.. According to control cells, the silencing of the STAT3 gene expression activity reduced the mobility of modulated cells as well after 24 as after 48 hours after modulation. Also, after anisomycin stimulation the increasing in apoptotic modulated cell death was seen.. The inhibition of the activity of the STAT3 gene decreases HeLa cell migration, moreover the blocked STAT3 ability to the antyapoptotic gene expression activation leads to the increased susceptibility to apoptotic cell death.

Topics: Anisomycin; Apoptosis; Cell Line, Tumor; Cell Movement; Gene Expression Regulation, Neoplastic; Gene Silencing; HeLa Cells; Humans; Neoplasm Metastasis; Signal Transduction; STAT3 Transcription Factor

Malignant epithelial cells with metastatic potential resist apoptosis that normally occurs upon loss of anchorage from the extracellular matrix, a process termed "anoikis." Resistance to anoikis enables malignant cells to survive in an anchorage-independent manner, which leads to the formation of distant metastases. To understand the regulation of anoikis, we designed, automated, and conducted a high-throughput chemical screen for anoikis sensitizers. PPC-1 anoikis-resistant prostate cancer cells were seeded in hydrogel-coated ultralow binding plates for suspension conditions and standard tissue culture plates to promote adhesion. After seeding, cells were treated with aliquots from a library of previously characterized small molecules, and viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt, assay. From this chemical screen, we identified anisomycin that induced apoptosis in suspension conditions, but was not toxic to these cells grown under adherent conditions. Anisomycin sensitized cells to anoikis by decreasing levels of the caspase-8 inhibitor FLIP and subsequently activating the death receptor pathway of caspase activation. Although anisomycin activated c-Jun-NH(2)-kinase and p38, these kinases were not functionally important for the effect of anisomycin on anoikis and FLIP. Rather, anisomycin decreased FLIP and sensitized cells to anoikis by inhibiting its protein synthesis. Finally, we showed that anisomycin decreased distal tumor formation in a mouse model of prostate cancer metastases. Thus, a novel chemical screen identified anisomycin as an anoikis sensitizer that acts by decreasing FLIP protein synthesis. Our results suggest that FLIP is a suppressor of anoikis and inhibiting FLIP protein synthesis may be a useful antimetastatic strategy.

Topics: Animals; Anisomycin; Anoikis; CASP8 and FADD-Like Apoptosis Regulating Protein; Caspases; Cell Survival; Drug Screening Assays, Antitumor; Humans; Male; Mice; Models, Biological; Neoplasm Metastasis; Neoplastic Cells, Circulating; Prostatic Neoplasms; Protein Biosynthesis; Receptors, Death Domain; Tumor Cells, Cultured