Umbelliprenin induced both anti-inflammatory and regulatory cytokines in C57/BL6 mice.

OBJECTIVES: Umbelliprenin is a prenyloxy-coumarin with pharmacologically polyvalent activity. Several studies have shown Several studies have been shown its anti-inflammatory, anti-tumor, antioxidant, and antigenotoxic activities. However, the exact mechanism of action of this compound on the immune response has not yet been shown. Here, we investigated umbelliprenin effects on the predominance of Th1 and Th2 responses in normal C57/BL6 mice. MATERIALS AND METHODS: Umbelliprenin (2.5 mg/200 µl IP) were administered to six C57/BL6 mice every other day for 8 days. Paraffin and PBS-injected mice were enrolled as solvent and control groups, respectively (n=6 mice/group). IL-10, IFN-γ, and IL-4 levels were determined in sera and also in splenocytes culture supernatants in the presence of Con A (3 µg/ml) after 72 hr. H&E staining of paraffin embedded blocks was performed for lung and liver tissues of mice. RESULTS: Umbelliprenin could significantly increase the secretion of IFN-γ and IL-4 in sera and IL-10 in splenocytes cultures. Comparison of IFN-γ/IL-4 in the sera and splenocytes culture supernatants showed lower ratios in umbelliprenin treated mice than in solvent and untreated groups. CONCLUSION: The in vivo study showed that umbelliprenin could induce anti-inflammatory responses via the predominance of Th2 cells and some regulatory responses in C57/BL6 mice.

Immune-associated proteins with potential in vivo anti-tumor activities are upregulated in lung cancer cells treated with umbelliprenin: A proteomic approach.

Umbelliprenin (Umb), a natural coumarin, has demonstrated anti-tumor activities, both in vitro and particularly in vivo, in several types of cancer, including lung cancer. The present study aimed to identify molecular targets of Umb using a high-throughput approach. Lung cancer cell lines, QU-DB (large-cell lung carcinoma) and A549 (adenocarcinoma), were treated with Umb. Differentially-expressed proteins were identified using two-dimensional electrophoresis coupled to mass spectrometry. In the QU-DB cells, differential expression of proteins, including downregulation of the tumorigenic protein heat shock protein 90 kDa and upregulation of the potential anti-tumor proteins Nipsnap1 and glycine-tRNA ligase (GRS), suggested that Umb is a strong anti-tumor compound. In the A549 cells, differential expression of proteins indicated possible contradictory effects of Umbregarding tumorigenesis, which included downregulation of the tumorigenic protein cyclophilin and the tumor suppressor MST, and upregulation of stathmin (tumorigenic) and calreticulin. Calreticulun, in addition to GRS in QU-DB cells, stimulates anti-tumor immune responses in vivo. To the best of our knowledge, the present study is the first to use a high-throughput approach to identify targets of Umb in cancer. These molecular targets suggested that Umb may exhibit stronger in vitro anti-tumor activity against the large-cell carcinoma model than the adenocarcinoma model. Furthermore, it has been reported that Umb exhibits higher cytotoxicity against QU-DB cells than A549 cells in vitro, and significant Umb anti-tumor activity against lung cancer in vivo, which is consistent with previously published literature. In each cell type, immune-associated molecules were upregulated, indicating that this naturally occurring compound exhibits marked anti-tumor activity in vivo. However, further studies that investigate the effect of Umb in different in vitro models of cancer are required.

Umbelliprenin induced production of IFN-γ and TNF-α, and reduced IL-10, IL-4, Foxp3 and TGF-ß in a mouse model of lung cancer.

Umbelliprenin is a member of the 7-prenyloxycoumarins with potential therapeutic properties such as cytotoxic effects on various cancer cells. The present study investigates the effect of umbelliprenin on predominance of Th1 and Th2 responses in Lewis lung cancer (LLC) mouse model. The cytotoxic effect of umbelliprenin was explored on LLC cells and mouse splenocytes by MTT assay. Mice into which LLC had been transplanted were treated with umbelliprenin on alternate days, at 2.5 mg/200 µl intraperitoneally. Foxp3, TNF-α and TGF-ß mRNA expressions were assessed in tumor and lung tissues of LLC mice. In addition, IL-10, IFN-γ and IL-4 levels were determined in sera and also in splenocyte culture supernatants at the presence of tumor cell lysate (10 µg/ml) and Con A (3 µg/ml) after 72 h. Results showed the cytotoxic effects of umbelliprenin on LLC cells (IC50 = 51.6 ± 5.4 µM) while no adverse effect was seen at this concentration on normal splenocytes. TNF-α mRNA expression in both lung and tumor tissues was increased. However, Foxp3 and TGF-ß expressions were decreased in tumor tissues. Serum level of IFN-γ was elevated in the umbelliprenin treated cancerous mice compared to the control group while IL-10 and IL-4 secretions were reduced. Tumor size was also decreased in umbelliprenin treated group. In summary, umbelliprenin has shown a partially Th1 bias with a reduction of regulatory immune response. Although the mechanism behind this action is not known, it is speculated that upon changing the Th1/Th2 balance in favour of Th1, umbelliprenin induces its antitumor activity.

Umbelliprenin is cytotoxic against QU-DB large cell lung cancer cell line but anti-proliferative against A549 adenocarcinoma cells.

BACKGROUND: Umbelliprenin is a natural compound, belonging to the class of sesquiterpene coumarins. Recently, umbelliprenin has attracted the researchers' attention for its antitumor activities against skin tumors. Its effect on lung cancer is largely unknown. The aim of our study was to investigate the effects of this natural compound, which is expected to have low adverse effects, on lung cancer. METHODS: The QU-DB large cell and A549 adenocarcinoma lung cancer cell lines were treated with umbelliprenin. IC50 values were estimated using methyl thiazolely diphenyl-tetrazolium bromide (MTT) assay, in which a decrease in MTT reduction can occur as a result of cell death or cell proliferation inhibition. To quantify the rate of cell death at IC50 values, flow cytometry using Annexin V-FITC (for apoptotic cells), and propidium iodide (for necrotic cells) dyes were employed. RESULTS: Data from three independent MTT experiments in triplicate revealed that IC50 values for QU-DB and A549 were 47 ± 5.3 µM and 52 ± 1.97 µM, respectively. Annexin V/PI staining demonstrated that umbelliprenin treatment at IC50 induced 50% cell death in QU-DB cells, but produced no significant death in A549 cells until increasing the umbelliprenin concentration to IC80. The pattern of cell death was predominantly apoptosis in both cell lines. When peripheral blood mononuclear cells were treated with 50 µM and less concentrations of umbelliprenin, no suppressive effect was observed. CONCLUSIONS: We found cytotoxic/anti-proliferative effects of umbelliprenin against two different types of lung cancer cell lines.

CTLA4 gene variations and haplotypes in patients with lung cancer.

The CTLA4 protein downmodulates and terminates immune responses by sending inhibitory signals to activated T cells. In this study, six main single-nucleotide polymorphisms of the CTLA4 gene were investigated in 127 lung cancer patients and 124 healthy control subjects: -1722T/C, -1661 A/G, -318 C/T, +49A/G, +1822 C/T, and +6230 A/G (CT60). Statistical analyses revealed no significant differences in the frequencies of genotypes, alleles, and haplotypes between patients and control subjects. We also could not find significant association between CTLA4 variants and any defined lung tumor type. These six single-nucleotide polymorphisms in CTLA4 were not associated with susceptibility to lung cancer in Iranian population.

PDCD1, CTLA-4 and p53 gene polymorphism and susceptibility to gestational trophoblastic diseases.

OBJECTIVE: Gestational trophoblastic neoplasms (also termed gestational trophoblastic diseases [GTDs]) encompass a spectrum of interrelated tumors originating from trophoblasts. The search is ongoing for identification of the culpable gene defects in GTDs. Considering the role of PDCD1, CTLA-4 and p53 genes in immune regulation and tumor progression, we explored the association of single-nucleotide polymorphisms (SNPs) corresponding to each gene and GTDs. STUDY DESIGN: In a genetic association study, PD1.5 (7785) C/T, CTLA-4 +49 A/G, and p53 codon 72 Arg/Pro SNPs were genotyped in case-control groups with patient/control ratios of 92:295, 83:84 and 85:150, respectively. RESULTS: The C/T genotype of the PDCD1 gene was significantly more prevalent among patients with GTDs (40.2%) than controls (19%) (odds ratio [OR] = 2.87; 95% CI = 1.72, 4.77; p < 0.001). Moreover, the C allele was present in 65.8% of patients and 49.5% of controls (OR = 1.96; 95% CI = 1.38, 2.76; p < 0.001). There was no difference in the distribution of each genotype or allele between patients with GTDs and controls considering other studied SNPs. CONCLUSION: The results of the current study demonstrate that SNPs in the PDCD1 gene confer susceptibility to GTDs, while there is no association between CTLA-4 and p53 gene polymorphisms and GTDs in an Iranian population.