“Antivirals” in the Treatment of Adult T Cell Leukaemia– Lymphoma (ATLL)

Adult T cell leukaemia / lymphoma (ATLL) is a mature (post thymic) T cell lymphoma caused by the human T-lymphotropic virus type 1 (HTLV-1) infection. Overall survival in the aggressive subtypes (Acute Leukaemia and Lymphomatous) remains poor in part due to chemotherapy resistance. To improve treatment outcome for de novo disease, better induction therapies are required and since the pathogenic agent is known it would seem sensible to target the virus. In a recent meta-analysis the use of zidovudine and interferon alpha (ZDV/IFN) has been associated with improved response rates and prolonged overall survival in leukemic subtypes of ATLL (both acute and Chronic) confirmed in a multivariate analysis. In a more recent UK study the overall response rate for patients with aggressive ATLL treated with chemotherapy alone was 49?% compared to 81?% with combined first line therapy (chemotherapy with concurrent or sequential ZDV/IFN). Combined first line therapy prolonged median OS in acute (p?=?0.0081) and lymphomatous ATLL (p?=?0.001).These data support the use of low dose ZDV/IFN with chemotherapy as first line treatment for patients with newly diagnosed aggressive ATLL. Although the mechanisms of action are incompletely understood, some possible explanations for their efficacy will be discussed.     

Effect of 5-aza-2’-deoxycytidine on Cell Proliferation of Nonsmall Cell Lung Cancer Cell Line A549 Cells and Expression of the TFPI-2 Gene

Objective: The present study employed 5-aza-2’-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer(NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. Methods:Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 μmol/L 5-Aza-CdR, a specificdemethylating agent, for 24 ,48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM)to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), realtime polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 genemethylation status, mRNA expression and protein expression. Results: MTT assay showed that the growth ofA549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group(0 μmol/L 5-Aza-CdR). After treatment with 0, 1, 5, 10 μmol/L 5-Aza-CdR for 72h, FCM showed their proportionin G0/G1 was 69.7±0.99%, 76.1±0.83%, 83.8±0.35%, 95.5±0.55% respectively (P<0.05), and the proportion in Swas 29.8±0.43%, 23.7±0.96%, 15.7±0.75%, 1.73±0.45%, respectively (P<0.05), suggesting 5-Aza-CdR treatmentinduced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 genewas detected in control group (0 μmol/L 5-Aza-CdR), and demethylation appeared after treatment with 1, 5,10 μmol/L 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were1±0, 1.49±0.14, 1.86±0.09 and 5.80±0.15 (P<0.05) respectively. Western blotting analysis showed the relativeexpression levels of TFPI-2 protein were 0.12±0.01, 0.23±0.02, 0.31±0.02, 0.62±0.03 (P<0.05). TFPI-2 proteinexpression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration.Conclusions: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expressionin the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation ofTFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinicaltreatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be onemolecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.     

Apoptotic DNA Endonuclease (DNase-γ) Gene Transfer Induces Cell Death Accompanying DNA Fragmentation in Human Glioma Cells

Aims: Both the genetic restoration of the apoptotic pathway and the introduction of proapoptotic molecules are now drawing attention. Concerning apoptosis of human glioma cells induced by human interferon- protein, we found that DNA endonuclease (DNase-) acts as an executive molecule. The authors investigated whether gene transfer of this DNase- exerts some therapeutic effects on human glioma cells. Methods: We transduced U251SP, U251MG, and T98G human glioma cells with DNase- gene via multilamellar cationic liposomes, monitored the growth of those cells, and carefully observed the cell-death pattern. Results: DNase- gene transfer resulted in an overexpression of DNase- protein and induced DNA fragmentation in gene-transferred cells. The cytotoxic effect rose with multiple inoculations of the liposome, suggesting a relationship between its expression and the therapeutic effect. Conclusions: These results demonstrate that DNase- gene transfer can induce apoptosis in human glioma cells, indicating its potential to become a future gene therapy strategy.     

Inhibitory Effects of β-Cyclodextrin-Helenalin Complexes on H-TERT Gene Expression in the T47D Breast Cancer Cell Line - Results of Real Time Quantitative PCR

Background: Nowadays, the encapsulation of cytotoxic chemotherapeutic agents is attracting interest as amethod for drug delivery. We hypothesized that the efficiency of helenalin might be maximized by encapsulationin β-cyclodextrin nanoparticles. Helenalin, with a hydrophobic structure obtained from flowers of Arnicachamissonis and Arnica Montana, has anti-cancer and anti-inflammatory activity but low water solubility andbioavailability. β-Cyclodextrin (β-CD) is a cyclic oligosaccharide comprising seven D-glucopyranoside units,linked through 1,4-glycosidic bonds. Materials and Methods: To test our hypothesis, we prepared β-cyclodextrinhelenalincomplexes to determine their inhibitory effects on telomerase gene expression by real-time polymerasechain reaction (q-PCR) and cytotoxic effects by colorimetric cell viability (MTT) assay. Results: MTT assayshowed that not only β-cyclodextrin has no cytotoxic effect on its own but also it demonstrated that β-cyclodextrinhelenalincomplexes inhibited the growth of the T47D breast cancer cell line in a time and dose-dependent manner.Our q-PCR results showed that the expression of telomerase gene was effectively reduced as the concentrationof β-cyclodextrin-helenalin complexes increased. Conclusions: β-Cyclodextrin-helenalin complexes exertedcytotoxic effects on T47D cells through down-regulation of telomerase expression and by enhancing Helenalinuptake by cells. Therefore, β-cyclodextrin could be superior carrier for this kind of hydrophobic agent.     

Increased Risk of Developing Hepatocellular Carcinoma Associated with Carriage of the TNF2 Allele of the −308 Tumor Necrosis Factor-α Promoter Gene

BACKGROUND AND AIMS: Tumor necrosis factor-alpha (TNF-alpha) is a cytokine that may act as an endogenous tumor promoter. A genetic polymorphism of TNF-alpha at position -308 of the promoter region, which includes TNF1 (-308G) and TNF2 (-308A) alleles, has been found to be associated with susceptibility to various types of cancer. We conducted a study to evaluate the association between this polymorphism and hepatocellular carcinoma (HCC). METHODS: We recruited 74 HCC patients and 289 healthy controls, and determined their -308 TNF-alpha promoter genotypes through polymerase chain reaction followed by electrophoresis. RESULTS: Carriage of the TNF2 allele was associated with an increased risk of HCC (odds ratio [OR] = 3.5; 95% confidence interval [CI]:[2.1, 6.0]), and a trend toward a significant increase in the risk of developing HCC was observed from TNF1/TNF1, TNF1/TNF2, to TNF2/TNF2 genotypes (p < 0.01). After adjustment for gender, age, and markers of hepatitis B and C, the OR of developing HCC associated with TNF2 allele carriage was 5.3 (95% CI: [2.3, 12.1]; p < 0.01) CONCLUSIONS: Carriage of the TNF2 allele is a significant predictor of HCC independent of hepatitis B and C, and therefore it may be used as a biomarker for susceptibility to HCC.     

The Effect of 17 β-Estradiol on Invasion by the Ovarian Clear Cell Adenocarcinoma Cell Line ES-2 and the Molecular Mechanism Involved

OBJECTIVE To assess the effect of 17 β-estradiol（E2） on cell proliferation, cell invasiveness and its regulation of MTA3, Snail and matrix metalloproteinase 2 （MMP-2） expression in the ovarian clear cell adenocarcinoma cell line ES-2, and to further investigate the mechanism involved. METHODS We first investigated expression of ERα, ERβ, PR and E-cadherin of ES-2 cells by RT-PCR and Western blots. Before all experiments, the ES-2 cells were grown in medium depleted of steroid for more than 7 days. Following treatment with 10^-7,10^-8 and 10^-9 M E2, cell viability of the ES-2 cells was determined by the MTT method, and the cell cycle distribution and apoptosis were examined by flow cytometry （FCM）. Invasion and mobility assays were performed using modified Boyden chambers. MTA3, Snail and MMP-2 mRNA expression was measured by RT-PCR, and Snail, MMP-2 protein levels were determined by IHC. MMP-2 activity was assayed by zymography. RESULTS RT-PCR and Western Blots showed that theexpression of ERα and E-cadherin mRNA and protein in the ES-2 cells was negative, while ERβ and PR expression was positive. E2 at 10^-7,10^-8 or 10^-9M stimulated cell proliferation. A level of 10^-8M E2 reduced the proportion of G0-G1 phase cells and increased the proportion of cells in the S phase, but it had no effect on apoptosis. Invasiveness and mobility of the ES-2 cells was significantly increased by 10^-8M E2. Treatment with 10^-8M E2 led to reduced MTA3 mRNA expression, and elevated Snail and MMP-2 mRNA and protein levels. CONCLUSION E2 enhanced invasion by the ES-2 cells. The effects observed maybe mediated by down-regulation of MTA3 and up-reguation of Snail and MMP-2.     

The Effect of Combining Fast Neutron and Photon Irradiation on the Human Osteosarcoma OS-732 Cell Line

OBJECTIVE To determine the lethal effect of combining fast neutron with photon radiation on the OS-732 cell line. METHODS We examined the effect of irradiation by fast neutrons, photons and a mixed beam (fast neutrons plus photons) on the lethality and colony forming ability of the OS-732 cell line at different times. RESULTS Following a single irradiation close, the lethality was markedly strong at 24, 48 and 72 h in the group treated with fast neutrons alone and in the mixed beam group in which there was a high proportion of fast neutrons. CONCLUSION The lethal effect of a fast neutron and mixed beam with a high proportion of fast neutrons on the OS-732 cell line is highly significant. These studies provide guidance for the clinical application of fast neutrons for osteosarcoma treatment.     

Crocin from Kashmiri Saffron (Crocus sativus) Induces in Vitro and in Vivo Xenograft Growth Inhibition of Dalton ’ s Lymphoma (DLA) in Mice

In this study we investigated in vitro and in vivo xenograft growth inhibition by crocin isolated from Kashmirisaffron (Crocus sativus). It was found that crocin decreased cell viability in DLA cells, in a concentration- andtime-dependent manner. Significant increase in the lifespan of Dalton’s lymphoma bearing animals was notedby 37% and 44%, respectively. Furthermore, animals given treatment before induction of cancer showed 58%increase in lifespan and there was 95.6% reduction of solid tumor in crocin treated animals on the 31st day aftertumor inoculation. Crocin also showed significant impact on hematological parameters, like the hemoglobincount and numbers of lymphocytes. These findings support the conclusion that crocin from Crocus sativus hassignificant anti-tumor activity.     

Diversity of Structures Carrying the aac(6′)-aph(2“) Gene in Clinical Enterococcus faecalis and Enterococcus faecium Strains Isolated in Tunisia

Abstract

The diversity of structures carrying the aac(6′)-aph(2”) gene was studied in 46 high-level gentamicin-resistant Enterococcus faecalis and Enterococcus faecium clinical strains recovered in a Tunisian hospital during the period 2000-2003. The inclusion of the aac(6′)-aph(2”) gene within the Tn4001 composite element or in its truncated forms (lacking the IS256 at the right, the left or at both sides of the aac(6′)-aph(2”) gene) was investigated by PCR and sequencing. The aac(6′)-aph(2”) gene was included in the composite Tn4001 element in 19 of 34 high-level gentamicin-resistant E. faecalis strains (56%) and in 1 of 12 E. faecium strains (12%). A truncated form of Tn4001 lacking IS256 at the left-hand (in 10 E. faecalis and 8 E. faecium), at the right-hand (3 E. faecalis and 2 E. faecium) or at both sides of the aac(6′)-aph(2”) gene (in 2 E. faecalis and 1 E. faecium) was also detected in 26 of our enterococci. The transference by conjugation of the aac(6′)- aph(2”) gene, associated with other resistance genes, was demonstrated in seven of the high-level gentamicin-resistant E. faecalis strains.     

Paraneoplastic Syndromes of Hypercalcemia and Leukocytosis Associated with Colonic Metastases from Squamous Cell Carcinoma of the Lung： a Case Report

Lung cancer is the most common cancer-related death in both men and women in the world. Approximately 25% of all cancer deaths are attributable to lung carcinoma. Moreover, about one-half of patients with lung cancer have metastases at the time of initial diagnosis, most frequently of lymph nodes, adrenals, liver, bone and brain.     

TGF-² Signaling and the Role of Inhibitory Smads in Non-small Cell Lung Cancer

The signaling pathway mediated by transforming growth factor-² (TGF-²) participates in various biologic processes, including cell growth, differentiation, angiogenesis, apoptosis, and extracellular matrix remodeling. In the context of cancer, TGF-² signaling can inhibit tumor growth in early-stage tumors. However, in late-stage tumors, the very same pathway promotes tumor invasiveness and metastasis. This paradoxical effect is mediated through similar to mothers against decapentaplegic or Smad protein dependent and independent mechanisms and provides an opportunity for targeted cancer therapy. This review summarizes the molecular process of TGF-² signaling and the changes in inhibitory Smads that contribute to lung cancer progression. We also present current approaches for rational therapies that target the TGF-² signaling pathway in cancer.     

Associations of the -238(G/A) and -308(G/A) TNF-α Promoter Polymorphisms and TNF-α Serum Levels with the Susceptibility to Gastric Precancerous Lesions and Gastric Cancer Related to Helicobacter pylori Infection in a Moroccan Population

Objective: Helicobacter pylori (H. pylori) induces the production of tumor necrosis factor-alpha (TNF-α), which is closely related to a gastric epithelial injury. TNF-α gene polymorphism and TNF-α serum levels are associated with various malignant conditions. Identification of the ideal marker for gastric cancer (GC) is still the leading aim of several trials. Physio-pathological considerations of GC led us to investigate the association of two TNF-α promoter polymorphisms (-308G>A and -238G>A), and TNF-α serum levels with the susceptibility to gastric precancerous (PL) and GC. Methods: Patients suffering from gastric lesions (65 chronic gastritis, 50 PL, 40 GC) related to H. pylori ‎infection , and 63 healthy controls (HC) were involved in this study. Individuals are genotyped by TNF-α gene promoter sequencing and TNF-α serum levels are measured by ELISA quantitative method. Results: Regarding TNF-α-308 G/A locus, we noticed higher risk for GC (OR=4.3, CI 1.5-11.9, p-value=0.005)  and PL (OR=3.4, CI 1.2-9.2, p-value=0.01) for individuals with AA/GA genotypes compared to GG genotype. Concerning TNF-α-238 G/A locus, we noticed higher  risk for GC (OR=5.9, CI 1.2-27.5, p-value=0.01) and PL (OR=4.8, CI 1.3-18, p-value=0.01) for individuals with GG genotype compared to AA/GA genotypes. We noticed that TNF-α serum levels have been increased together with gastric lesions severity. Moreover, TNF-α-308 and TNF-α-238 A alleles seemed to, respectively, upregulate and downregulate TNF-α serum levels. Conclusion: The TNF-α -308 A allele has a promotive effect for GC progression, whereas the TNF-α -238 A allele has a protective function against GC progression. High levels of TNF-α seemed to be associated with the aggressiveness of gastric lesions. TNF-α gene polymorphisms and TNF-α serum levels might be helpful to select those patients who are at high risk for GC.