Dual anti-metastatic and anti-proliferative activity assessment of two probiotics on Hela and HT-29 cell lines

Objective: Lactobacilli are a group of probiotics with beneficial effects on prevention of cancer. However, there is scant data in relation with the impacts of probiotics in late-stage cancer progration, especially metastasis. The present original work was aimed to evaluate the anti-metastatic and anti-proliferative activity of lactobacillus rhamnosus supernatant [LRS] and lactobacillus crispatus supernatant [LCS] on the human cervical and colon adenocarcinoma cell lines [HeLa and HT-29, respectively]

Materials and Methods: In this experimental study, the anti-proliferative activities of LRS and LCS were determined through MTT assay. MRC-5 was used as a normal cell line. Expression analysis of CASP3, MMP2, MMP9, TIMP1 and TIMP2 genes was performed by quantitative reverse transcriptase-polymerase chain reaction [qRT-PCR], following the cell synchronization

Results: Supernatants of these two lactobacilli had cytotoxic effect on HeLa, however LRS treatment was only effective on HT-29 cell line. In addition, LRS had no side-effect on normal cells. It was shown that CASP3 gene expression has been reduced after treatment with supernatants of two studied lactobacilli. According to our study, LRS and LCS are efficacious in the prevention of metastasis potency in HeLa cells with decreased expression of MMP2, MMP9 and increased expression of their inhibitors. In the case of HT-29 cells, only LRS showed this effect

Conclusion: Herein, we have demonstrated two probiotics which have anti-metastatic effects on malignant cells and they can be administrated to postpone late-stage of cancer disease. LRS and LCS are effective on HeLa cell lines while only the effect of LRS is significant on HT-29, through cytotoxic and anti-metastatic mechanisms. Further assessments are required to evaluate our results on the other cancer cell lines, in advance to use these probiotics in other extensive trial studies

Association of CD58 polymorphism with multiple sclerosis and response to interferon beta therapy in a subset of Iranian population

Multiple sclerosis [MS] is one of the leading neurodegenerative causes of physical disability world-wide. Genetic aberrations of autoimmunity pathway components have been demonstrated to significantly influence MS development. Cluster of Differentiation 58 [CD58] is pertained to a group of genes which had been assayed in several recent association studies. Given the significance of CD58 in modulation of T regulatory cells that control autoimmune responses, the present study was conducted to investigate the frequency of rs12044852 polymorphism and its effect on the outcome of interferon beta [IFN- beta] therapy in a subset of Iranian MS patients. Two hundred MS patients and equal number of healthy controls were recruited to be genotyped in an experimental case-control based study through polymerase chain reaction using specific sequence primers [PCR-SSP]. Relapsing remitting multiple sclerosis [RRMS] patients administered IFN- beta therapy were followed up with clinical visits every three months up to two years. The mean of multiple sclerosis severity score [MSSS] and expanded disability status scale [EDSS] were measured to monitor the change in severity of MS in response to IFN- beta therapy. Pearson's Chi-square and analysis of variance [ANOVA] tests were the main statistical methods used in this study. Strong association was found between the CC genotype and onset of MS [p=0.001, OR=2.22]. However, there was no association between rs12044852 and various classifications and severity of MS. Pharmacogenetics-based analysis indicated that carriers of CC genotype had the highest MSSS score compared to others, implying a negative impact of rs12044852 on response to IFN- beta t herapy. Taken together, our findings revealed the critical effect of rs12044852 polymorphism of CD58 on the progression of MS disease. This indicates that genotyping of MS patients may expedite achieving personalized medical management of MS patients

Adenosine deaminase activity in estrogen receptor positive and negative human breast cancer cell lines

The aims of this study were to assay the activity of adenosine deaminase [ADA] in estrogen receptor positive [MCF-7] and negative [MDA-MB468] breast cancer cell lines.MDA-MB468 and MCF-7 breast cancer cell lines were cultured in complete medium, striped serum with and without 0.01micro M diethylstilbestrol [DBS], complete medium in the presence and absence of 1micro M tamoxifen for 20 hr. Adenosine deaminase activity was determined using the colorimetric method described by Guisti and Galanti.It was found that the activity of enzyme in estrogen receptor positive [ER+] cell line [MCF-7] was significantly higher than that of estrogen receptor negative breast cancer cell line [MDA-MB468]. ADA activity in MCF-7 cells cultured in the presence of tamoxifen or charcoal-striped serum was significantly lower than that of control. Furthermore addition of diethylstilbestrol [DBS] to the striped serum increased the value of ADA activity to that of control.Unlike MCF-7 cells, the activity of ADA in MDA-MB468 cells remained unchanged upon treatment with tamoxifen or striped serum.These findings suggest estrogen responsiveness of ADA expression in MCF-7 cells

apoptotic effect of extracellular zinc sequestration on HT29/219 and SW742 cell lines

Zn [II] is an important regulator of caspase-3, as well as an antioxidant, microtubule stabilizer, growth cofactor, and anti-inflammatory agent. Over the past 30 years, many researchers have demonstrated the important role of Zn [II] in a variety of physiological processes, including growth and development, maintenance and priming of the immune system, and in tissue repair and regeneration. In this study, we present evidence that chelation of extracellular zinc by diethylenetriaminepentacetic acid [DTPA] in different concentrations causes cell death in carcinoma cell lines, HT29/219 and SW742. Hoechst 33258 staining revealed that cell death was mainly by apoptosis. Additionally, significant increases in the activity of caspase-3 and -9 were observed in both cell lines. Caspase-8 activation was negligible in both cell lines. The cytotoxicity/apoptotic effect of DTPA was inhibited significantly by Zn [II], Cu [II] and N-Acetyl-L-Cysteine [NAC] [P<0.05]. Therefore, DTPA, the membrane-impermeable metal ion chelator, induces apoptosis through the depletion of extracellular zinc ion

Cytotoxicity effect of cladribine on the MCF-7 human breast cancer cell line

Cladribine, an analogue of deoxyadenosine, is highly toxic for both non-dividing and proliferating cells and has shown activity in the treatment of several malignancies. Therefore, the aim of the present study is to investigate the cytotoxicity effect of cladribine [2-CdA] on the breast cancer cell line, MCF-7 [estrogen receptor positive, ER+]. MTT assay, annexin V-Fluorescein/PI and Hoechst 33258 staining were used to detect cytotoxicity and cell apoptosis. The activation of caspase-3 and -9 was assayed using caspase activation assay kits. Gel electrophoresis was performed to detect DNA fragmentation. Treatment of MCF-7 cells with different concentrations of 2-CdA resulted in a significant increase in the cell death. Annexin V-Fluorescein/PI and Hoechst 33258 staining revealed that the cell death was mainly an apoptotic type. A significant [p<0.05] increase in the activity of caspase-9 was observed but Caspase-3 activity was unchanged and DNA laddering profile was not obtained. Pre-treatment of the cells with kinase inhibitor, 5 -amino-5 -deoxyadenosine inhibited the cytotoxicity effect of cladribine. In conclusion, this study has shown that high dose of cladribine [higher than 25 micro M] has an apoptotic effect on MCF-7 cells and that its intracellular phosphorylation is necessary

Cell kinetic study of tamoxifen treated MCF7 and MDAMB 468 breast cancer cell lines

Apoptosis could be a major mechanism of antitumor effect of tamoxifen. Therefore this study is designed to characterize the kinetic behavior of tamoxifen-induced apoptosis in the estrogen receptor positive [ER+] and negative [ER-] cell lines, MCF-7 and MDA-MB-468. Frequency of cell death was examined by trypan blue and acridine orange staining. Annexin V-Fluorescein/PI was used in flow cytometry for distinguishing the dividing, apoptotic and necrotic cells and Hoechst 33258 staining was also applied to detect apoptotic changes in the nuclear morphology. The results showed that tamoxifen was able to induce apoptosis in both cell lines [