RESUMO
BACKGROUND: Lung cancer is a common and deadly disease. Chemotherapy is the most common treatment, which inhibits cancer cell growth. Pemetrexed (PMX) is often used with other drugs. Environmental stress can affect regulatory non-coding RNAs such as MicroRNAs that modify gene expression. This study investigates the effect of PMX on the hsa-miR-320a-3p expression in the Calu-6 lung cancer cell line. METHODS AND RESULT: Calu-6 cells were cultured in an incubator with 37 °C, 5% CO2, and 98% humidity. The MTT test was performed to determine the concentration of PMX required to inhibit 50% of cell growth. To examine growth inhibition and apoptosis, release of lactate dehydrogenase (LDH), cell assays and caspase 3 and 7 enzyme activity were used. Finally, molecular studies were conducted to compare the expression of hsa-miR-320a-3p and genes including VDAC1, DHFR, STAT3, BAX and BCL2 before and after therapy. RESULTS: According to a study, it has been observed that PMX therapy significantly increases LDH release after 24 h. The study found that PMX's IC50 on Calu-6 is 8.870 µM. In addition, the treated sample showed higher expression of hsa-miR-320a-3p and BAX, while the expression of VDAC1, STAT3, DHFR and BCL2 decreased compared to the control sample. CONCLUSION: According to the findings of the current research, hsa-miR-320a-3p seems to have the potential to play an important role in the development of novel approaches to the treatment of lung cancer.
Assuntos
Neoplasias Pulmonares , MicroRNAs , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Pemetrexede/farmacologia , Regulação para Cima/genética , Proteína X Associada a bcl-2/genética , MicroRNAs/genética , Linhagem CelularRESUMO
Rhamnolipids produced by P. aeruginosa MR01 were fractionated into mono- and di-rhamnolipids, and their dominant congeners, Rha-C10-C10 and Rha-Rha-C10-C10, were shown by mass spectrometry. Minimum surface tensions and critical micelle concentrations (CMC) were determined as "≃34â¯mN/m; ≃26.17â¯mg/l;" and "≃29â¯mN/m; ≃29.63â¯mg/l" for mono- and di-rhamnolipids, respectively. Spectrophotometry measurements provided a close approximation of CMC. Contact angle and diameter of wet area were determined for rhamnolipid-containing drops on hydrophobic paper to display their capability for alteration of surface wettability. Wet area measurement is a simple, reliable method not requiring a Drop Shape Analyzer. Cell viabilities determined by MTT assay showed a decline in a dose-dependent manner and estimated IC50 values were 25.87⯵g/ml and 31.00⯵g/ml for mono- and di-rhamnolipids treating MCF-7 cells for 48â¯h. Morphological observations using the inverted phase-contrast microscopy and fluorescence microscopy via Hoechst staining revealed the apoptotic characteristics in treated MCF-7 cells. The semi-quantitative RT-PCR method demonstrated that expression of the p53â¯gene in mRNA levels significantly (Pâ¯<â¯0.05) increased when treated with 30⯵g/ml of each rhamnolipid compound for 12â¯h. It can be concluded that rhamnolipids derived from MR01 show significant anticancer potential against MCF-7 cell line and should be further investigated as natural, therapeutic anti-tumor agents.