Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system

Abstract

Introduction and Aim: Cancers are a complex group of genetic illnesses that develop through multistep, mutagenic processes which can invade or spread throughout the body. Recent advances in cancer treatment involve oncolytic viruses to infect and destroy cancer cells. The Newcastle disease virus (NDV), an oncolytic virus has shown to have anti-cancer effects either directly by lysing cancer cells or indirectly by activating the immune system. The green fluorescent protein (GFP) has been widely used in studying the anti-tumor activity of oncolytic viruses. This study aimed to study the anticancer effect of a recombinant rNDV-GFP clone on NCI-H727 lung carcinoma cell line in vitro. Materials and Methods: The GFP gene was inserted to a NDV strain to create a recombinant NDV (rNDV- GFP) using reverse genetics technology. The MTT assay was used in evaluating the oncolytic effect of rNDV- GFP on the lung carcinoma NCI-H727 cells. Light and fluorescent microscopy was used to study the cytopathic effects of rNDV-GFP. Results: MTT assay showed that rNDV-GPF inhibited the NCI-H727 tumor cell death in a time-dependent manner. A significant inhibitory effect (78.3%) for rNDV-GPF on cancer cells was observed at 96h in comparison to rNDV (22.7%) and the cytotoxicity rate was directly proportional to the MOI used. Microscopic studies showed rNDV-GPF to induce cytopathic effect post 24 h of infection. Conclusion: The GFP-expressing recombinant NDV strains exhibited encouraging results in terms of tumor growth inhibition. Our research set the groundwork for employing recombinant NDV as an anticancer viral vector.

Prevalence of Methicillin-Resistant and Methicillin Sensitive Staphylococcus aureus Nasal Carriage and their Antibiotic Resistant Patterns in Kirkuk City, Iraq

“…47 There are many recommendations for applying the PCR techniques in the diagnosis of Methicillin-resistant Staphylococcus aureus 48,49 due to their accurate results and short time. The polymerase chain reaction (PCR) was applied in the detection of pathogenic microorganisms, [50][51][52][53][54][55][56][57][58][59][60] and genetic-related diseases. [61][62][63][64]…”

Study of the association of toll like-4 receptor gene polymorphism (rs1927911) and susceptibility to urinary tract infection among Iraqi patients

“…DNA extracted was checked for its purity by measuring its absorbance at 260/280nm using a nanodrop. DNA extracted was subjected to amplification of SNP rs1927911 of the TLR4 gene using PCR, this technique used in the medicine area (17)(18)(19). The primers (Macrogen, Korea) given in Table 1.…”

Association between genetic variants in ANGPTL4 (T266M) and MC4R genes predisposing to obesity

“…Appearance of 163bp + 44 bp fragments referred to the presence of MC4R c.124 G > T mutation. Numerous hereditary illnesses, including adenocarcinoma (12,13), chronic myeloid leukemia (14), and beta-thalassemia (15), have been successfully diagnosed using the molecular approach and in the treatment of certain malignancies (16). On the other hand, this method was utilized to diagnose several harmful microorganisms in addition to hereditary illnesses (17), therefore, polymerase chain reaction (PCR) is recommended for detecting genetic variations in the ANGPTL4 (T266M) and MC4R genes that are associated with the diagnosis of obesity.…”