Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Khosravi, Saeed Zaka; Moonesi, Mohammadreza; Moradabadi, Alireza; Rajaeinejad, Mohsen; Heidari, Mohammad Foad; Noroozi-Aghideh, Ali

doi:10.31557/apjcp.2022.23.1.125

Cited by 6 publications

(1 citation statement)

References 35 publications

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“…Conventional chemotherapeutic agents such as cisplatin, paclitaxel, and methotrexate are cytotoxic and can directly destroy malignant cells by damaging DNA 4,5 , blocking intracellular protein synthesis 6,7 , and affecting nucleic acid metabolism thereof [8][9][10] , which are limited by the collateral damage to normal cells. Mutations in various pro-oncogenes (such as RAS, MYC, and SIS family genes) in lung cancer cells are among the important endogenous factors leading to its development [11][12][13] . Therefore, the exploration of novel anti-cancer agents with molecular targeting functions, which can also selectively inhibit lung cancer cells (low toxicity and high efficiency), is a significant entry point for the treatment of lung cancer at present.…”

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confidence: 99%

Dihydroartemisinin-driven selective anti-lung cancer proliferation by binding to EGFR and inhibition of NRAS signaling pathway-induced DNA damage

Li,

Peng,

Yang

et al. 2024

Sci Rep

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Chemotherapeutic agents can inhibit the proliferation of malignant cells due to their cytotoxicity, which is limited by collateral damage. Dihydroartemisinin (DHA), has a selective anti-cancer effect, whose target and mechanism remain uncovered. The present work aims to examine the selective inhibitory effect of DHA as well as the mechanisms involved. The findings revealed that the Lewis cell line (LLC) and A549 cell line (A549) had an extremely rapid proliferation rate compared with the 16HBE cell line (16HBE). LLC and A549 showed an increased expression of NRAS compared with 16HBE. Interestingly, DHA was found to inhibit the proliferation and facilitate the apoptosis of LLC and A549 with significant anti-cancer efficacy and down-regulation of NRAS. Results from molecular docking and cellular thermal shift assay revealed that DHA could bind to epidermal growth factor receptor (EGFR) molecules, attenuating the EGF binding and thus driving the suppressive effect. LLC and A549 also exhibited obvious DNA damage in response to DHA. Further results demonstrated that over-expression of NRAS abated DHA-induced blockage of NRAS. Moreover, not only the DNA damage was impaired, but the proliferation of lung cancer cells was also revitalized while NRAS was over-expression. Taken together, DHA could induce selective anti-lung cancer efficacy through binding to EGFR and thereby abolishing the NRAS signaling pathway, thus leading to DNA damage, which provides a novel theoretical basis for phytomedicine molecular therapy of malignant tumors.

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confidence: 99%

Dihydroartemisinin-driven selective anti-lung cancer proliferation by binding to EGFR and inhibition of NRAS signaling pathway-induced DNA damage

Li,

Peng,

Yang

et al. 2024

Sci Rep

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Comparison of the relative expression of the IGF gene family in the bone marrow of patients with AML compared with normal individuals

Moonesi

Omran

Khosravi

et al. 2023

Preprint

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Introduction: One of the most common types of leukemia is acute myeloid leukemia (AML). Intrinsic and extrinsic factors may lead to AML. Insulin-like growth factor (IGF) is a mitogenic intermediate from the liver that regulates growth and proliferation in response to GH. In this study, we examined the expression of IGF family genes in AML patients (M3 and Non-M3) and compared them with normal samples. Methods Forty bone marrow samples from recently diagnosed AML patients along with 15 samples from subjects without hematological malignancies were collected. For molecular tests, RNA extraction and cDNA synthesis were performed. Finally, IGF1, IGF2, IGFBP3, IGF1R, and IGF2R gene expression were examined by Real-Time PCR. Results IGF1, IGF1R, and IGFBP3 gene expression were significantly increased in patients with AML. In contrast, IGF2 and IGF2R genes did not show significant expression changes between the two groups. Conclusion The expression in this gene family soared in AML patients' bone marrow, compared to normal subjects. This can be caused by malignant cells in the bone marrow. These malignant cells express proteins that increase the number of malignant cells. Moreover, they can be considered as diagnostic biomarkers or therapeutic targets with further research.

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The Effect of DNA Concentration on the HRM Performance in Detecting Jak2 p.V617F Variant in Patients with Myeloproliferative Neoplasms

Moradabadi

Soltani

Shariati

et al. 2023

CST

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Introduction: Janus kinase 2 (JAK2) is an intracellular signaling protein. JAK2 p. V617F is a common variant in normal karyotype myeloproliferative neoplasms (MPN). High-resolution melting (HRM) analysis is one of the essential methods for detecting the JAK2 p.V617F variant. In this study, we have investigated the effect of DNA concentration on detecting the JAK2 p.V617F variant using the HRM method. Materials and Method: Genomic DNA was extracted from human blood and diluted ten times in distilled water from 1 to 0.03; afterward, HRM was conducted for each dilution (triplicate). Using SPSS v.20.0 software, the mean Tm of each dilution was calculated and compared. Results: The HRM results revealed the JAK2 wild type and variant to have Tms of 81/64°C and 80/76°C, respectively. At the endpoint of the pre-amplification, the dilutions had different emissions. The statistical analysis revealed no statistically significant differences in Tm between samples with varying DNA concentrations (P value > 0.05). Conclusion: There have been no significant differences obtained in the analysis of JAK2 p.V617F point variant in different DNA dilutions, implying that the HRM analysis has no relation to DNA concentration.

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Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Cited by 6 publications

References 35 publications

Dihydroartemisinin-driven selective anti-lung cancer proliferation by binding to EGFR and inhibition of NRAS signaling pathway-induced DNA damage

Dihydroartemisinin-driven selective anti-lung cancer proliferation by binding to EGFR and inhibition of NRAS signaling pathway-induced DNA damage

Comparison of the relative expression of the IGF gene family in the bone marrow of patients with AML compared with normal individuals

The Effect of DNA Concentration on the HRM Performance in Detecting Jak2 p.V617F Variant in Patients with Myeloproliferative Neoplasms

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