RNAi-mediated IARS2 knockdown inhibits proliferation and promotes apoptosis in human melanoma A375 cells

Abstract

IARS2, which encodes the mitochondrial form of isoleucyl-tRNA synthetase, has been found to play an important role in a range of diseases, including cancer. However, the relationship between IARS2 and melanoma is still unclear. To evaluate the role of IARS2 in melanoma, we constructed a stable A375 cell line with IARS2 knockdown via lentivirus-mediated small interfering RNAs. The expression of IARS2 was measured by real time-quantitative Polymerase Chain Reaction and western blot analysis. Cell counting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and colony formation assay were conducted to assess the effect of IARS2 on melanoma cell proliferation. Flow cytometry assay was used to determine cell apoptosis and cell cycle distribution in melanoma A375 cells. Finally, immunohistochemistry was employed to validate the expression of IARS2 protein in melanoma tissues. In this study it was found that IARS2 was highly expressed in melanoma cell lines. Furthermore, IARS2 protein also exhibited elevated expression in the tumour tissues obtained from melanoma patients. After suppression of the mRNA expression of IARS2, the proliferation and colony formation ability of the A375 cells were significantly inhibited, while the proportion of apoptotic A375 cells increased significantly, as indicated by an enhanced phosphatidylserine externalization and caspase 3/7 activity after IARS2 knockdown. Further investigations found that knockdown of IARS2 arrested cells in the G1 phase. The results suggested that IARS2 is critical for proliferation and apoptosis of melanoma cells.

Lentivirus‑induced knockdown of IARS2 expression inhibits the proliferation and promotes the apoptosis of human osteosarcoma cells

“…The present study was, to the best of our knowledge, the first to demonstrate that IARS2 knockdown inhibited cell proliferation and promoted the apoptosis of human OS cells, which is consistent with previous studies on the role in IARS2 in other tumor types ( 19 – 21 ). In the present study, the expression of IARS2, and its relationship with overall and disease-free survival were analyzed using the GEPIA2 database.…”

“…Thus, whether these signaling pathways are also involved in OS cell proliferation and apoptosis driven by IARS2 should be analyzed in future research. Furthermore, due to the normal sample limitation in the GEPIA2 dataset and previous study has not reported an association between survival and IARS2 expression so far ( 19 – 21 ), the expression and survival curve should be further confirmed with more healthy and OS clinical samples. In addition, the expression of IARS2 in tumor and tumor-adjacent tissue needs to be detected by western blotting and immunohistochemistry.…”

“…A number of studies have revealed that IARS2 is associated with various types of cancer. IARS2 knockdown has been shown to inhibit the proliferation and promote the apoptosis of human A375 melanoma cells and AGS GC cells ( 16 , 21 ). In acute myeloid leukemia (AML) cells, knockdown of IARS2 can also inhibit proliferation, which is achieved by regulating the p53/p21/PCNA/Eif4E pathway ( 19 ).…”

Identification of a novel lactylation-related gene signature predicts the prognosis of multiple myeloma and experiment verification

“…IARS2 mainly catalyzes tRNA aminoacylation and correlates with malignant proliferation and apoptosis resistance of tumor cells. In non-small cell lung cancer, gastric cancer, melanoma and acute myeloid leukemia, knockout of IARS2 inhibits cell proliferation and promotes apoptosis 36 – 39 . PRCC is an important element involved in the formation of the spliceosome 40 .…”

New Insights Into Human Hyaluronidase 4/Chondroitin Sulphate Hydrolase

“…IARS2 is a ubiquitously expressed mitochondrial tRNA synthase that catalyses the aminoacylation of tRNA with isoleucine. Knockdown of IARS2 has been shown to promote apoptosis and inhibit proliferation in melanoma cells ( Ma et al, 2020 ). NEK4 is a serine/threonine kinase involved in replicative senescence and for normal cell cycle arrest in response to double-stranded DNA damage ( Nguyen et al, 2012 ).…”