miR-638-5p'nin Ekspresyon Artışı PGK1 ve PIM1 Aracılığıyla Akut Miyeloid Lösemide in vitro Hücre Proliferasyonunu Baskılayabilir

Abstract

Amaç: miR-638-5p, Akut Miyeloid Lösemi (AML) dahil olmak üzere çeşitli kanser türlerinde önemli bir tümör baskılayıcı miRNA'dır. Bu çalışma, miR-638-5p'nin ve potansiyel hedef genlerinin HL-60 ve NB4 akut promyelositik lösemi hücre hatları üzerindeki rolünü in vitro ortamda incelemeyi amaçlamaktadır. Metod: miR-638-5p mimik transfeksiyonundan sonra AML hücrelerinin canlılığı üzerindeki etki WST-8 yöntemi ile apoptoz üzerindeki etki ise Caspase-3 kantifikasyon yöntemi ile incelendi. miRWalk, miRDB ve miRTarBase gibi in silico araçlar miR-638-5p'nin olası hedef genlerini seçmek için kullanıldı. Seçilen genlerin ifade düzeyleri qRT-PCR ile araştırıldı. AML hastalarının genel sağkalım (OS) oranı BloodSpot veritabanı üzerinden araştırıldı, zenginleştirme analizi için Enrichr aracı kullanıldı ve korelasyon analizi Correlation AnalyzeR aracı kullanılarak gerçekleştirildi. Bulgular: Kontrollerle karşılaştırıldığında miR-638-5p mimik transfekte hücrelerde azalmış proliferasyon ve artmış apoptoz belirlendi. MECP2, PIM1, MEF2C, PGK1 ve SPAG1 genleri in vitro çalışma için miR-638-5p potansiyel hedefleri olarak seçildi. PGK1 ve PIM1 ekspresyon seviyeleri miR-638-5p mimic transfekte her iki hücrede de önemli ölçüde baskılandı. OS araştırması, MECP2, MEF2C ve PGK1'in aşırı ekspresyonunun AML hastalarının sağkalımını etkilemediğini; ancak SPAG1 ve PIM1'in aşırı ekspresyonunun AML sağkalımı üzerinde zararlı bir etkiye sahip olduğunu ortaya koydu. Ayrıca, zenginleştirme analizi yoluyla PIM1 ve PGK1 genleri arasında pozitif bir korelasyon tespit edildi. Sonuç: miR-638-5p'nin PGK1 ve PIM1 genlerini hedef alarak AML patogenezine katkıda bulunabileceği ve bu durumun AML hücrelerinde hücre proliferasyonunu düzenleyen bir biyomolekül olarak potansiyeline işaret edebileceği düşünülmektedir.

Keywords

• miR-638-5p
• Akut Miyeloid Lösemi
• HL-60
• NB4
• mikroRNA mimik transfeksiyonu

Enhanced Expression of miR-638-5p May Suppress Acute Myeloid Leukemia in vitro Cell Proliferation Through PGK1 and PIM1

Abstract

Objective: miR-638-5p is a crucial tumor suppressor miRNA in several cancer types including, Acute Myeloid Leukemia (AML). This study aimed to analyze the role of miR-638-5p and its potential target genes in HL-60 and NB4 acute promyelocytic leukemia cell lines using in vitro method. Method: After the miR-638-5p mimic transfection into AML cells, the effect on cell viability was examined by the WST-8 method, and the effect on apoptosis was measured via the Caspase-3 quantification method. In silico tools such as miRWalk, miRDB, and miRTarBase were used to select the possible target genes of miR-638-5p. The expression levels of selected genes were investigated by qRT-PCR. The overall survival (OS) rate of AML patients was explored via the BloodSpot database, the Enrichr tool was used for enrichment analysis, and correlation analysis was performed using the Correlation AnalyzeR tool. Results: Decreased proliferation and increased apoptosis were determined in miR-638-5p mimic transfected cells compared to the controls. MECP2, PIM1, MEF2C, PGK1, and SPAG1 genes were selected as the potential targets of miR-638-5p for in vitro study. PGK1 and PIM1 expression levels were significantly suppressed in cells transfected with the miR-638-5p mimic. The OS investigation revealed that overexpression of MECP2, MEF2C, and PGK1 does not affect the survival of AML patients; however, overexpression of SPAG1 and PIM1 has a detrimental effect on AML survival. Also, a positive correlation was detected between PIM1 and PGK1 genes via enrichment analysis. Conclusion: miR-638-5p may contribute to AML pathogenesis by targeting the PGK1 and PIM1 genes, and this situation may indicate its potential as a biomolecule for regulating cell proliferation in AML cells.

Keywords

• miR-638-5p
• Acute Myeloid Leukemia
• HL-60
• NB4
• microRNA mimic transfection

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