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[The Regulatory Effect of RNA m6 A Methylation Modification on KDM4B Gene Expression in t (8;21) AML Cells by MeRIP-qPCR].
Li, Yu-Qing; Shao, Yang-Liu; Li, Meng-Yue; Wang, Li-Li; Gao, Xiao-Ning.
Afiliación
  • Li YQ; Department of Hematology, The Fifth Medical Center, Chinese PLA General Hospital, Beijing 100071, China.
  • Shao YL; Medical School of Chinese PLA, Beijing 100853, China.
  • Li MY; Department of Hematology, The Fifth Medical Center, Chinese PLA General Hospital, Beijing 100071, China.
  • Wang LL; Medical School of Chinese PLA, Beijing 100853, China.
  • Gao XN; Department of Hematology, The Fifth Medical Center, Chinese PLA General Hospital, Beijing 100071, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 32(2): 382-388, 2024 Apr.
Article en Zh | MEDLINE | ID: mdl-38660840
ABSTRACT

OBJECTIVE:

To confirm the direct regulatory effect of WTAP-mediated RNA m6A modification on the KDM4B gene in t (8;21) acute myeloid leukemia (AML) cells through MeRIP combined with reverse transcription real-time quantitative PCR (RT-qPCR) technology.

METHODS:

The lentivirus-mediated shRNA target WTAP or KDM4B gene was used to transfect the t (8;21) AML cell lines Kasumi-1 and SKNO-1, and cells transfected with randomly shuffled shRNA as the control. Using the Ultrapure RNA Extraction Kit (DNase I) to extract RNA. The Magna MeRIPTM m6A Kit was used to enrich methylated modified fragments, and detect the m6A methylated RNA regions by RT-qPCR, and the protein and mRNA expression levels of WTAP and KDM4B in cells were detected by Western blot and reverse transcription real-time quantitative PCR (RT-qPCR). Colony formation assays were used to detect the colony ability of cells in vitro.

RESULTS:

Silencing the expression of WTAP in Kasumi-1 cells, the enrichment of m6A methylation modification was significantly decreased in the 3'UTR of KDM4B mRNA(P < 0.01), and the protein(P < 0.001) and mRNA (Kasumi-1P < 0.001; SKNO-1 P < 0.01) expression levels of KDM4B were also significantly inhibited in Kasumi-1 and SKNO-1 cells upon WTAP knockdown (all P < 0.01), accompanied by a significant decrease in the colony-forming ability of both cell lines (both P < 0.01).

CONCLUSION:

In t(8;21) AML cell lines, WTAP could regulate the expression of KDM4B by regulating the m6A modification of the 3'UTR of KDM4B mRNA, and silencing the expression of KDM4B could inhibit the cellular proliferation in vitro.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Leucemia Mieloide Aguda / Histona Demetilasas con Dominio de Jumonji Límite: Humans Idioma: Zh Revista: Zhongguo Shi Yan Xue Ye Xue Za Zhi Asunto de la revista: HEMATOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: China
Texto completo
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XML
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Leucemia Mieloide Aguda / Histona Demetilasas con Dominio de Jumonji Límite: Humans Idioma: Zh Revista: Zhongguo Shi Yan Xue Ye Xue Za Zhi Asunto de la revista: HEMATOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: China