WTAP promotes proliferation and aerobic glycolysis via regulating m6A modification of BMI1 mRNA

doi:10.19401/j.cnki.1007-3639.2023.07.003

Abstract

Abstract:

Background and purpose: Methyltransferase Wilms’ tumor 1-associating protein (WTAP) has been proven to play an important role in the development of tumors. This study aimed to explore the influence of WTAP on the occurrence and development of glioma from the perspective of N6-methyladenosine (m6A) modification. Methods: The differentially expressed m6A regulators in glioma were screened from GEO database. The expression of WTAP/B cell-specific Moloney murine leukemia virus integration site 1 (BMI1) in glioma tissue and cell lines was evaluated by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Methyl thiazoyl terazolium (MTT) was used to explore the effects of WTAP/BMI1 on glioma cell proliferation. Extracellular acidification rate (ECAR) kits and glucose consumption and lactate production detection kits were utilized to detect aerobic glycolysis of glioma cells. Results: Compared with normal tissues, WTQP and BMI1 expressions were upregulated in glioma tissues. Otherwise, compared with WTAP (1.01±0.13) and BMI1 (1.02±0.11) expressions in HEB cells, WTAP (2.38±0.17) and BMI1 (2.25±0.14) expressions in U251 cells were increased (P<0.05). Downregulation of WTAP inhibited cell proliferation and glycolytic potential in vitro, while overexpression of WTAP showed opposite effects. The effects of WTAP overexpression on glioma cell proliferation and glycolysis were partially rescued by BMI1 inhibition (P<0.05). WTAP promoted m6A methylation modification of BMI1 and enhanced the stability of BMI1 mRNA (P<0.05). Conclusion: WTAP upregulates the level of BMI1 in a m6A methylation modification dependent manner, subsequently promotes glioma cell proliferation and aerobic glycolysis. Molecular therapy targeting WTAP may provide a new method for the treatment of glioma.

Key words: Glioma, N6-methyladenosine modification, Aerobic glycolysis, Methyltransferase

CLC Number:

R739.41

DIAO Xinfeng, LI Xinmao, HOU Liang, WEI Zhixuan. WTAP promotes proliferation and aerobic glycolysis via regulating m6A modification of BMI1 mRNA[J]. China Oncology, 2023, 33(7): 655-663.

Figures/Tables 8

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Gene	Forward primer (5‘-3‘)	Reverse primer (5‘-3‘)
WTAP	GCGACTAGCAACCAAGGAAC	CATTTTGGGCTTGTTCCAGT
BMI1	CCAGGGCTTTTCAAAAATGA	CCGATCCAATCTGTTCTGGT
GAPDH	CGACCACTTTGTCAAGCTCA	AGGGGTCTACATGGCAACTG

Clinical data	Case n (n=47)	WTAP expression		P value
Clinical data	Case n (n=47)	High expression n	Low expression n	P value
Gender				0.920
Male	25	14	11
Female	22	12	10
Age/year				0.927
≤45	16	9	7
>45	31	17	14
Tumor size/cm				0.474
<3	20	9	11
≥3	27	15	12
WHO grade				0.014 5
Ⅰ-Ⅱ	31	11	20
Ⅲ-Ⅳ	16	12	4

Gene	RF classifier	SVM classifier
MYB	0.65	0.96
APC	0.70	0.98
TP53	0.70	0.79
EGFR	0.80	0.94
BMI1	0.80	0.97
PPM1D	0.65	0.95
ALK	0.80	0.96
NTRK1	0.75	0.59
HLA-A	0.85	0.20