ALKBH5 Suppresses Autophagy in Prostate Cancer Cells via Inhibiting m6A-Modification of TSPAN1 mRNA.

Abstract

BACKGROUND:

Activating autophagy promotes the invasion and progression of prostate cancer (PCa). Tetraspanin 1 (TSPAN1) has been found to promote autophagy flux and its up-regulation can enhance the migration of PCa cells. In addition, there is a binding relationship between TSPAN1 and the N6-methyladenosine (m6A) demethylase AlkB homolog 5 (ALKBH5). Therefore, we wanted to know whether ALKBH5 could affect autophagy by regulating TSPAN1 expression, and thereby participate in PCa malignant progression.

METHODS:

The expression of ALKBH5 and TSPAN1 in PCa was examined by quantitative real-time polymerase chain reaction (qRT-PCR), and the functional tests included cell counting kit-8 and 5-ethynyl-2'-deoxyuridine (EdU) staining assays. The expression of autophagy-related proteins was confirmed by western blot. Detection of the m6A level of TSPAN1 was performed using methylated RNA immunoprecipitation sequencing (MeRIP)-qPCR.

RESULTS:

ALKBH5 was significantly downregulated in PCa cells (LNCaP, DU145 and PC3 cells; p < 0.001). Overexpression of ALKBH5 inhibited cell viability and the number of EdU-positive cells (p < 0.01, p < 0.001), decreased the ratio of microtubule-associated protein light chain 3B (LC3B)-II/LC3B-I, and promoted P62 protein expression in LNCaP and DU145 cells (p < 0.001). The m6A level of TSPAN1 was high in LNCaP and DU145 cells, but was inhibited by the overexpression of ALKBH5 (p < 0.001). TSPAN1 overexpression promoted cell viability (p < 0.001), increased EdU-positive cells and the LC3B-II/LC3B-I ratio (p < 0.001, p < 0.05), reduced P62 protein expression (p < 0.05, p < 0.001), and reversed the regulation of ALKBH5 overexpression in LNCaP and DU145 cells (p < 0.01, p < 0.001).

CONCLUSIONS:

Promoting ALKBH5 expression may inhibit PCa autophagy by reducing the m6A level of TSPAN1.

• Authors: Xudong Zhao, Zhiqiang Zhao, Honglin Xie, Wenjun Gao, Chunhua Ye
• Journal: Discovery medicine (Discov Med) Vol. 36 Issue 180 Pg. 61-70 (Jan 2024) ISSN: 1944-7930 [Electronic] United States
• PMID: 38273746 (Publication Type: Journal Article)

Chemical References

• 6-methyladenine
• TSPAN1 protein, human
• Tetraspanins
• ALKBH5 protein, human
• AlkB Homolog 5, RNA Demethylase
• Adenine

Topics

• Male
• Humans
• Prostatic Neoplasms (genetics)
• Prostate
• Autophagy (genetics)
• Cell Survival
• Tetraspanins (genetics)
• AlkB Homolog 5, RNA Demethylase (genetics)
• Adenine (analogs & derivatives)