ALKBH1 contributes to renal cell carcinoma progression by reducing N6‐methyladenine of GPR137

Abstract

Background Renal cell carcinoma (RCC) accounts for approximately 4% of all adult malignancies with high mortality worldwide. Although conventional chemotherapy and radiotherapy treatment has been applied for RCC in clinic, the mortality rate of patients is increasing each year, and patients with metastatic RCC are still suffering from poor prognosis. Thus, further investigation of the molecular mechanisms responsible for the development and progression of RCC is of particular importance. Methods Total of 10 pairs of RCC tissues and adjacent nontumor tissues were collected for examination of ALKBH1 and GPR137 expression. The correlations between ALKBH1 and GPR137 expression in RCC patient were assessed by GEPIA online tool and were analyzed using auto best cutoff. The human RCC cell lines Caki‐1, 786‐O, ACHN, Osrc2, A498, and 769‐P, were used for mechanistic investigation. Results Here, we report that the expression of AlkB homologue 1 (ALKBH1) is upregulated in RCC tissues, which is correlated with G‐protein‐coupled receptor 137 (GPR137) expression. The elevated expression of ALKBH1 is associated with RCC cell malignant characteristics, including cell proliferation and movement (migration and invasion). Mechanistic investigation further reveals that ALKBH1 reduces m6A levels of GPR137 mRNA in RCC cells, which upregulates GPR137 mRNA levels, resulting in the increased GPR137 protein expression subsequently and the enhanced RCC cell biological actions consequently. In contrast, the suppression of GPR137 effectively alleviates the ALKBH1‐induced malignancies of RCC cells. Conclusion Our results indicate that ALKBH1–GPR137 axis might be used as a potential therapeutic target in RCC, contributing to finding new prognostic biomarkers for RCC at an early stage.

GPR137 inactivates Hippo signaling to promote gastric cancer cell malignancy

“…Slides were analyzed by a laser scanning microscope (Zeiss, Germany). GPR137 expression was scored as previously described [ 18 ] based on the proportion of cells showing GPR137 immunostaining across 3 non-adjacent fields in each sample with the following criteria: 0 (no cell positive); 1 (< 50% cells weakly positive); 2 (< 50% cells intensely staining); 3 (≥ 50% cells weakly positive); 4 (≥ 50% cells intensely staining).…”

PUS1 is a novel biomarker for evaluating malignancy of human renal cell carcinoma

“…CCK-8 assays were conducted as described previously [ 26 , 27 ]. Briefly, cells were seeded into 96-well plates and were infected with PUS1-expressing lentiviruses, control lentiviruses, PUS1-shRNA-expressing lentiviruses, or lentiviruses with scrambled shRNA.…”

“…Total protein from human RCC Caki-1, 786-O, ACHN, OSRC2, A498, Caki-2 and 769P cells were obtained using RIPA lysis buffer (Beyotime, Shanghai, China), and protein concentrations were determined by standard Bradford assay with a commercial kit (Beyotime, Shanghai, China), followed by western blotting as described previously [24][25][26]. Briefly, 50 µg of total protein was subjected to SDS-PAGE followed by a transfer onto polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA, USA).…”

The role of RNA-modifying proteins in renal cell carcinoma

“…A similar oncogenic effect was detected in ALKBH3 in RCC patients, in which its expression was positively correlated with advanced TNM staging and poor prognosis [ 94 ]. Moreover, ALKBH1 had an oncogenic role in RCC cell lines and promoted cell migration and viability [ 95 ]. Overall, further studies are required to elucidate the molecular mechanisms behind the different roles of ALKBH in RCC.…”