DOI: 10.5061/dryad.b5mkkwhrt
title: Data from: MFSD12, transcriptionally regulated by PLAGL2, promotes bladder cancer progression

description: Bladder cancer (BLCA) is one of the most common malignant tumors of the
 urinary system. Identification of novel molecular signaling targets for
 the tumorigenesis of BLCA is important. Data from the Gene Expression
 Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases suggest that
 major facilitator superfamily domain containing 12 (MFSD12) may act as an
 important oncogene in BLCA. MFSD12 expression is confirmed to be elevated
 in BLCA patients. Genetic manipulation of MFSD12 mediated by Tet-inducible
 lentiviral expression vector is conducted in two BLCA cell lines,
 including UMUC3 and 5637. Following this manipulation, the cells are
 subjected to treatment with or without doxycycline. Our results show that
 MFSD12 knockdown inhibits cell proliferation, migration, and invasion, and
 arrests the G1 stage-induced cell cycle. Furthermore, silencing of MFSD12
 reduces lung metastatic lesions and xenografted tumor formation of BLCA
 cells. To further explore the effect of MFSD12 on BLCA cells,
 transcriptomics and metabolomics analyses are performed on
 MFSD12-overexpressing cells. Subsequently, luciferase reporters and
 chromatin immunoprecipitation (ChIP)-PCR assays reveal that MFSD12 is
 regulated positively by pleomorphic adenoma gene like-2 (PLAGL2), an
 important transcription factor. Collectively, our results indicate that
 MFSD12 exerts a tumor-promoting effect on BLCA progression, under the
 modulation of transcription factor PLAGL2.
description: For transcriptome analysis, UMUC3 cells infected with lentivirus
 overexpressing MFSD12 were treated with 1 μg/mL doxycycline (DOX) for 48
 hours. RNA was extracted utilizing established protocols, and
 complementary DNA (cDNA) libraries underwent high-throughput sequencing
 via the Illumina Sequencing platform. Subsequently, these libraries were
 aligned to the GRCh38 reference genome, as provided by the Ensembl
 database, employing HISAT2 (version 2.2.1). The HTSeq software (version
 2.0.2) was utilized to quantify read counts for each transcript. To
 identify differentially expressed genes (DEGs), DEseq2 software (version
 1.28.0) was applied. For metabonomics, metabolites were
 extracted by adding 1 mL of a solvent mixture comprising acetonitrile,
 methanol, and water in a volumetric ratio of 2:2:1. Subsequently, the
 metabolites were analyzed using a Thermo Q Exactive mass spectrometer in
 conjunction with a Thermo Vanquish ultra-high performance liquid
 chromatography system. The Proteowizard4 software (version 3.0.8789)
 facilitated the conversion of RAW files into .mzXML format, while XCMS
 software (version 3.12.0) was employed for peak alignment, filtering, and
 filling. Metabolite identification was conducted using various public
 databases. Orthogonal partial least squares-discriminant analysis
 (OPLS-DA) was performed using the R "ropls" package (version
 1.30.0), with differentially expressed metabolites (DEMs) identified based
 on a significance threshold of p &lt; 0.05 and a variable importance
 in projection (VIP) score greater than 1. 
description: # Data from: MFSD12, transcriptionally regulated by PLAGL2, promotes
 bladder cancer progression Dataset DOI:
 [10.5061/dryad.b5mkkwhrt](10.5061/dryad.b5mkkwhrt) ## Description of the
 data and file structure ### Files and variables #### File:
 Metabolomics.xlsx **Description:** Metabolomic analysis of UMUC3 cells
 with MFSD12 overexpression File 1 is an Excel file containing
 differentially expressed metabolites in four samples per group, comparing
 Lv-MFSD12 + DOX with Lv-MFSD12. The columns include the metabolite name,
 description, m/z (Mass-to-Charge Ratio), rt (retention time), ppm (parts
 per million), formula, log2 fold change, p-value, and VIP (variable
 importance in projection). The differentially expressed metabolites were
 identified based on VIP score greater than 1 and a significance criterion
 of p &lt; 0.05. ##### Variables: **MFSD12 overexpression or not** ####
 File: Transcriptomics.xlsx **Description:** The Transcriptomic analysis of
 UMUC3 cellswith MFSD12 overexpression File 2 is an Excel file containing
 differentially expressed genes in six samples per group, comparing
 Lv-MFSD12 + DOX vs. Lv-MFSD12. The columns include the Gene Name,
 Description, Ensembl ID, log2FoldChange, p-value, adjust p-value, and
 regulation. The differentially expressed genes were identified based on
 log2 FoldChange greater than 1 and a significance criterion of p &lt;
 0.05. ##### Variables: MFSD12 overexpression or not ## Code/software
 Utilize Microsoft Excel to access and examine files.