Potential Roles of IGF2BP3 in Bladder Cancer [m6A-seq]

TCGA data analysis and clinical sample test results showed the IGF2BP3 is highly expressed in bladder cancer and related to tumor metastasis and invasion. RNA sequencing results of clinical bladder tissue from 53 cases showed that the IGF2BP3 expression is related to cell proliferation genes. M6a and RNA sequencing results showed the IGF2BP3 affects the growth, proliferation, and apoptosis of T24 cells, and SpHK1, POMT2, and MRPS18a may be potential targets of IGF2BP3. Overall design: (1) The TCGA bladder cancer gene expression and the survival data were used to screen the m6A-related RNA-binding protein IGF2BP3. (2) The expression level of the IGF2BP3 in the bladder tissue was detected using immunohistochemistry and reverse transcription-quantitative PCR . (3) The CRISPR/Cas9 was used to construct a T24 stable cell line with the IGF2BP3 knockout, and DNA sequencing and Western blot were used to verify the knockout effect. (4) The MTT test, colony-forming assay, flow cytometry, scratch test, and transwell invasion test were used to detect the changes in the biological function in IGF2BP3 knockout T24 cell lines. (5) The RNA was extracted from the bladder tissue for sequencing to analyze the role of IGF2BP in bladder cancer. (6) The gene expression and the M6a level in T24 knockout IGF2BP3 cells were analyzed using RNA-seq and Merip-seq, and the mechanism of IGF2BP3 in bladder cancer was preliminarily discussed.