title: MicroRNA-133b is regulated by TAp63 while no gene mutation is present in colorectal cancer.
abstract: Downregulation of miR-133b has been reported in multiple types of malignancies including colorectal cancer (CRC). We previously confirmed that TAp63 actively translates microRNA-133b (miR-133b) transcripts. While the presence of miRNA mutations have frequently been described in CRC, most CRCs do not show any variation in the miR‑133b coding sequence. Therefore, it is important to elucidate the relationship between TAp63 and miR-133b, and identify other mediators of miR-133b downregulation in CRC. The expression of TAp63 was detected by RT-qPCR, western blotting, immunohistochemistry (IHC) and densitometric analysis using Image-Pro Plus 6.0 software in 38 CRC and corresponding non-cancerous tissues (NCTs). The expression of mature miR‑133b was determined by RT-qPCR, in situ hybridization (ISH) and densitometric analysis using Image-Pro Plus 6.0 software. The DNA from 38 CRC tissues and NCTs were screened for miR-133b mutations through sequence analysis. Compared with the NCTs, TAp63 mRNA expression was significantly lower in 21 (55.27%) tumor tissues. Compared with the NCTs, the miR‑133b expression level was significantly lower in 31 (81.58%) tumor tissues. The expression of miR‑133b was found to be positively correlated with TAp63. Loss of TAp63 and miR-133b was associated with an increased likelihood of metastatic events. The area under the ROC curve (AUC) of TAp63 for CRC was 0.623 [95% confidence interval (CI), 0.497-0.748; P=0.046], with 73.7% sensitivity and 50% specificity, respectively. The AUC of miR-133b for CRC was 0.857 (95% CI, 0.774‑0.940; P<0.0001), with 78.9% sensitivity and 81.6% specificity, respectively. The combined AUC of TAp63 and miR-133b for CRC was 0.881 (95% CI, 0.805-0.956; P<0.0001), with 89.5% sensitivity and 71.1% specificity, respectively. Point mutations within the seed region of miR-133b were found in 1 patient, but the point mutation did not impact the secondary structure of the pre-miR-133b. Therefore, downregulation of TAp63 may be one reason for the dysregulation of miR‑133b in CRC. The expression analysis of TAp63 and miR-133b revealed that they may be used as valuable prognostic biomarkers for CRC.
authors.affiliation: Department of Gastrointestinal Surgery, The Third XiangYa Hospital of Central South University, Changsha, Hunan 410013, P.R. China.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Oncological Surgery, Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221000, P.R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Breast, Thyroid and Vascular Surgery, Chenzhou No. 1 People's Hospital, Chenzhou, Hunan 423000, P.R. China.
authors.forename: Jianhuai
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Gastrointestinal Surgery, The Third XiangYa Hospital of Central South University, Changsha, Hunan 410013, P.R. China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Department of Gastrointestinal Surgery, The Third XiangYa Hospital of Central South University, Changsha, Hunan 410013, P.R. China.
authors.forename: Changwei
authors.initials: C
authors.lastname: Lin
authors.affiliation: Department of Gastrointestinal Surgery, The Third XiangYa Hospital of Central South University, Changsha, Hunan 410013, P.R. China.
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Li
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D008207:Lymphatic Metastasis; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D012372:ROC Curve; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; C548234:MIRN133 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins