miR-142-3p simultaneously targets HMGA1, HMGA2, HMGB1, and HMGB3 and inhibits tumorigenic properties and in-vivo metastatic potential of human cervical cancer cells

• Published in: Life sciences (1973) Vol. 291; p. 120268
• Main Authors: Sharma, Priyanshu, Yadav, Poonam, Jain, Ruchi P., Bera, Amal Kanti, Karunagaran, Devarajan
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 15.02.2022; Elsevier BV
• Subjects: 3' Untranslated regions; Animals; Annexin V; Apoptosis; Cancer; Carcinogenesis - genetics; Cell death; Cell Line, Tumor; Cell migration; Cell Movement - genetics; Cell proliferation; Cell Proliferation - drug effects; Cell viability; Cervical cancer; Cervix; Chemical compounds; Colonies; Danio rerio; Female; Flow cytometry; Gene Expression - genetics; Gene Expression Regulation, Neoplastic - genetics; Gene targeting; HeLa Cells; High-mobility-group proteins; HMGA1a Protein - genetics; HMGA1a Protein - metabolism; HMGA2 Protein - genetics; HMGA2 Protein - metabolism; HMGB1 protein; HMGB1 Protein - genetics; HMGB1 Protein - metabolism; HMGB3 Protein - genetics; HMGB3 Protein - metabolism; Humans; In vivo methods and tests; Metastases; Metastasis; MicroRNAs - genetics; MicroRNAs - metabolism; miR-142-3p; miRNA; Models, Animal; Neoplasm Invasiveness - genetics; NF-κB; Oncogenes; Pharmacology; Proteins; siRNA; Tumor cell lines; Tumors; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - metabolism; Xenograft Model Antitumor Assays; Zebrafish; Zebrafish tumor xenograft; NF-κB; High-mobility-group proteins; miR-142-3p; Metastasis; Zebrafish tumor xenograft; Apoptosis
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2021.120268

High-mobility group (HMG) proteins are oncogenic in different cancers, including cervical cancer; silencing their individual expression using sh-RNAs, siRNAs, and miRNAs has had anti-tumorigenic effects, but the consequences of their collective downregulation are not known. Since multiple gene targeting is generally very effective in cancer therapy, the present study highlighted the consequences of silencing the expression of HMGA1, A2, B1, and B3 using sh-RNAs or miR-142-3p (that can potentially target HMGA1, A2, B1, and B3) in cervical cancer cell lines. 3′ UTR luciferase reporter assays were performed to validate HMGA1, A2, B1, and B3 as targets of miR-142-3p in human cervical cancer cells. Annexin V/PI dual staining and flow cytometry analyses were used to detect apoptotic cells. miR-142-3p-mediated regulation of cell death, colony formation, migration, and invasion was investigated in human cervical cancer cells together with in vivo metastasis in zebrafish. Concurrent knockdown of HMGA1, A2, B1, and B3 through their corresponding sh-RNAs inhibited cell viability and colony formation but induced apoptosis, and these effects were relatively reduced upon their individual knockdown. miR-142-3p targeted HMGA1, A2, B1, and B3 by binding to their 3′UTRs and induced apoptosis but inhibited proliferation, migration, and invasion of human cervical cancer cells. In addition, miR-142-3p expression decreased phospho-p65 and EMT-related proteins in cervical cancer cells and their in vivo metastatic potential upon implantation in zebrafish. These findings suggest that miR-142-3p acts as a tumor-suppressive miRNA by targeting HMGA1, A2, B1, and B3 and may serve as a potential therapeutic agent in human cervical cancer. •Concurrent inhibition of HMGA1, HMGA2, HMGB1 and HMGB3 exerts anti-tumor effects in human cervical cancer cells•shRNA-mediated knock-down of HMG genes upregulates miR-142-3p levels•miR-142-3p can target HMGA1, A2, B1 and B3 directly by binding to their 3’UTR in cervical cancer cells•miR-142-3p mediated anti-tumorigenic effects are routed via HMG genes as their restoration partially rescues these effects•miR-142-3p inhibits metastasis in vivo (zebrafish tumor xenograft), EMT and NF-κB signaling