EXPERIMENTAL AND THERAPEUTIC MEDICINE - 26 - Malignant biological behaviors of oral squamous cell carcinoma via CIP2A/AKT1 and the GSK-3β/β-catenin pathway

Sep 24, 2026

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Oral squamous cell carcinoma (OSCC) is a prevalent malignancy with poor prognosis, and this study investigates the role of cancerous inhibitor of protein phosphatase 2A (CIP2A) and its regulatory effect on AKT1. CIP2A and AKT1 levels in OSCC cells were measured by western blotting; CIP2A knockdown assessed viability, proliferation, apoptosis, migration, invasion, and tube formation through CCK-8, EdU, TUNEL, wound healing, Transwell, and co-culture assays. Co-immunoprecipitation verified CIP2A–AKT1 interaction. AKT1 overexpression rescued malignant phenotypes and restored phosphorylated GSK-3β and β-catenin signaling, supporting targeted therapeutic potential for OSCC.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages11

Reading Time28 min

AccessFree to download

Abstract

Experimental aims and rationale

Methods: expression detection and functional assays

Interaction validation and rescue experiments

Key results and conclusion

EXPERIMENTAL AND THERAPEUTIC MEDICINE 26: 514, 2023 CIP2A interacts with AKT1 to promote the malignant biological behaviors of oral squamous cell carcinoma by upregulating the GSK‑3β/β‑catenin pathway YILEI CHE, HUI ZHANG, HUI LI and XIAOZHEN WU Department of Stomatology, Aerospace Center Hospital, Beijing 100049, P.R. China Received May 16, 2023; Accepted August 18, 2023 DOI: 10. 3892/etm.2023.12213 Abstract. Oral squamous cell carcinoma (OSCC) is one of the most common malignancies worldwide, which is asso‑ ciated with a poor prognosis . The present study aimed to investigate the role of cancerous inhibitor of protein phos‑ phatase 2A (CIP2A) in OSCC and its regulatory effect on AKT1 . Firstly, CIP2A and AKT1 expression in OSCC cells was detected by western blotting. After silencing CIP2A, cell viability and cell proliferation were assessed using the Cell Counting Kit‑8 assay and 5‑ethynyl‑2'‑deoxyuridine staining. Cell apoptosis was evaluated by TUNEL staining and the expression of apoptosis‑related proteins was assessed using western blotting. Wound healing, Transwell and tube formation assays were performed to evaluate CAL‑27 cell migration, invasion and human umbilical vein endothelial cell (HUVEC) tube formation . The interaction between CIP2A and AKT1 was identified by co‑immunoprecip‑ itation (co‑IP) . In addition, AKT1 was overexpressed in CIP2A‑silenced CAL‑27 cells to perform rescue experiments to analyze the malignant biological functions of CAL‑27 cells . Finally, the expression of proteins in the glycogen synthase kinase (GSK)‑3β/β‑catenin pathway was deter‑ mined by western blot analysis . Markedly elevated CIP2A and AKT1 expression was observed in OSCC cells. CIP2Aknockdown inhibited the viability, proliferation, migration and invasion, and promoted the apoptosis of CAL‑27 cells. Concurrently, CIP2A loss‑of‑function attenuated tube forma‑ tion . Results of Co‑IP confirmed there was an interaction between CIP2A and AKT1 . Rescue experiments suggested that AKT1 overexpression alleviated the inhibitory effects of CIP2A knockdown on the viability, proliferation, migra‑ tion and invasion of CAL‑27 cells, as well as tube formation Correspondence to: Professor Xiaozhen Wu, Department of Stomatology, Aerospace Center Hospital, 15 Yuquan Road, Haidian, Beijing 100049, P.R. China E‑mail: [xiaozw860324@163.com](xiaozw860324@163.com) Key words: oral squamous cell carcinoma, cancerous inhibitor of protein phosphatase 2A, AKT1, migration, invasion, GSK‑3β/β‑catenin in HUVECs . Additionally, CIP2A silencing significantly downregulated phosphorylated‑GSK‑3β and β‑catenin expression, which was reversed by AKT1 overexpression. In conclusion, CIP2A could interact with AKT1 to promote the malignant biological behaviors of OSCC cells by upregu‑ lating the GSK‑3β/β‑catenin pathway. These findings may provide a targeted therapy for OSCC treatment. Introduction Oral squamous cell carcinoma (OSCC) represents the most frequent form of head and neck squamous cell carcinoma, accounting for 90% of all oral cancer cases worldwide. In addition, OSCC is associated with a poor prognosis with a 5‑year overall survival rate of only ~50% globally (1,2) . A previous study reported that >30% of patients with OSCC may develop multiple tumors within 5‑10 years, which further aggravates this prognosis (3) . Although great prog‑ ress has been made in surgical techniques, radiotherapy and chemotherapy, the survival rate remains relatively low, due to the local recurrence and metastasis of the disease (4,5) . Therefore, an in‑depth understanding of the molecular mechanism underlying the occurrence and development of OSCC is of crucial importance for the development of new therapeutic strategies. Cancerous inhibitor of protein phosphatase 2A (CIP2A), originally named KIAA1524 or p90, has been reported to act as an oncogene by promoting tumor growth in several types of cancer, including bladder cancer, non‑small cell lung cancer

FAQs

What is the main objective of the study on CIP2A in OSCC?

The study aims to determine how CIP2A influences OSCC malignant behaviors and how it regulates AKT1.

How did the researchers assess CIP2A’s effects on OSCC cell behavior?

They used western blotting to detect CIP2A and AKT1, then performed CCK-8 and EdU for viability/proliferation, TUNEL and apoptosis-related proteins for apoptosis, and wound healing and Transwell assays for migration and invasion. Tube formation assays evaluated angiogenesis-related effects.

What evidence supports an interaction between CIP2A and AKT1?

Co-immunoprecipitation (co-IP) confirmed that CIP2A interacts with AKT1.

How do AKT1 overexpression experiments affect the impact of CIP2A knockdown?

AKT1 overexpression alleviated the inhibitory effects of CIP2A knockdown on viability, proliferation, migration, invasion, and tube formation, and reversed changes in the phosphorylated GSK-3β/β-catenin pathway.

Sep 24, 2026

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