# 14-3-3β knockdown inhibits migration and colony formation of esophageal squamous cell carcinoma cells and is associated with weakened p-AKT signaling

## TL;DR

This study shows that reducing 14-3-3β in esophageal cancer cells slows their movement and growth, possibly by weakening the p-AKT signaling pathway.

## Contribution

The study reveals a novel link between 14-3-3β and p-AKT signaling in ESCC progression.

## Key findings

- 14-3-3β knockdown in KYSE-150 cells reduced migration and colony formation.
- Reduced 14-3-3β expression led to decreased p-AKT activity without affecting AKT levels.
- No significant changes in apoptosis-related proteins were observed after 14-3-3β knockdown.

## Abstract

This study aims to explore the functions and mechanisms underlying the involvement of 14-3-3β in esophageal squamous cell carcinoma (ESCC).

Western blot and quantitative real-time PCR (RT-PCR) were utilized to evaluate the levels of 14-3-3β in SHEE, TE-1, and KYSE-150 cells. Following infection with YWHAB-RNAi lentivirus and puromycin screening, the reduction of 14-3-3β expression in KYSE-150 cells was confirmed via western blot and RT-PCR. Subsequently, wound healing, CCK8, and colony-formation assays were performed to assess cell migration and proliferation in KYSE-150 cells with stable low levels of 14-3-3β. Furthermore, western blot analysis was conducted to examine the expression of relevant proteins, elucidating the potential molecular mechanisms underlying 14-3-3β involvement in ESCC progression and metastasis.

The expression of 14-3-3β was significantly elevated in KYSE-150 cells compared to SHEE cells. Upon infection in KYSE-150 cells, the expression of 14-3-3β was diminished, resulting in suppressed cell migration and colony formation, although no significant changes were observed in the CCK8 assays. Furthermore, in KYSE-150 cells exhibiting reduced 14-3-3β expression, the activity of phosphorylated AKT (p-AKT) was significantly decreased, while no notable difference was detected in AKT protein levels. Additionally, there were no significant alterations observed in apoptosis-related proteins, including BCL2, BAX, Caspase3, and activated Caspase9.

In KYSE-150 cells infected with YWHAB-RNAi lentivirus, the downregulation of 14-3-3β expression resulted in significant reductions in both cell migration and proliferation. Furthermore, the findings demonstrated that 14-3-3β may regulate the biological activities of ESCC cells via modulation of the PI3K/AKT signaling pathway.

The online version contains supplementary material available at 10.1186/s41065-026-00651-4.