PIK3CA mutation confers resistance to chemotherapy in triple-negative breast cancer by inhibiting apoptosis and activating the PI3K/AKT/mTOR signaling pathway

Huayu Hu; Junyong Zhu; Yuting Zhong; Rui Geng; Yashuang Ji; Qingyu Guan; Chenyan Hong; Yufan Wei; Ningning Min; Aiying Qi; Yanjun Zhang; Xiru Li

doi:10.21037/atm-21-698

PIK3CA mutation confers resistance to chemotherapy in triple-negative breast cancer by inhibiting apoptosis and activating the PI3K/AKT/mTOR signaling pathway

Ann Transl Med. 2021 Mar;9(5):410. doi: 10.21037/atm-21-698.

Authors

Huayu Hu^{1

2}, Junyong Zhu^{2

3}, Yuting Zhong^{2

3}, Rui Geng^{2

3}, Yashuang Ji^{1

2}, Qingyu Guan^{1

2}, Chenyan Hong^{1

2}, Yufan Wei^{1

2}, Ningning Min^{1

2}, Aiying Qi², Yanjun Zhang², Xiru Li²

Affiliations

¹ School of Medicine, Nankai University, Tianjin, China.
² Department of General Surgery, Chinese People's Liberation Army General Hospital, Beijing, China.
³ Medical School of Chinese PLA, Beijing, China.

Abstract

Background: Triple-negative breast cancer (TNBC) is a malignant subtype of breast cancer, the main treatments for which are chemotherapy and surgery. PIK3CA is an oncogene that encodes the p110α subunit of class IA PI3K to regulate cell proliferation and apoptosis. Some reports have observed neoadjuvant chemotherapy (NAC) to have poor pathological complete response (pCR) rates in TNBC with PIK3CA mutation. This study aimed to explore the mechanism of how mutant PIK3CA alters chemotherapeutic susceptibility in TNBC.

Methods: TNBC cell lines (MDA-MB-231 and MDA-MB-468) with PIK3CA gene mutations (E545K and H1047R regions) and overexpression were established by transfection. NOD/SCID mice were used for in vivo experiments. Epirubicin was used as the chemotherapeutic agent. Cell viability, cell cycle, apoptosis, and Transwell assays were conducted for phenotype analysis. Western blot, quantitative reverse transcription-polymerase chain reaction, and immunohistochemistry were used to detect gene and protein expression levels. A clinical analysis of 50 patients with TNBC was also performed.

Results: Cell viability and Transwell assays showed that PIK3CA mutation promoted TNBC cell growth and conferred an enhanced migratory phenotype. Cell cycle and apoptosis assays showed that PIK3CA mutation moderately improved the proliferation ability of TNBC cells and remarkably inhibited their apoptosis. After epirubicin therapy, the proportion of early apoptotic cells decreased among cells with PIK3CA mutation. Further, xenograft tumors grew faster in NOD/SCID mice injected with mutated cell lines than in control group, suggesting that PIK3CA mutation caused chemotherapy resistance. Importantly, western blot and immunohistochemical analysis showed that cells and mouse tumors in the PIK3CA mutation groups exhibited different expression levels of apoptosis-related markers (Xiap, Bcl-2, and Caspase 3) and proteins associated with the PI3K/AKT/mTOR pathway (p110α, AKT, p-AKT, mTOR, p-mTOR, p-4E-BP1, p-p70S6K, and Pten). Moreover, prognostic analysis of 50 patients with TNBC indicated that PIK3CA mutation might be linked with relapse and death.

Conclusions: PIK3CA mutation confers resistance to chemotherapy in TNBC by inhibiting apoptosis and activating the PI3K/AKT/mTOR signaling pathway.

Keywords: PI3K/AKT/mTOR pathway; PIK3CA mutation; Triple-negative breast cancer (TNBC); apoptosis; chemotherapy resistance.