Mechanism of Calpain-2 resistance to paclitaxel through induction of epithelial mesenchymal transition in triple-negative breast cancer cells

doi:10.11958/20230208

Abstract

Abstract:

Objective To investigate the role and mechanism of Calpain-2 in paclitaxel (PTX) resistance in triple-negative breast cancer cells. Methods Human triple negative breast cancer MDA-MB-231 cells were transfected with Calpain-2 plasmid and corresponding empty vector, and cells were used as the overexpression group and the empty vector group. The untransfected cells were used as the blank group. MTT assay was performed to detect the effect of PTX on the survival rate of each group of cells. Western blot assay was performed to detect the expression of Calpain-2, E-calmodulin (E-cadherin), N-cadherin, Vimentin, phosphorylated large tumour suppressor gene 1 (p-LATS1) and phosphorylated yes-associated protein (p-YAP). Immunofluorescence assay was performed to detect the distribution of YAP. YAP inhibitor XMU-MP-1 was added to intervene on the basis of overexpression of Calpain-2. The mediated effect of YAP on PTX resistance was observed. Results Compared with the empty vector group, the survival rate of cells increased after PTX treatment in the overexpression group, and the IC₅₀ value increased (P<0.05). Compared with the empty vector group, N-cadherin and Vimentin protein expression were up-regulated, E-cadherin protein expression was down-regulated in the overexpression group (P<0.05), and p-LATS1 and p-YAP protein expression were up-regulated (P<0.01). YAP distribution in nucleus was reduced in the overexpression group. Compared with the overexpression group, N-cadherin and Vimentin protein expression were down-regulated and E-cadherin protein expression was up-regulated in the XMU-MP-1+ overexpression group (P<0.01). After PTX treatment, cell survival was reduced, and IC₅₀ was lower in the XMU-MP-1+ overexpression group compared with the overexpression group (P<0.01). Conclusion Calpain-2 induces epithelial mesenchymal transition in triple-negative breast cancer cells via YAP, resulting in drug resistance to PTX.

Key words: triple negative breast neoplasms, epithelial-mesenchymal transition, drug resistance, neoplasm, taxol, calpain 2

CLC Number:

R737.9

Figures/Tables 6

Fig.1 Role of Calpain-2 in the resistance of triple-negative breast cancer cells to PTX

Fig.2 Effect of Calpain-2 on EMT of triple-negative breast cancer cells

Fig.3 Effect of Calpain-2 on the expression of YAP

Fig.4 Effect of Calpain-2 on the distribution of YAP (immunofluorescence, ×400)

Fig.5 Role of YAP inhibition in Calpain-2 induced EMT in triple negative breast cancer cells

Fig.6 Inhibition of YAP in Calpain-2-induced PTX resistance in triple-negative breast cancer cells

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