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. 2023 Oct;45(10):1329-1338.
doi: 10.1007/s13258-023-01437-y. Epub 2023 Aug 27.

Interference KRT17 reverses doxorubicin resistance in triple-negative breast cancer cells by Wnt/β-catenin signaling pathway

Liqiong Wu  1   2 , Wenshuang Ding  1   2 , Xiaopai Wang  1   2 , Xiubo Li  1   2 , Jing Yang  3   4
Affiliations

Interference KRT17 reverses doxorubicin resistance in triple-negative breast cancer cells by Wnt/β-catenin signaling pathway

Liqiong Wu et al. Genes Genomics. 2023 Oct.

Abstract

Background: Triple-negative breast cancer (TNBC) is a subtype of breast cancer with the highest degree of malignancy and is easily resistant to drugs due to the lack of hormone receptors. Research on the resistance mechanisms in TNBC is particularly important. Keratin 17 (KRT17) is highly expressed in TNBC. Anthracycline doxorubicin (Dox) is a commonly used chemotherapeutic drug for early stage triple-negative breast cancer.

Objective: This study investigated the role of KRT17 in TNBC-Dox resistance.

Methods: Immuno-histochemical staining, qPCR, western blotting (WB), and immunofluorescence were used to detect the expression of KRT17 in TNBC-Dox-resistant patients and in TNBC-Dox-resistant MDA-MB-468 and MDA-MB-231. the effect of KRT17 on the proliferation and migration in KRT17 knockdown of TNBC-Dox-resistant cells was determined by the CCK8, clone formation, transwell invasion and wound healing assays were used to determine.

Results: KRT17 was highly expressed in the TNBC-Dox-resistant cells. Knockdown of KRT17 significantly reduced the IC50s of TNBC-Dox-resistant and parental strains and also reduced the proliferation and invasion abilities of TNBC-Dox-resistant cell lines. KRT17 regulated the Wnt/β-catenin signaling pathway. The inhibitory effect of KRT17 knockdown on the proliferation and migration of TNBC-Dox-resistant cells was reversed by an activator of the Wnt signaling pathway.

Conclusion: KRT17 can inhibit the Wnt/β-catenin signaling pathway, thereby reducing the proliferation and invasion ability of TNBC-Dox-resistant cells.

Keywords: Doxorubicin resistance; KRT17; Triple-negative breast cancer; Wnt/β-catenin signaling pathway.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Fig. 1
Fig. 1
KRT17 is highly expressed in TNBC-Dox-resistant cells. (A) Constructed in MDA-MB-468-DOX and MDA-MB-231-DOX resistant cells respectively and tested for parental resistance to Dox resistant cells by assaying IC50. (B) qPCR detection of KRT17 expression in MDA-MB-468 and MDA-MB-231 parental and Dox-resistant cells. (C) KRT17 protein level detected by WB in MDA-MB-468 and MDA-MB-231 parental and Dox-resistant cells. (D) Immunofluorescence detection of KRT17 expression and cellular localization in MDA-MB-468 and MDA-MB-231 parental and Dox-resistant cells
Fig. 2
Fig. 2
KRT17 knockdown inhibits Dox resistance in TNBC cells. (A) qPCR detection of siRNA infection efficiency in MDA-MB-468-Dox and MDA-MB-231-Dox-resistant cells. (B, C) qPCR and WB detection of KRT17 knockdown in TNBC-Dox-resistant cells KRT17 stable cell line. (D) IC50s were detected in TNBC-Dox-resistant knockdown KRT17 stable cell lines
Fig. 3
Fig. 3
KRT17 knockdown inhibited the proliferation and migration of TNBC-Dox-resistant strains. (A) The effect of KRT17 knockdown on the proliferation of TNBC-Dox-resistant strains was detected by CCK8, with and without Dox. (B) The clone formation assay detected the monoclonal formation ability of KRT17 knockdown against TNBC-Dox-resistant strains, with and without Dox. (C) Transwell invasion assays used to detect the effect of knockdown of KRT17 on the invasion ability of TNBC-Dox-resistant strains, with and without Dox. (D) The effect of KRT17 knockdown on the growth and migration of TNBC-Dox-resistant strains was examined by the wound healing assay
Fig. 4
Fig. 4
KRT17 knockdown inhibits the Wnt/β-catenin signaling pathway. (A) WB detection of the effect of KRT17 knockdown on β-catenin expression in TNBC-Dox-resistant cells. (B) STRING database analysis of the regulatory network of KRT17 and Wnt/β-catenin signaling pathway. (C) The effect of KRT17 on the expression of APC and AXIN protein was detected by WB.
Fig. 5
Fig. 5
Wnt activator reverses the inhibitory effect of interfering KRT17 on proliferation and migration of TNBC-Dox-resistant strains. (A) The reversal effect of Wnt/β-catenin activator on the proliferation inhibition of TNBC-Dox-resistant strains by KRT17 knockdown was detected by CCK8. (B) Clone formation assay to detect the reversal effect of Wnt/β-catenin activator on the inhibition of monoclonal formation of TNBC-Dox-resistant strains by knockdown of KRT17. (C) Transwell invasion assays were used to detect the reversal effect of Wnt/β-catenin activator on KRT17 knockdown on the invasive ability of TNBC-Dox resistant strains. (D) Wound healing was used to detect the effect of Wnt/β-catenin activator on KRT17 knockdown on TNBC-Dox resistance reversal of inhibition of growth and migration ability of drug strains
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