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Silencing GS Homeobox 2 Alleviates Gemcitabine Resistance in Pancreatic Cancer Cells by Activating SHH/GLI1 Signaling Pathway

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Abstract

Sonic hedgehog (SHH) signaling pathway and glioma-associated oncogene homolog 1 (GLI1) play important roles in the initiation and progression of pancreatic ductal adenocarcinoma (PDAC). GS homeobox 2 (GSX2, formerly GSH2) is a downstream target of SHH signaling, but its role in pancreatic cancer remains unclear. This study evaluates the role of GSH2 in the development and drug resistance of pancreatic cancer. Both cell culture and xenograft mouse model were used. Immunohistochemistry, Western blotting and quantitative RT-PCR were used to examine the expression of GSH2 and other related molecules. CCK8 assay was used to test the cell proliferation, and flow cytometry used to examine cell apoptosis upon gemcitabine treatment. It was found that GSH2 is overexpressed in human pancreatic cancer tissues and cells. The expression of SHH and GLI1 was reversely correlated with GSH2 in pancreatic cancer cells. SHH and GLI1 have protein–protein interactions with GSH2. GSH2 silencing in pancreatic cancer cells inhibited cell proliferation, migration and invasion, increased cell apoptosis and sensitized pancreatic cancer cells to gemcitabine treatment. Furthermore, in vivo study demonstrated that silencing GSH2 increased the efficacy of gemcitabine-based treatment. Our results indicate that GSH2 is overexpressed in pancreatic cancer. GSH2 silencing in pancreatic cancer alleviates gemcitabine resistance by activating SHH/GLI1 pathway. Thus, targeting GSH2 in PDAC could be a novel cancer therapeutic strategy.

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Abbreviations

GSX2, Formerly GSH2:

GS homeobox 2

SHH:

Sonic hedgehog

HH:

Hedgehog

PDAC:

Pancreatic ductal adenocarcinoma

GEM:

Gemcitabine

qRT-PCR:

Quantitative real-time PCR

GLI1:

Glioma-associated oncogene homolog 1

IHC:

Immunohistochemistry

SMO:

Smoothened

siRNA:

Small interfering RNA

si-Con:

Transfected with negative control siRNA

si-GSH2:

Transfected with GSH2 siRNA

MGE:

Medial ganglionic eminence

NKX2.1:

NK2 Homeobox 1

HPDE:

Human pancreatic duct epithelial

Co-IP:

Co-immunoprecipitation

ECM:

Extracellular matrix

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Author notes

  1. Lu Zhuang, Yao Yao and Lisi Peng have contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, Changhai Hospital, Navy Military Medical University, 168 Changhai Road, Shanghai, 200433, China

    Lu Zhuang, Yao Yao, Lisi Peng, Fang Cui, Cui Chen, Yang Zhang, Liqi Sun, Qihong Yu & Kun Lin

  2. Shanghai Hongkou District Jiaxing Road Subdistrict Community Healthcare Service Center, 1 Hongguan Road, Shanghai, 200086, China

    Lu Zhuang

  3. Department of Critical Care Medicine, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200030, China

    Yao Yao

  4. Department of Gastroenterology, Zhongnan Hospital, Wuhan University, 169 Donghu Road, Wuhan, 430071, China

    Kun Lin

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  1. Lu Zhuang
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Correspondence to Kun Lin.

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All procedures performed in this study were approved by the Changhai Hospital Institutional Review Board.

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Zhuang, L., Yao, Y., Peng, L. et al. Silencing GS Homeobox 2 Alleviates Gemcitabine Resistance in Pancreatic Cancer Cells by Activating SHH/GLI1 Signaling Pathway. Dig Dis Sci 67, 3773–3782 (2022). https://doi.org/10.1007/s10620-021-07262-1

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