American Association for Cancer Research
Browse

Supplementary Figure S1 from PTK6 Potentiates Gemcitabine-Induced Apoptosis by Prolonging S-phase and Enhancing DNA Damage in Pancreatic Cancer

Download (229.67 kB)
figure
posted on 2023-04-03, 17:09 authored by Hiroaki Ono, Marc D. Basson, Hiromichi Ito

Supplementary Figure S1. (A) The PTK6 gene silencing by alternative siRNA targeting different sequences of PTK6. (B) Effect of PTK6 gene-silencing on cytotoxity of gemcitabine. Cancer cells were transfected either with siRNA targeting to PTK6 or non-specific control siRNA for 48 hours prior to treatment with gemcitabine. Then cells were treated with gemcitabine at 25nM for MIAPaCa2 and 1 μM for Panc1 for 96 hrs.

History

ARTICLE ABSTRACT

Protein Tyrosine Kinase 6 (PTK6) is a non-receptor–type tyrosine kinase known to be expressed in various cancers, including pancreatic cancer. The role of PTK6 in cancer chemoresistance remains unclear. Therefore, it was hypothesized that PTK6 mechanistically regulates gemcitabine resistance in pancreatic cancer. Gemcitabine treatment stimulated endogenous PTK6 overexpression in MIAPaCa2 and Panc1 cells. PTK6 gene silencing increased cell survival after gemcitabine treatment and decreased apoptosis, whereas PTK6 overexpression decreased cell survival and increased apoptosis. Selection for gemcitabine resistance revealed substantially lower PTK6 expression in the gemcitabine-resistant subclones compared with the parental lines, while restoring PTK6 rescued gemcitabine sensitivity. Gemcitabine induced phosphorylation of H2AX (γ-H2AX) and ataxia-telangiectasia mutated kinase (pATM), specific markers for DNA double-strand breaks. Both gemcitabine-induced phosphorylation of H2AX and ATM were reduced by PTK6 knockdown and increased by PTK6 overexpression. PTK6 overexpression also increased the S-phase fraction 48 hours after gemcitabine treatment. Although gemcitabine activated both caspase-8 (CASP8) and caspase-9 (CASP9), the effect of PTK6 on gemcitabine-induced apoptosis required CASP8 but not CASP9. In mouse xenografts, PTK6 overexpression in subcutaneous tumors attenuated tumor growth after gemcitabine treatment. In conclusion, PTK6 prolongs S-phase and increases the ability of gemcitabine to cause DNA damage in vitro and in vivo.Implications: PTK6 affects cell cycle and DNA damage, thus making it an important therapeutic target to improve the outcomes of patients with pancreatic cancer. Mol Cancer Res; 13(8); 1174–84. ©2015 AACR.

Usage metrics

    Molecular Cancer Research

    Licence

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC