Abstract
Purpose
Maintaining telomeres by recruiting telomerase-to-chromosome ends is essential for cancer cell survival. Inhibiting telomerase recruitment to telomeres represents a novel strategy for telomere-based lung cancer therapy. However, approaches for interrupting telomerase recruitment for cancer therapy still need to be explored.
Methods
The telomere-binding protein TPP1 is responsible for recruiting telomerase to telomeres and synthesizing telomeres through the association between the oligosaccharide/oligonucleotide-binding (OB)-fold domain of TPP1 and telomerase reverse transcriptase. We overexpressed the TPP1 OB domain (TPP1-OB) by lentivirus infection in lung cancer cells. Telomere length was examined by Southern blot analysis of terminal restriction fragments. The effects of TPP1-OB on cell proliferation, the cell cycle, apoptosis, chemosensitivity, and tumor growth were evaluated in vitro and in vivo.
Result
TPP1-OB inhibited the recruitment of telomerase to telomeres and shortened telomere length by acting as a dominant-negative mutant of TPP1. TPP1-OB resulted in reduced cell proliferation, G1 cell cycle arrest, and increased cell apoptosis in lung cancer cells. Cell apoptosis occurred mainly through the caspase-3-dependent signaling pathway. TPP1-OB also suppressed anchorage-independent growth and tumor growth in vivo. Moreover, we demonstrated that TPP1-OB enhances the sensitivity of lung cancer cells to the chemotherapeutic drug paclitaxel.
Conclusion
Our results suggest that inhibiting TPP1-mediated telomerase recruitment by expressing the TPP1-OB domain is a potential novel strategy for telomere-targeted lung cancer therapy.
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Abbreviations
- OB:
-
Oligosaccharide/oligonucleotide-binding
- TERT:
-
Telomerase reverse transcriptase
- TEL:
-
TPP1 glutamate (E) and leucine (L)-rich
- TEN:
-
Telomerase essential N-terminal
- TIF:
-
Telomere dysfunction-induced foci
- FBS:
-
Fetal bovine serum
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- FISH:
-
Fluorescence in situ hybridization
- PNA:
-
Peptide nucleic acids
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Funding
This work was supported by the National Natural Science Foundation of China (81472182 to BZ), the Young Innovative Talent Program of Tianjin Medical University Cancer Institute and Hospital (to BZ), and the Tianjin Thousand Talents program (to GG).
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Zhu, J., Liu, W., Chen, C. et al. TPP1 OB-fold domain protein suppresses cell proliferation and induces cell apoptosis by inhibiting telomerase recruitment to telomeres in human lung cancer cells. J Cancer Res Clin Oncol 145, 1509–1519 (2019). https://doi.org/10.1007/s00432-019-02921-3
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DOI: https://doi.org/10.1007/s00432-019-02921-3