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Title[redir]:
Targeting DTL induces cell cycle arrest and senescence and suppresses | OTT
Description:
Targeting DTL induces cell cycle arrest and senescence and suppresses cell growth and colony formation through TPX2 inhibition in human hepatocellular carcinoma cells Yu-Chia Chen,1,* I-shu Chen,1,* Guan-Jin Huang,2 Chi-hsiang Kang,1 Kuo-Chiang Wang,1 Min-Jen Tsao,3 Hung-Wei Pan4,5 1Department of Surgery, Division of General Surgery, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan; 2Department of Pathology, National Chung Kung University Hospital, Tainan, Taiwan; 3Department of General Surgery, Zuoying Branch of Kaohsiung Armed Forces General Hospital, Kaohsiung, Taiwan; 4Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan; 5Department of Applied Chemistry, National Pingtung University, Pingtung, Taiwan *These authors contributed equally to this work Background: Hepatocellular carcinoma (HCC) has an increasing incidence and high mortality. Surgical operation is not a comprehensive strategy for liver cancer. Moreover, tolerating systemic chemotherapy is difficult for patients with HCC because hepatic function is often impaired due to underlying cirrhosis. Therefore, a comprehensive strategy for cancer treatment should be developed. DTL (Cdc10-dependent transcript 2) is a critical regulator of cell cycle progression and genomic stability. In our previous study, the upregulation of DTL expression in aggressive HCC correlated positively with tumor grade and poor patient survival. We hypothesize that targeting DTL may provide a novel therapeutic strategy for liver cancer. DTL small interference RNAs were used to knock down DTL protein expression.Methods: A clonogenic assay, immunostaining, double thymidine block, imaging flow cytometry analysis, and a tumor spheroid formation assay were used to analyze the role of DTL in tumor cell growth, cell cycle progression, micronucleation, ploidy, and tumorigenicity.Results: Our results demonstrated that targeting DTL reduced cell cycle regulators and chromosome segregation genes, resulting in increased cell micronucleation. DTL depletion inhibited liver cancer cell growth, increased senescence, and reduced tumorigenesis. DTL depletion resulted in the disruption of the mitotic proteins cyclin B, CDK1, securin, seprase, Aurora A, and Aurora B as well as the upregulation of the cell cycle arrest gene p21. A rescue assay indicated that DTL should be targeted through TPX2 downregulation for cancer cell growth inhibition. Moreover, DTL silencing inhibited the growth of patient-derived primary cultured HCC cells.Conclusion: Our study results indicate that DTL is a potential novel target gene for treating liver cancer through liver cancer cell senescence induction. Furthermore, our results provide insights into molecular mechanisms for targeting DTL in liver cancer cells. The results also indicate several other starting points for future preclinical and clinical studies on liver cancer treatment. Keywords: DTL, cell cycle, senescence, TPX2, hepatocellular carcinoma
Title[redir]:
Targeting DTL induces cell cycle arrest and senescence and suppresses | OTT
Description:
Targeting DTL induces cell cycle arrest and senescence and suppresses cell growth and colony formation through TPX2 inhibition in human hepatocellular carcinoma cells Yu-Chia Chen,1,* I-shu Chen,1,* Guan-Jin Huang,2 Chi-hsiang Kang,1 Kuo-Chiang Wang,1 Min-Jen Tsao,3 Hung-Wei Pan4,5 1Department of Surgery, Division of General Surgery, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan; 2Department of Pathology, National Chung Kung University Hospital, Tainan, Taiwan; 3Department of General Surgery, Zuoying Branch of Kaohsiung Armed Forces General Hospital, Kaohsiung, Taiwan; 4Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan; 5Department of Applied Chemistry, National Pingtung University, Pingtung, Taiwan *These authors contributed equally to this work Background: Hepatocellular carcinoma (HCC) has an increasing incidence and high mortality. Surgical operation is not a comprehensive strategy for liver cancer. Moreover, tolerating systemic chemotherapy is difficult for patients with HCC because hepatic function is often impaired due to underlying cirrhosis. Therefore, a comprehensive strategy for cancer treatment should be developed. DTL (Cdc10-dependent transcript 2) is a critical regulator of cell cycle progression and genomic stability. In our previous study, the upregulation of DTL expression in aggressive HCC correlated positively with tumor grade and poor patient survival. We hypothesize that targeting DTL may provide a novel therapeutic strategy for liver cancer. DTL small interference RNAs were used to knock down DTL protein expression.Methods: A clonogenic assay, immunostaining, double thymidine block, imaging flow cytometry analysis, and a tumor spheroid formation assay were used to analyze the role of DTL in tumor cell growth, cell cycle progression, micronucleation, ploidy, and tumorigenicity.Results: Our results demonstrated that targeting DTL reduced cell cycle regulators and chromosome segregation genes, resulting in increased cell micronucleation. DTL depletion inhibited liver cancer cell growth, increased senescence, and reduced tumorigenesis. DTL depletion resulted in the disruption of the mitotic proteins cyclin B, CDK1, securin, seprase, Aurora A, and Aurora B as well as the upregulation of the cell cycle arrest gene p21. A rescue assay indicated that DTL should be targeted through TPX2 downregulation for cancer cell growth inhibition. Moreover, DTL silencing inhibited the growth of patient-derived primary cultured HCC cells.Conclusion: Our study results indicate that DTL is a potential novel target gene for treating liver cancer through liver cancer cell senescence induction. Furthermore, our results provide insights into molecular mechanisms for targeting DTL in liver cancer cells. The results also indicate several other starting points for future preclinical and clinical studies on liver cancer treatment. Keywords: DTL, cell cycle, senescence, TPX2, hepatocellular carcinoma
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Keywords {🔍}
cells, cell, dtl, cancer, cycle, tpx, protein, liver, growth, expression, figure, sirna, hcc, results, skhep, dna, depletion, rtpcr, assay, senescence, carcinoma, phase, targeting, hepatocellular, progression, arrest, mrna, measured, gene, inhibition, study, human, cyclin, levels, tumor, analysis, increased, control, medium, quantitative, egfptpx, aurora, target, damage, transfected, kaohsiung, treatment, rna, data, senescent,
Topics {✒️}
sirna-treated sk-hep-1 cells vectors expressing egfp-h2b egfp-tpx2 fusion protein dtl-depleted sk-hep-1 cells real-time rt-pcr assays denticleless/ra-regulated nuclear matrix quantitative real-time rt-pcr citric acid/sodium phosphate control rnai-oligonucleotide-transfected cells e2f1-dependent gene transcription sk-hep-1 cell line β-gal staining solution si-dtl egfp-tpx-2 cells micronucleated sk-hep-1 cells egfp-h2b cell line anchorage-independent cell growth cell lines derived microscopic low-power field human beta-actin gene dove medical press targeting pr-set7/set8 sa-β-gal assay including fast-track processing sk-hep-1 cells transfected real-time rt-pcr plated sk-hep-1 cells cell cycle-regulated nuclear p53-transactivated proapoptotic genes cul4-based e3 ligases completely contact inhibited endpoint rt-pcr assays egfp-tpx2 protein levels endpoint rt-pcr assay expression vector pegfp-c1 multiple cell-cycle regulator stage-independent prognostic factor egfp-tpx2 cells compared treating advanced hcc induces cell apoptosis35 dr wen-huei chang quantitative rt-pcr assay target/β-actin ratio manuscript login prof hey-chi hsu hepatocyte growth factor favored authors cdna microarray analysis γ-tubulin protein expression sk-hep-1 cells contact-inhibited cells
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