Prognostic Relevance of HJURP Expression in Patients with Surgically Resected Colorectal Cancer
Abstract
:1. Introduction
2. Results
2.1. Association Between HJURP Expression and Cancer-Specific Deaths in CRC Patients
2.2. Transduction of CRC Cells by siRNA
2.3. Reduction of Colorectal Cancer Cell Proliferation by siRNA-Induced Downregulation of HJURP Expression
2.4. Suppression of Migration and Invasion of CRC Cells by Downregulation of HJURP
2.5. CRC Cell Anchorage-Independent Growth Reduction by HJURP
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Culture
4.2. Silencing of HJURP by siRNA
4.3. Reverse Transcriptase PCR
4.4. Western Blotting Assay
4.5. Cell Proliferation Assay
4.6. Cell Invasion and Migration Assay
4.7. Soft Agar Colony Formation Assay
4.8. Immunohistochemistry (IHC)
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
CRC | Colorectal cancer |
HJURP | Holliday Junction Recognition Protein |
CENP-A | Centromere Protein-A |
IHC | Immunohistochemistry |
siRNA | Small interfering RNA |
HR | Hazard ratio |
CI | Confidence Interval |
TNM | Tumor-Node-Metastasis |
CKI | Cyclin-dependent kinase inhibitor |
ERK | Extracellular Signal Regulated Kinase |
GSK3β | Glycogen Synthase Kinas 3 Beta |
MAPK | Mitogen-Activated Protein Kinase |
References
- Foltz, D.R.; Jansen, L.E.; Bailey, A.O.; Yates, J.R., III; Basset, E.A.; Wood, S.; Black, B.E.; Cleveland, D.W. Centromere-specific assembly of CENP-a nucleosomes is mediated by HJURP. Cell 2009, 137, 472–484. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dunleavy, E.M.; Roche, D.; Tagami, H.; Lacoste, N.; Ray-Gallet, D.; Nakamura, Y.; Daigo, Y.; Nakatani, Y.; Almouzni-Pettinotti, G. HJURP is a cell-cycle- dependent maintenance and deposition factor of CENP-A at centromeres. Cell 2009, 137, 485–497. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gurard-Levin, Z.A.; Quivy, J.P.; Almouzni, G. Histone chaperones: Assisting histone traffic and nucleosome dynamics. Annu. Rev. Biochem. 2014, 83, 487–517. [Google Scholar] [CrossRef]
- Kato, T.; Sato, N.; Hayama, S.; Yamabuki, T.; Ito, T.; Miyamoto, M.; Kondo, S.; Nakamura, Y.; Diago, Y. Activation of Holliday junction recognizing protein involved in the chromosomal stability and immortality of cancer cells. Cancer Res. 2007, 67, 8544–8553. [Google Scholar] [CrossRef] [Green Version]
- Montes de Oca, R.; Gurard-Levin, Z.A.; Berger, F.; Rehman, H.; Martel, E.; Corpet, A.; de Koning, L.; Vassias, I.; Wilson, L.O.W.; Meseure, D.; et al. The histone chaperone HJURP is a new independent prognostic marker for luminal A breast carcinoma. Mol. Oncol. 2015, 9, 657–674. [Google Scholar] [CrossRef]
- Jemal, A.; Bray, F.; Center, M.M.; Ferlay, J.; Ward, E.; Forman, D. Global cancer statistics. CA Cancer J. Clin. 2011, 61, 69–90. [Google Scholar] [CrossRef] [Green Version]
- Jemal, A.; Center, M.M.; DeSantis, C.; Ward, E.M. Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol. Biomark. Prev. 2010, 19, 1893–1907. [Google Scholar] [CrossRef] [Green Version]
- Van Cutsem, E.; Oliveira, J. Advanced colorectal cancer: ESMO clinical recommendations for diagnosis, treatment and follow-up. Ann. Oncol. 2009, 20, 61–63. [Google Scholar] [CrossRef]
- De Gramont, A.; Bosset, J.F.; Milan, C.; Rougier, P.; Bouche, O.; Morvan, P.L.E.; Louvet, C.; Guillot, T.; Francois, E.; Bedenne, L. Randomized trial comparing monthly low-dose leucovorin and fluorouracil bolus with bimonthly high-dose leucovorin and fluorouracil bolus plus continuous infusion for advanced colorectal cancer: A French intergroup study. J. Clin. Oncol. 1997, 15, 808–815. [Google Scholar] [CrossRef]
- Van Cutsem, E.; Hoff, P.M.; Harper, P.; Bukowski, R.M.; Cunningham, D.; Dufour, P.; Graeven, U.; Lokich, J.; Madajewicz, S.; Maroun, J.A.; et al. Oral capecitabine vs intravenous 5-fluorouracil and leucovorin: Integrated efficacy data and novel analyses from two large, randomised, phase III trials. Br. J. Cancer 2004, 90, 1190–1197. [Google Scholar] [CrossRef] [PubMed]
- Grothey, A.; Sargent, D.; Goldberg, R.M.; Schmoll, H.-J. Survival of patients with advanced colorectal cancer improves with the availability of fluorouracil-leucovorin, irinotecan, and oxaliplatin in the course of treatment. J. Clin. Oncol. 2004, 22, 1209–1214. [Google Scholar] [CrossRef] [PubMed]
- Tournigand, C.; André, T.; Achille, E.; Lledo, G.; Flesh, M.; Mery-Mignard, D.; Quinaux, E.; Couteau, C.; Buyse, M.; Ganem, G.; et al. FOLFIRI followed by FOLFOX6 or the reverse sequence in advanced colorectal cancer: A randomized GERCOR study. J. Clin. Oncol. 2004, 22, 229–237. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hurwitz, H.; Fehrenbacher, L.; Novotny, W.; Cartwright, T.; Hainsworth, J.; Heim, W.; Berlin, J.; Baron, A.; Griffing, S.; Holmgren, E.; et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N. Engl. J. Med. 2004, 350, 2335–2342. [Google Scholar] [CrossRef] [Green Version]
- Van Cutsem, E.; Köhne, C.H.; Láng, I.; Folprecht, G.; Nowacki, M.P.; Cascinu, S.; Shchepotin, I.; Maurel, J.; Cunningham, D.; Tejpar, S.; et al. Cetuximab plus irinotecan, fluorouracil, and leucovorin as first-line treatment for metastatic colorectal cancer: Updated analysis of overall survival according to tumor KRAS and BRAF mutation status. J. Clin. Oncol. 2011, 29, 2011–2019. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rawla, P.; Sunkara, T.; Barsouk, A. Epidemiology of colorectal cancer incidence, mortality, survival, and risk factors. Prz Gastroenterol. 2019, 14, 89–103. [Google Scholar] [CrossRef]
- Hu, B.; Wang, Q.; Wang, Y.; Chen, J.; Li, P.; Han, M. Holliday junction-recognizing protein promotes cell proliferation and correlates with unfavorable clinical outcome of hepatocellular carcinoma. Onco Targets Ther. 2017, 10, 2601–2607. [Google Scholar] [CrossRef] [Green Version]
- Chen, T.; Huang, H.; Zhou, Y.; Geng, L.; Shen, T.; Yin, S.; Zhou, L.; Zheng, S. Hepatocellular carcinoma proliferation by destabilizing p21 via the MAPK/ERK1/2 and AKT/GSK3β signaling pathways. J. Exp. Clin. Cancer Res. 2018, 37, 193. [Google Scholar] [CrossRef] [Green Version]
- Lacoste, N.; Woolfe, A.; Tachiwana, H.; Garea, A.V.; Barth, T.; Cataloube, S.; Kurumizaka, H.; Imhof, A.; Almouzni, G. Mislocalization of the centromeric histone variant CenH3/CENP-A in human cells depends on the chaperone DAXX. Mol Cell. 2014, 53, 631–644. [Google Scholar] [CrossRef] [Green Version]
- Tachiwana, H.; Muller, S.; Blumer, J.; Klare, K.; Mussacio, A.; Almouzni, G. HJURP involvement in de novo CenH3(CENP-A) and CENP-C recruitment. Cell Rep. 2015, 11, 22–32. [Google Scholar] [CrossRef]
- Hu, Z.; Huang, G.; Sadanandam, A.; Gu, S.; Lenburg, M.E.; Pai, M.; Bayani, N.; Blakely, E.A.; Gray, J.W.; Mao, J.-H. The expression level of HJURP has an independent prognostic impact and predicts the sensitivity to radiotherapy in breast cancer. Breast Cancer Res. 2010, 12, R18. [Google Scholar] [CrossRef] [Green Version]
- Johnston, K.; Joglekar, A.; Hori, T.; Suziki, A.; Fukagawa, T.; Salmon, E.D. Vertebrate kinetochore protein architecture: Protein copy number. J. Cell Biol. 2010, 176, 795–805. [Google Scholar] [CrossRef] [PubMed]
Clinicopathological Factors | HJURP Expression | Total (N = 162) | p Value | |
---|---|---|---|---|
Low (N = 118) | High (N = 44) | |||
Age, years, mean (SD) | 0.240 | |||
Gender, N (%) | 0.042 | |||
F | 41 (34.7) | 23 (52.4) | 64 (39.5) | |
M | 77 (65.3) | 21 (47.7) | 98 (60.5) | |
pT stage, N (%) | 0.541 | |||
1 and 2 | 20 916.9) | 7 (15.9) | 27 (16.7) | |
3 and 4 | 98 (83.1) | 37 (84.1) | 135 (83.3) | |
pN stage, N (%) | 0.797 | |||
negative | 59 (50.0) | 23 (52.3) | 82 (50.6) | |
positive | 59 (50.0) | 21 (47.7) | 80 (49.4) | |
Metastasis, N (%) | 0.729 | |||
negative | 114 (96.6) | 42 (95.5) | 156 (96.3) | |
positive | 4 (3.4) | 2 (4.5) | 6 (3.7) | |
Vascular invasion, N (%) | 0.891 | |||
negative | 95 (80.5) | 35 (79.5) | 130 (80.2) | |
positive | 23 (19.5) | 9 (20.5) | 32 (19.8) | |
Lymphatic invasion, N (%) | 0.677 | |||
negative | 87 (73.7) | 31 (70.5) | 118 (72.8) | |
positive | 31 (26.3) | 13 (29.5) | 44 (27.2) | |
* Stage, N (%) | 0.797 | |||
I and II | 59 (50.0) | 23 (52.3) | 82 (50.6) | |
III and IV | 59 (50.0) | 21 (47.7) | 80 (49.4) |
Clinicopathologic Factors | Variable | Univariate Analysis | Multivariate Analysis | ||
---|---|---|---|---|---|
Hazard Ratio (95%/CI) | p Value | Hazard Ratio (95%/CI) | p Value | ||
Age | <60 yr vs. ≥60 yr | 0.990 (0.607–11.615) | 0.967 | ||
Gender | Female vs. Male | 0.864 (9.549–1.360) | 0.528 | ||
Stage | I, II vs. III, IV | 1.974 (1.245–3.130) | 0.004 | 1.771 (1.087–2.886) | 0.022 |
Metastasis | Negative vs. Positive | 6.000 (2.522–14.272) | 0.000 | 3.559 (1.437–8.813) | 0.006 |
Vascular Invasion | Negative vs. Positive | 1.516 (0.893–2.575) | 0.124 | ||
Lymphatic Invasion | Negative vs. Positive | 2.026 (1.2264–3.247) | 0.003 | 1.573 (0.954–2.592) | 0.076 |
HJURP Expression | Low vs. High | 1.766 (1.103–2.829) | 0.018 | 1.880 (1.167–3.027) | 0.009 |
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Kang, D.H.; Woo, J.; Kim, H.; Kim, S.Y.; Ji, S.; Jaygal, G.; Ahn, T.S.; Kim, H.J.; Kwak, H.J.; Kim, C.-J.; et al. Prognostic Relevance of HJURP Expression in Patients with Surgically Resected Colorectal Cancer. Int. J. Mol. Sci. 2020, 21, 7928. https://doi.org/10.3390/ijms21217928
Kang DH, Woo J, Kim H, Kim SY, Ji S, Jaygal G, Ahn TS, Kim HJ, Kwak HJ, Kim C-J, et al. Prognostic Relevance of HJURP Expression in Patients with Surgically Resected Colorectal Cancer. International Journal of Molecular Sciences. 2020; 21(21):7928. https://doi.org/10.3390/ijms21217928
Chicago/Turabian StyleKang, Dong Hyun, Jongsoo Woo, Hyeongjoo Kim, Soo Youn Kim, Sanghee Ji, Gunn Jaygal, Tae Sung Ahn, Han Jo Kim, Hyoung Jong Kwak, Chang-Jin Kim, and et al. 2020. "Prognostic Relevance of HJURP Expression in Patients with Surgically Resected Colorectal Cancer" International Journal of Molecular Sciences 21, no. 21: 7928. https://doi.org/10.3390/ijms21217928