As an inhibitor of heat shock proteins (HSPs), KNK437 has been reported to play an anti-tumor role in several cancers. But its therapeutic effect and mechanisms in colorectal cancer (CRC) remain unclear. Here, KNK437 sharply inhibited the level of DnaJ heat shock protein family (Hsp40) member A1 (DNAJA1), followed by DNAJB1, but had little effect on the levels of HSP27, HSP105, HSP90, and HSP70 in CRC cells. DNAJA1 promoted CRC cell proliferation in vitro and tumor growth and metastasis in vivo. Mechanistically, DNAJA1 was activated by E2F transcription factor 1 (E2F1) and then promoted cell cycle by stabilizing cell division cycle protein 45 (CDC45), which could be reversed by KNK437. DNAJA1 was significantly upregulated in CRC tissues and positively correlated with serosa invasion, lymph node metastasis. High level of DNAJA1 predicted poor prognosis for CRC patients. Its expression was highly linked with E2F1 and CDC45 in CRC tissues. More importantly, KNK437 significantly suppressed the growth of DNAJA1 expressing tumor in vivo. The combined treatment of KNK437 with 5-FU/L-OHP chemotherapy reduced liver metastasis of CRC. These data reveal a novel mechanism of KNK437 in anti-tumor therapy of CRC and provides a newly therapeutic strategy with potential translation to the CRC patients.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $51.94 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin. 2016;66:7–30.
Jaattela M. Heat shock proteins as cellular lifeguards. Ann Med. 1999;31:261–71.
Matijasevic Z, Snyder JE, Ludlum DB. Hypothermia causes a reversible, p53-mediated cell cycle arrest in cultured fibroblasts. Oncol Res. 1998;10:605–10.
Sonna LA, Fujita J, Gaffin SL, Lilly CM. Effects of heat and cold stress on mammalian gene expression. J Appl Physiol. 2002;92:1725–42.
Haggerty TJ, Dunn IS, Rose LB, Newton EE, Pandolfi F, Kurnick JT. Heat shock protein-90 inhibitors enhance antigen expression on melanomas and increase T cell recognition of tumor cells. PLoS ONE. 2014;12:e114506.
Bodoor K, Jalboush SA, Matalka I, Abu-Sheikha A, Waq RA, Ebwaini H, et al. Heat shock protein association with clinico-pathological characteristics of gastric cancer in Jordan: HSP70 is predictive of poor prognosis. Asian Pac J Cancer Prev. 2016;17:3929–37.
Thorsteinsdottir J, Stangl S, Fu P, Guo K, Albrecht V, Eigenbrod S, et al. Overexpression of cytosolic, plasma membrane bound and extracellular heat shock protein 70 (Hsp70) in primary glioblastomas. J Neurooncol. 2017;135:443–52.
Jagadish N, Agarwal S, Gupta N, Fatima R, Devi S, Kumar V, et al. Heat shock protein 70-2 (HSP70-2) overexpression in breast cancer. J Exp Clin Cancer Res. 2016;35:150.
Jhaveri K, Wang R, Teplinsky E, Chandarlapaty S, Solit D, Cadoo K, et al. A phase I trial of ganetespib in combination with paclitaxel and trastuzumab in patients with human epidermal growth factor receptor-2 (HER2)-positive metastatic breast cancer. Breast Cancer Res. 2017;19:89.
Wang Y, Liu H, Diao L, Potter A, Zhang J, Qiao Y, et al. Hsp90 inhibitor ganetespib sensitizes non-small cell lung cancer to radiation but has variable effects with chemoradiation. Clin Cancer Res. 2016;22:5876–86.
Lee HG, Park WJ, Shin SJ, Kwon SH, Cha SD, Seo YH, et al. Hsp90 inhibitor SY-016 induces G2/M arrest and apoptosis in paclitaxel-resistant human ovarian cancer cells. Oncol Lett. 2017;13:2817–22.
Ramalingam S, Goss G, Rosell R, Schmid-Bindert G, Zaric B, Andric Z, et al. A randomized phase II study of ganetespib, a heat shock protein 90 inhibitor, in combination with docetaxel in second-line therapy of advanced non-small cell lung cancer (GALAXY-1). Ann Oncol. 2015;26:1741–8.
Ban HS, Naik R, Kim HM, Kim BK, Lee H, Kim I, et al. Identification of targets of the HIF-1 Inhibitor IDF-11774 using alkyne-conjugated photoaffinity probes. Bioconjugugate Chem. 2016;27:1911–20.
Taba K, Kuramitsu Y, Ryozawa S, Yoshida K, Tanaka T, Mori-Iwamoto S, et al. KNK437 downregulates heat shock protein 27 of pancreatic cancer cells and enhances the cytotoxic effect of gemcitabine. Chemotherapy. 2011;57:12–6.
Sahin E, Sahin M, Sanlioğlu AD, Gümüslü S. KNK437, a benzylidene lactam compound, sensitises prostate cancer cells to the apoptotic effect of hyperthermia. Int J Hyperth. 2011;27:63–73.
Yokota S, Kitahara M, Nagata K. Benzylidene lactam compound, KNK437, a novel inhibitor of acquisition of thermotolerance and heat shock protein induction in human colon carcinoma cells. Cancer Res. 2000;60:2942–8.
Terada K, Yomogida K, Imai T, Kiyonari H, Takeda N, Kadomatsu T, et al. A type I DnaJ homolog, DjA1, regulates androgen receptor signaling and spermatogenesis. EMBO J. 2005;24:611–22.
Wang CC, Liao YP, Mischel PS, Iwamoto KS, Cacalano NA, McBride WH. HDJ-2 as a target for radiosensitization of glioblastoma multiforme cells by the farnesyltransferase inhibitor R115777 and the role of the p53/p21 pathway. Cancer Res. 2006;66:6756–62.
Stark JL, Mehla K, Chaika N, Acton TB, Xiao R, Singh PK, et al. Structure and function of human DnaJ homologue subfamily a member 1 (DNAJA1) and its relationship to pancreatic cancer. Biochemistry. 2014;53:1360–72.
Banerji U, O’Donnell A, Scurr M, Pacey S, Stapleton S, Asad Y, et al. Phase I pharmacokinetic and pharmacodynamic study of 17-allylamino, 17-demethoxygel- danamycin in patients with advanced malignancies. J Clin Oncol. 2005;23:4152–61.
Hadchity E, Aloy MT, Paulin C, Armandy E, Watkin E, Rousson R, et al. Heat shock protein 27 as a new therapeutic target for radiation sensitization of head and neck squamous cell carcinoma. Mol Ther. 2009;17:1387–94.
Kudryavtsev VA, Khokhlova AV, Mosina VA, Selivanova EI, Kabakov AE. Induction of Hsp70 in tumor cells treated with inhibitors of the Hsp90 activity: a predictive marker and promising target for radiosensitization. PLoS ONE. 2017;12:e0173640.
Kimura A, Ogata K, Altan B, Yokobori T, Mochiki E, Yanai M, et al. Nuclear heat shock protein 110 expression is associated with poor prognosis and hyperthermo-chemotherapy resistance in gastric cancer patients with peritoneal metastasis. World J Gastroenterol. 2017;23:7541–50.
Kuramitsu Y, Wang Y, Taba K, Suenaga S, Ryozawa S, Kaino S, et al. Heat-shock protein 27 plays the key role in gemcitabine-resistance of pancreatic cancer cells. Anticancer Res. 2012;32:2295–9.
Oommen D, Prise KM. KNK437, abrogates hypoxia-induced radioresistance by dual targeting of the AKT and HIF-1α survival pathways. Biochem Biophys Res Commun. 2012;421:538–43.
Barrio S, Gallardo M, Arenas A, Ayala R, Rapado I, Rueda D, et al. Inhibition of related JAK/STAT pathways with molecular targeted drugs shows strong synergy with ruxolitinib in chronic myeloproliferative neoplasm. Br J Haematol. 2013;161:667–76.
Qiu XB, Shao YM, Miao S, Wang L. The diversity of the DnaJ/Hsp40 family, the crucial partners for Hsp70 chaperones. Cell Mol Life Sci. 2006;63:2560–70.
Walsh P, Bursać D, Law YC, Cyr D, Lithgow T. The J-protein family: modulating protein assembly, disassembly and translocation. EMBO Rep. 2004;5:567–71.
Kotlarz A, Tukaj S, Krzewski K, Brycka E, Lipinska B. Human Hsp40 proteins, DNAJA1 and DNAJA2, as potential targets of the immune response triggered by bacterial DnaJ in rheumatoid arthritis. Cell Stress Chaperon-. 2013;18:653–9.
Gibbs SJ, Braun JE. Emerging roles of J proteins in neurodegenerative disorders. Neurobiol Dis. 2008;32:196–9.
Meshalkina DA, Shevtsov MA, Dobrodumov AV, Komarova EY, Voronkina IV, Lazarev VF, et al. Knock-down of Hdj2/DNAJA1 co-chaperone results in an unexpected burst of tumorigenicity of C6 glioblastoma cells. Oncotarget. 2016;7:22050–63.
Chen HZ, Tsai SY, Leone G. Emerging roles of E2Fs in cancer: an exit from cell cycle control. Nat Rev Cancer. 2009;9:785–97.
Tanaka H, Matsumura I, Ezoe S, Satoh Y, Sakamaki T, Albanese C, et al. E2F1 and c-Myc potentiate apoptosis through inhibition of NF-kappaB activity that facilitates MnSOD-mediated ROS elimination. Mol Cell. 2002;9:1017–29.
Bracken AP, Ciro M, Cocito A, Helin K. E2F target genes: unraveling the biology. Trends Biochem. 2004;29:409–17.
Ohtani K, DeGregori J, Nevins JR. Regulation of the cyclin E gene by transcription factor E2F1. Proc Natl Acad Sci USA. 1995;92:12146–50.
Bauerschmidt C, Pollok S, Kremmer E, Nasheuer HP, Grosse F. Interactions of human Cdc45 with the Mcm2-7 complex, the GINS complex, and DNA polymerases delta and epsilon during S phase. Genes Cells. 2007;12:745–58.
Hardy CF. Identification of Cdc45p, an essential factor required for DNA replication. Gene. 1997;187:239–46.
Nasheuer HP, Smith R, Bauerschmidt C, Grosse F, Weisshart K. Initiation of eukaryotic DNA replication: regulation and mechanisms. Prog Nucleic Acid Res Mol Biol. 2002;72:41–94.
Sun J, Shi R, Zhao S, Li X, Lu S, Bu H, et al. Cell division cycle 45 promotes papillary thyroid cancer progression via regulating cell cycle. Tumour Biol. 2017;39:1010428317705342.
Li HY, Jin N, Han YP, Jin XF. Pathway crosstalk analysis in prostate cancer based on protein-protein network data. Neoplasma. 2017;64:22–31.
Tane S, Sakai Y, Hokka D, Okuma H, Ogawa H, Tanaka Y, et al. Significant role of Psf3 expression in non-small-cell lung cancer. Cancer Sci. 2015;106:1625–34.
Haider S, Jun W, Nagano A, Desai A, Arumugam P, Dumartin L, et al. A multi-gene signature predicts outcome in patients with pancreatic ductal adenocarcinoma. Genome Med. 2014;6:105.
Parrales A, Ranjan A, Iyer SV, Padhye S, Weir SJ. DNAJA1 controls the fate of misfoldedmutant p53 through the mevalonate pathway. Nat Cell Biol. 2016;18:1233–43.
This work was supported by the National key R&D program of China (2017YFC1309002), National Basic Research Program of China (973 Program, 2015CB554002), National Natural Science Foundation of China (81672821, 81472313, 81773101, 81272759, 81401927, 81302151, and 81802306), Project funded by China Postdoctoral Science Foundation (2018M633081 and 2018M633079), Natural Science Foundation of Guangdong Province (2018A030310457).
Conflict of interest
The authors declare that they have no conflict of interest.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.