Identification of a 3,3-difluorinated tetrahydropyridinol compound as a novel antitumor agent for hepatocellular carcinoma acting via cell cycle arrest through disturbing CDK7-mediated phosphorylation of Cdc2
- 57 Downloads
Tetrahydropyridinol derivatives were recently reported to exhibit good biological activities, and the incorporation of fluorine into organic molecules may have profound effects on their physical and biological properties. Therefore, we investigated the anticancer activities of six fluorinated tetrahydropyridinol derivatives that we synthesized previously. We found that only one compound, 3,3-difluoro-2,2-dimethyl-1,6-diphenyl-5-tosyl-1,2,3,6-tetrahydropyridin-4-ol, showed significant antiproliferative activity on human hepatocellular carcinoma HepG2 and HMCCLM3 cells (the IC50 values were 21.25 and 29.07 μM, respectively). We also found that this compound mediated cell cycle arrest in the G0/G1 phase at 30–40 μM. Western blot analysis demonstrated that the cell cycle arrest induced by this compound in HepG2 and HMCCLM3 cells was associated with a significant decrease in Cdc2 and cyclin B1, which led to the accumulation of the phosphorylated-Tyr15 (inactive) form of Cdc2 and low expression of M phase-promoting factor (cyclin B1/Cdc2). Moreover, cells treated with this compound exhibited decreased expression of cyclin-dependent kinase (CDK)-activating kinase (CDK7/cyclin H). This compound also induced cell apoptosis via activation of caspase-3. A xenograft model in nude mice demonstrated anti-liver cancer activity and the mechanism of action of this compound. These findings indicated that the anticancer effect of this compound was partially due to G0/G1 cell cycle arrest via inhibition of CDK7-mediated expression of Cdc2, and this compound may be a promising anticancer candidate for further investigation.
KeywordsTetrahydropyridinol CDK7 Human hepatocellular carcinoma Cdc2
This work was supported by postdoctoral research station of Central South University and funded by the Hunan Provincial Natural Science Foundation of China (No. 2016JJ5013) and the National Natural Science Foundation of China (No. 81372516).
Compliance with ethical standards
The authors declare no competing interests.
This article does not contain any studies with human participants performed by any of the authors. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The Institute Research Ethics Committee of Central South University approved this experiment (No. 2018-SYDW-0111). All procedures performed in studies involving animals were in accordance with the ethical standards of the Animal Ethics Committee of Central South University(Changsha, Hunan Province, China) which the studies were conducted.
Informed consent is not required in this type of study.
- 2.Chen XY, Xiao EH, Shu DS, Yang CZ, Liang B, He Z, Bian DJ (2014) Evaluating the therapeutic effect of hepatocellular carcinoma treated with transcatheter arterial chemoembolization by magnetic resonance perfusion imaging. J Eur Gastroen Hepat 26:109–113. https://doi.org/10.1097/MEG.0b013e328363716e CrossRefGoogle Scholar
- 5.Aridoss G, Amirthaganesan S, Kumar NA, Kim JT, Lim KT, Kabilan S, Jeong YT (2008) A facile synthesis, antibacterial, and antitubercular studies of some piperidin-4-one and tetrahydropyridine derivatives. Bioorg Med Chem Lett 18:6542–6548. https://doi.org/10.1016/j.bmcl.2008.10.045 CrossRefGoogle Scholar
- 10.Leung TW, Tang AM, Zee B, Yu SC, Lai PB, Lau WY, Johnson PJ (2002) Factors predicting response and survival in 149 patients with unresectable hepatocellular carcinoma treated by combination cisplatin, interferon-alpha, doxorubicin and 5-fluorouracil chemotherapy. Cancer 94:421–427. https://doi.org/10.1002/cncr.10236 CrossRefGoogle Scholar
- 13.Tan M, Jing T, Lan KH, Neal CL, Li P, Lee S, Fang DX, Nagata YC, Liu JX, Arlinghaus R, Hung MC, Yu DH (2002) Phosphorylation on tyrosine-15 of p34 Cdc2 by ErbB2 inhibits p34 Cdc2 activation and is involved in resistance to taxol-induced apoptosis. Mol Cell 9:993–1004. https://doi.org/10.1016/s0959-8049(02)81195-9 CrossRefGoogle Scholar
- 15.Tassan JP, Jaquenoud M, Fry AM, Frutiger S, Hughes GJ, Nigg EA (1995) In vitro assembly of a functional human CDK7-cyclin H complex requires MAT1, a novel 36 kDa RING finger protein. EMBO J 14:5608–5617. https://doi.org/10.1002/j.1460-2075.1995.tb00248.x CrossRefGoogle Scholar
- 21.Yu J, Guo QL, You QD, Zhao L, Gu HY, Yang Y, Zhang HW, Tan Z, Wang XT (2007) Gambogic acid-induced G2/M phase cell-cycle arrest via disturbing CDK7-mediated phosphorylation of CDC2/p34 in human gastric carcinoma BGC-823 cells. Carcinogenesis 28:632–638. https://doi.org/10.1093/carcin/bgl168 CrossRefGoogle Scholar
- 22.Clark K, Ainscow E, Peall A, Thomson S, Leishman A, Elaine S, Ali S, Coombes R, Barrett A, Bah AK (2017) CT7001, a novel orally bio-available CDK7 inhibitor, is highly active in in-vitro and in-vivo models of AML. Blood 130:2645Google Scholar
- 23.Patel H, Periyasamy M, Sava GP, Bondke A, Slafer BW, Kroll SHB, Barbazanges M, Starkey R, Ottaviani S, Harrod A, Aboagye EO, Buluwela L, Fuchter MJ, Barrett AGM, Coombes RC, Ali S (2018) ICEC0942, an orally bioavailable selective inhibitor of CDK7 for cancer treatment. Mol Cancer Ther 17:1156–1166. https://doi.org/10.1158/1535-7163.MCT-16-0847 CrossRefGoogle Scholar
- 25.Zhao L, Chen Z, Wang J, Yang L, Zhao Q, Wang J, Qi Q, Mu R, You QD, Guo QL (2010) Synergistic effect of 5-fluorouracil and the flavanoid oroxylin a on HepG2 human hepatocellular carcinoma and on H22 transplanted mice. Cancer chemoth Pharm 65:481–489. https://doi.org/10.1007/s00280-009-1053-2 CrossRefGoogle Scholar
- 26.Mahavorasirikul W, Viyanant V, Chaijaroenkul W, Itharat A, Na-Bangchang K (2010) Cytotoxic activity of Thai medicinal plants against human cholangiocarcinoma, laryngeal and hepatocarcinoma cells in vitro. BMC Complem Altern M 10(55). https://doi.org/10.1186/1472-6882-10-55
- 28.Ma KL, He YH, Zhang HY, Fei Q, Niu DD, Wang DM, Ding X, Xu H, Chen XP, Zhu JD (2012) DNA methylation-regulated mir-193a-3p dictates resistance of hepatocellular carcinoma to 5-fluorouracil via repression of SRSF2 expression. J Biol Chem 287:5639–5649. https://doi.org/10.1074/jbc.M111.291229 CrossRefGoogle Scholar
- 30.Ohta K, Hoshino H, Wang JH, Ono S, Iida Y, Hata K, Huang SK, Colquhoun S, Hoon DS (2015) MicroRNA-93 activates c-met/PI3K/Akt pathway activity in hepatocellular carcinoma by directly inhibiting PTEN and CDKN1A. Oncotarget 6(3211). https://doi.org/10.18632/oncotarget.3085
- 32.Shi L, Wu LL, Chen ZG, Yang JR, Chen XF, Yu FY, Zheng F, Lin XY (2015) MiR-141 activates Nrf2-dependent antioxidant pathway via down-regulating the expression of Keap1 conferring the resistance of hepatocellular carcinoma cells to 5-fluorouracil. Cell Physiol Biochem 35:2333–2348. https://doi.org/10.1159/000374036 CrossRefGoogle Scholar