Sains Malaysiana 50(7)(2021): 1935-1946

http://doi.org/10.17576/jsm-2021-5007-09

A Preliminary Study on the Impact of UCK2 Knockdown in DLD-1 Colorectal Cells Treated with DHODH Inhibitor

(Kajian Awal ke atas Kesan Penyahfungsian UCK2 di dalam Sel Kolorektum DLD-1 yang Dirawat dengan Perencat DHODH)

MOHAMAD FAIRUS ABDUL KADIR^1,2, PUTERI SHAFINAZ ABDUL-RAHMAN¹, KAVITHA NELLORE³ & SHATRAH OTHMAN¹*

¹Aurigene Discovery Technologies (M) Sdn. Bhd., Level 2, Research Management and Innovation Complex, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

²Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

³Aurigene Discovery Technologies Limited, 39-40, KIADB Industrial Area, Electronic City Phase II, Hosur Road, Bangalore 560100 Karnataka, India

Received: 22 October 2019/Accepted: 17 November 2020

ABSTRACT

Brequinar sodium (BQR) is a well-studied inhibitor of the dihydroorotate dehydrogenase (DHODH) enzyme. Both the DHODH and uridine-cytidine kinase 2 (UCK2) enzymes have been reported to be over-expressed in cancer cells to maintain the cells high demand for DNA and RNA for their proliferation. In this study, we aim to further sensitize cells to the effects of BQR by knocking down the UCK2 activity. In DLD-1 UCK2 knockdown cells, no change in the sensitivity of cells to BQR was observed. Uridine is known to reverse the anti-proliferative effect of DHODH inhibitors via the salvage pathway. We observed abrogation of approximately 30% of the uridine reversal effect in UCK2 knockdown cells compared to the wild type cells. Our finding indicates that the loss of UCK2 activity in the salvage pathway did not enhance the BQR-mediated cell proliferation inhibition but it abrogates the uridine reversal in the cells.

Keywords: BQR; DHODH; TAS-106; UCK2; uridine abrogation

ABSTRAK

Natrium Brequinar (BQR) dikenali sebagai salah satu perencat enzim dihidroorotat dehidrogenase (DHODH). Kedua-dua enzim DHODH dan uridina-sitidina kinase 2 (UCK2) diekspreskan secara berlebihan di dalam sel kanser untuk mengekalkan permintaan tinggi ke atas DNA dan RNA bagi pembahagian sel. Kajian ini bertujuan untuk memekakan sel kanser terhadap BQR dengan menurunkan aktivitiUCK2. Dalam sel rebahUCK2 DLD-1, tiada sebarang perubahan kesensitifan sel terhadap BQR diperhatikan. Uridina telah diketahui dapat membalikkan kesan anti-pembahagian perencat DHODH melalui tapak jalan penyelamatan pirimida. Kami mendapati bahawa pembatalan kesan pembalikan uridina adalah lebih kurang 30% di dalam sel rebah UCK2 berbanding sel jenis liar. Penemuan kami menunjukkan bahawa kehilangan aktivitiUCK2 di dalam laluan penghematan tidak meningkatkan perencatan pembahagian sel berperantarakan BQR tetapi membatalkan pembalikan uridina di dalam sel.

Kata kunci: BQR; DHODH; pembatalan uridina; TAS-106; UCK2

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*Corresponding author; email: shatraho@um.edu.my