Sains Malaysiana 51(1)(2022): 175-186

http://doi.org/10.17576/jsm-2022-5101-14

P2Y Purinergic Receptor Signaling in Oral Squamous Cell Carcinoma Cell Lines and Its Role in Proliferation and Cisplatin-Mediated Apoptosis

(Isyarat Reseptor Purinergik P2Y dalam Titisan Sel Oral Skuamus Sel Karsinoma dan Peranannya dalam Proliferasi dan Apoptosis Didorong Cisplatin)

LOK MUN LAW¹, NORAZRINA AZMI¹, IAN CHARLES PATERSON^2,3 & PEI YUEN NG¹*

¹Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

²Department of Oral & Craniofacial Science, Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

³Oral Cancer Research and Coordinating Centre, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

Received: 29 August 2020/Accepted: 23 April 2021

ABSTRACT

Treatment of advanced stage oral squamous cell carcinoma (OSCC) often involves the use of chemotherapeutic agents, such as cisplatin. However, its use often results in therapeutic failure due to chemoresistance. This study focused on a class of purinergic receptors, namely P2Y, which are activated via interaction with extracellular nucleotides. The functional effects of P2Y receptor activation in OSCC cell lines as well as the signaling pathways involved were investigated. The expression of P2Y₂ receptors in histological sections of OSCC was studied due to its association with cancer. Activation of MAPK pathways via extracellular nucleotides were studied in OSCC cell lines, along with downstream effects such as proliferation and cisplatin-mediated apoptosis. Immunohistochemical staining of OSCC tissue samples showed loss of P2Y₂ expression as the disease progressed. Western blotting identified different MAPK signaling pathways were activated by extracellular nucleotides. Bromodeoxyuridine proliferation assays showed increased cellular proliferation in the OSCC cell lines H400 (p < 0.001) and SAS (p < 0.001) after 24 h treatment with ATP. However, the ability of extracellular nucleotides to activate multiple P2Y receptor subtypes may indicate the involvement of other subtypes aside from P2Y₂. Cisplatin-mediated apoptosis was enhanced in SAS cells co-treated with ATP (p < 0.001), while H376 (p < 0.001) showed reduction in the number of apoptotic cells and no significant changes were observed in H103. This study concluded that extracellular nucleotide on OSCC cell lines with different characterizations had varied downstream effects, which suggests the use of targeted therapy to specific individuals.

Keywords: Adenosine triphosphate; extracellular nucleotide; oral squamous cell carcinoma; P2Y; purinergic receptor

ABSTRAK

Rawatan karsinoma sel skuamus mulut (OSCC) peringkat lanjut selalunya melibatkan penggunaan agen kemoterapi seperti cisplatin. Akan tetapi, rawatan sering kali gagal disebabkan oleh kerintangan terhadap agen kemoterapi tersebut. Kajian ini menumpukan perhatian pada satu kelas reseptor purinergik iaitu P2Y kerana ia diaktifkan oleh interaksi dengan nukleotida ekstrasel. Pengekspresan reseptor P2Y₂ pada OSCC ditentukan dalam kajian histologi kerana ia sering dikaitkan dengan kanser. Pengaktifan tapak jalan MAPK oleh nukleotida ekstrasel juga dikaji dalam titisan sel OSCC, bersama dengan kesan hiliran seperti proliferasi dan apoptosis oleh cisplatin. Pewarnaan imunohistokimia menunjukkan penurunan pengekspresan P2Y₂ dengan perkembangan penyakit. Pemblotan Western juga menunjukkan pengaktifan tapak jalan MAPK yang berlainan dengan nukleotida yang digunakan. Ujian proliferasi juga menunjukkan peningkatan kadar proliferasi pada titisan sel OSCC H400 dan SAS selepas 24 jam rangsangan oleh nukleotida ekstrasel. Akan tetapi, kebolehan nukleotida ekstrasel untuk berinteraksi dengan golongan subjenis reseptor P2Y berkemungkinan menunjukkan kesan yang dikaji melibatkan reseptor selain daripada P2Y₂. Apoptosis oleh cisplatin ditingkatkan dalam titisan sel SAS selepas rangsangan nukleotida ekstrasel, manakala titisan sel H376 pula menunjukkan penurunan bilangan sel apoptosis. Hasil kajian ini mendapati bahawa titisan sel OSCC yang berlainan dari segi penciriannya mempunyai kesan hiliran yang berbeza dan ini mencadangkan rawatan khusus yang disasarkan terhadap individu.

Kata kunci: Adenosina trifosfat; karsinoma sel skuamus mulut; nukleotida ekstrasel; P2Y; reseptor purinergik

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*Corresponding author; email: pyng@ukm.edu.my