Sains Malaysiana 50(9)(2021): 2701-2711

http://doi.org/10.17576/jsm-2021-5009-17

 

Ranibizumab Inhibits Human Tenon’s Fibroblast Proliferation via p21 Dependent p53 Mechanisms

(Ranibizumab Merencat Percambahan Fibroblas Tenon Manusia melalui Mekanisme p21 Bersandar p53)

 

 SITI MUNIRAH MD NOH1*, SITI HAMIMAH SHEIKH ABDUL KADIR2,3 & SUSHIL VASUDEVAN2

 

1University of Malaya Centre of Innovation and Commercialization (UMCIC), University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

 

2Faculty of Medicine, Universiti Teknologi MARA, Cawangan Sungai Buloh, 47000 Sungai Buloh, Selangor Darul Ehsan, Malaysia

 

3Institute for Pathology, Laboratory and Forensic Medicine (I-PPerForM), Universiti Teknologi MARA, Cawangan Sungai Buloh, 47000 Sungai Buloh, Selangor Darul Ehsan, Malaysia

 

Diserahkan: 11 Ogos 2020/Diterima: 19 Januari 2021

 

ABSTRACT

Trabeculectomy is the gold standard procedure performed in glaucoma when topical medication and laser intervention failed. In a trabeculectomy, number of clinical trials have shown the efficacy of ranibizumab in minimizing extracellular matrix accumulation at the filtering site. Ranibizumab (LucentisTM) is a drug that targets vascular endothelial growth factor (VEGF). However, to date the actual mechanisms of this anti-VEGF in trabeculectomy is still not well understood. Hence, in here we aimed to elucidate the effects of ranibizumab on human Tenon’s fibroblast (HTF) isolated from patients undergoing trabeculectomy. In our previous study, we had reported that ranibizumab reduces the level of spermidine metabolite whereby spermidine is an important polyamine for cell proliferation. For this current study, cultured HTFs were divided into untreated, control IgG, ranibizumab only, difluoromethylornithine (DFMO; inhibitor of spermidine) only and ranibizumab with DFMO. All cells were extracted for PCR array (expression of CDKN1A, CDK2, and CDK4) and protein expression of p53, p21, CDK2, and CDK4 by Western Blot. In here, our result demonstrated that cells treated with ranibizumab or DFMO and cells treated with ranibizumab-DFMO have similar effects as both show increased in p53 and p21. Meanwhile, no significant differences in expression of CDKN1A, CDK2 and CDK4 were observed in all groups. In essence, our findings suggest that ranibizumab action is mediated by p21 and p53.

 

Keywords: Human Tenon’s fibroblast; ranibizumab; vascular endothelial growth factor

 

ABSTRAK

Trabekulektomi adalah prosedur utama yang dijalankan bagi penyakit glaukoma apabila ubatan topikal dan intervensi laser telah gagal. Dalam trabekulektomi, beberapa ujian klinikal telah menunjukkan keberkesanan ranibizumab dalam menurunkan pengumpulan metriks ekstrasel pada tapak penurasan. Ranibizumab (LucentisTM) adalah sejenis dadah yang mensasarkan faktor pertumbuhan endotelium vaskular (VEGF). Namun, sehingga kini masih belum difahami sepenuhnya bagaimana anti-VEGF bekerja dalam mengurangkan kesan sampingan bagi trabekulekomi. Oleh kerana itu, kami mensasarkan untuk mengkaji kesan ranibizumab ke atas fibroblas Tenon manusia (HTF) yang telah diambil daripada pesakit yang melalui prosedur trabekulektomi. Dalam kajian lepas, kami telah melaporkan bahawa ranibizumab menurunkan kadar metabolit spermidin, iaitu sejenis poliamina penting untuk pertumbuhan sel. Dalam kajian semasa ini, kultur HTFs dibahagikan kepada tanpa rawatan, IgG kawalan, ranibizumab, difluorometilornitin (DFMO; perencat spermidin) dan ranibizumab bersama DFMO. Semua sel diekstrak untuk tatasusunan PCR (ekspresi CDKN1A, CDK2 dan CDK4) dan ekspresi protein p53, p21, CDK2 dan CDK4 secara pemblotan western. Keputusan menunjukkan sel yang dirawat ranibizumab atau DFMO serta ranibizumab dengan DFMO menyebabkan peningkatan ekspresi p53 dan p21. Sementara, tiada perubahan signifikan dilihat bagiCDKN1A, CDK2 dan CDK4 dalam semua sel. Secara ringkasnya, keputusan kami mencadangkan bahawa ranibizumab mungkin bertindak melalui protein p21 dan p53.

 

Kata kunci: Faktor pertumbuhan endotelium vaskular; fibroblas Tenon manusia; ranibizumab

 

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*Pengarang untuk surat-menyurat; email: sitimunirah.umcic@um.edu.my  

 

 

   

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