Sains Malaysiana 40(11)(2011): 1247–1253

Differential Protein Expression in Senescent Human Skin Fibroblasts and Stress

Induced Premature Senescence (SIPS) Fibroblasts

(Perbezaan Pengekspresan Protein dalam Fibroblas Kulit Manusia Senesen dan Fibroblas

Senesen Pramatang Teraruh Tekanan)

Goon Jo Aan*, Haryati Ahmad Hairi, Suzana Makpol & Wan Zurinah Wan Ngah

Jabatan Biokimia, Fakulti Perubatan, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

Mariati Abdul Rahman

Jabatan Biologi Mulut Klinikal, Fakulti Pergigian, Universiti Kebangsaan Malaysia

50300 Kuala Lumpur, Malaysia

Saiful Anuar Karsani

Institut Sains Biologi, Fakulti Sains, Universiti Malaya, 50603 Kuala Lumpur, Malaysia

Received: 28 April 2010 / Accepted: 22 February 2011

ABSTRACT

Replicative senescence of human diploid fibroblasts (HDFs) occurs when cells lose their capacity to proliferate and enter a phase of irreversible growth arrest. Stress-induced premature senescence (SIPS) on the other hand is caused by subcytotoxic concentrations of various oxidants which trigger accelerated cellular senescence. In this study, a SIPS model was established by exposing human diploid fibroblasts (HDFs) to 20 μM H2O2 for two weeks. A proteomic comparison between young, senescent and SIPS cells was done using two dimensional gel electrophoresis (2DGE) to elucidate the changes in protein expression associated with cellular aging. Our analysis showed that 28 protein spots were differentially expressed in senescent cells whereas 10 protein spots were differentially expressed in SIPS as compared to young cells. Three similar protein spots were differentially expressed in both senescent and SIPS cells when compared to the young cells. These results indicate that a difference in protein expression exists between senescent cells and SIPS cells compared to young cells.

Keywords: Fibroblast; senescent cells; SIPS; two dimensional gel electrophoresis (2DGE)

ABSTRAK

Senesen replikatif fibroblas diploid manusia berlaku apabila sel hilang kapasiti untuk berproliferasi dan memasuki fasa pertumbuhan terencat secara tidak berbalik. Senesen pramatang teraruh tekanan (SIPS) pula berpunca daripada kepekatan subsitotoksik pelbagai oksidan yang boleh mengaruh senesen sel. Dalam kajian ini, model SIPS telah dihasilkan dengan mendedahkan fibroblas diploid manusia kepada 20 μM H2O2 selama dua minggu. Suatu perbandingan proteomik antara sel muda, sel tua dan model SIPS telah dilakukan dengan menggunakan elektroforesis gel dua dimensi (2DGE) untuk menunjukkan perubahan dalam pengekspresan protein berkaitan dengan penuaan selular. Keputusan menunjukkan 28 titik protein yang diekspreskan secara berbeza dalam sel senesen manakala SIPS menunjukkan 10 titik protein yang diekspreskan secara berbeza berbanding dengan sel muda. Tiga titik protein yang sama diekspreskan secara berbeza dalam kedua-dua sel senesen dan SIPS berbanding sel muda. Keputusan ini menunjukkan bahawa terdapat perbezaan dalam ekspresi protein antara sel senesen dan sel SIPS berbanding dengan sel muda.

Kata kunci: Elektroforesis gel dua dimensi; fibroblas; sel senesen; SIPS

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*Corresponding author; email: joaan@medic.ukm.my