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Sains Malaysiana 42(8)(2013):
1131–1137
Targeted RNAi of the Mitogen-activated Protein Kinase Pathway Genes in Acute Myeloid Leukemia Cells
(RNAi Sasar Gen Tapak Jalan Protein Kinase Diaktifkan-Mitogen
dalam Mieloid Leukemia Akut)
M.R.
Mohd Hafiz1
1 Malaysia
2
Diserahkan:
2 April 2012/Diterima: 16 Disember 2012
ABSTRACT
In this study, RNA interference (RNAi) was carried out as an
experimental technique to knockdown three mitogen-activated protein kinase (MAPK)
pathway genes, raf-1, mekk1 and mlk3 in acute myeloid leukemia (AML) cells.
Conventionally, RNAi knockdown experiments target a single gene for functional
studies or therapeutic purposes. We wanted to explore the potential differences
or similarities between targeting single targets or multiple target genes in a
single application. We achieved knockdown of gene expression levels of between
40 and 60% for the RNAi experiments, with better knockdown observed in single
target gene experiments in comparison with the multiple target gene experiment.
Microarray analysis indicated that the transfection process had most likely
induced the immune response from the cells in every RNAi treatment. This might
indicate that when the MAPK signaling pathway is partially blocked,
in tandem with the immune response, the cells will begin signaling for
apoptosis leading to cellular death of the leukemic cells.
Keywords: Acute myeloid leukemia; immune response; MAPK pathway; RNA interference
ABSTRAK
Dalam kajian ini, penggangguan RNA (RNAi) digunakan
sebagai teknik uji kaji untuk menurunkan tiga gen protein kinase
diaktifkan-mitogen (MAPK) iaitu gen raf1, mekk1 dan mlk3 di dalam
sel mieloid leukemia akut (AML). Kebiasaannya,
eksperimen RNAi dijalankan untuk menyasar satu gen sahaja demi mengkaji fungsi
atau peranan terapi. Kami telah mengkaji potensi perbezaan atau
persamaan antara menyasar satu atau lebih gen dalam satu aplikasi. Kami berjaya
mencapai penurunan pengekspresan gen daripada 40% hingga 60% dan RNAi kelihatan
lebih berkesan melalui penyasaran satu gen. Analisis mikroatur menunjukkan
bahawa proses transfeksi kemungkinan tinggi telah mengaruh tindak balas imun
dalam setiap perlakuan RNAi yang telah dilakukan. Ini mungkin memberi petunjuk
bahawa apabila pengisyaratan tapak jalan MAPK dihalang separa, disertakan pengaruhan tindak
balas imun, tapak laluan apoptosis akan dimulakan dan
mengakibatkan kematian sel kepada sel-sel leukemia.
Kata kunci: Mieloid leukemia akut; penggangguan RNA;
tapak jalan MAPK; tindak balas imun
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*Pengarang untuk surat-menyurat; email: hafiz.rothi@gmail.com
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