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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
Received: 2 April 2012/Accepted: 16
December 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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*Corresponding author; email: hafizrothi@salam.uitm.edu.my
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