Sains Malaysiana 46(8)(2017): 1289–1297
http://dx.doi.org/10.17576/jsm-2017-4608-15
A Gain of
Function p53 Gene Mutant Promotes Growth Suppression in Human Liver
Cancer Cells
(Pemulihan Fungsi Mutan Gen p53 Menggalakkan Penindasan Pertumbuhan Sel Kanser Hati Manusia)
NOR ADZIMAH JOHDI*, SITI NURMI NASIR
& RAHMAN JAMAL
UKM Medical Molecular
Biology Institute, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia
Diserahkan: 18 Ogos 2016/Diterima: 11 Januari 2017
ABSTRACT
Primary liver cancer
is one of the most common cancer in the world with
highest cancer mortality rate. The most common type of primary liver cancer is
hepatocellular carcinoma (HCC). There are many risk factors for
liver cancer and currently available treatments for HCC are
largely inadequate. Gene mutation and dysfunction of p53 are common and is recognized as an important molecular
event in hepatocarcinogenesis. Therefore, replacement
of the aberrant p53 gene is an attractive approach in the treatment of HCC providing
an alternative treatment for primary HCC. In this study, we assessed
whether the transfection with wild-type p53 gene is able to restore the
pro-apoptotic effects and evaluate the feasibility of gene therapy in fixing a
faulty p53 molecule. We established a non-viral cationic lipid-based p53 gene delivery into two human HCC cell lines namely HLF and PLC/PRF/5
cells. Both cell lines have mutations in the p53 gene. We compared the
results with the normal liver cell line, WRL68,
that constitutively expresses the wild-type p53 gene. In this
study, the introduction of wild-type p53 gene into HLF and PLC/PRF/5
cells resulted in an increased of p53 gene expression, protein expression
and cells growth inhibition shown in MTS reduction cell viability
assay, FITC-Annexin V and PI apoptosis
assay, western blot and caspase activity assay. In summary, the study provides
a promising therapeutic approach for p53 gene delivery into HCC patients.
The p53 gene delivery can be instituted together with chemotherapy as a
combination treatment to induce apoptosis.
Keywords: Apoptosis;
cell lines; hepatocellular carcinoma; p53 gene;
transfection
ABSTRAK
Kanser hati utama adalah salah satu kanser umum di dunia dengan kadar
kematian kanser tertinggi. Jenis yang paling biasa untuk kanser
hati utama adalah karsinoma hepatosel (HCC). Terdapat banyak faktor
risiko untuk kanser hati utama dan rawatan untuk HCC sebahagian
besarnya tidak mencukupi. Mutasi gen dan kehilangan fungsi gen
p53 adalah faktor yang paling biasa dan gen p53 diiktiraf sebagai
antara rantaian molekul yang penting dalam proses pembentukan
kanser hati. Oleh itu, pemulihan mutan gen p53 yang defektif adalah
satu pendekatan yang menarik dalam rawatan HCC sebagai rawatan
alternatif untuk HCC utama. Dalam kajian ini, penilaian dilakukan
sama ada transfeksi gen p53 normal ke atas titisan sel HCC mampu
mengembalikan kesan pro-apoptotik dan potensi terapi gen dalam
pemulihan molekul p53 yang defektif. Ini melibatkan transfeksi
gen p53 normal menggunakan teknik lipid kationik kepada dua jenis
titisan sel HCC manusia iaitu HLF dan PLC/PRF/5. Kedua-dua jenis
titisan sel ini mempunyai mutasi dalam gen p53. Keputusan kajian
dibandingkan dengan titisan sel hati normal, WRL68, yang mengekspres
gen p53 normal. Transfeksi gen p53 normal ke dalam titisan sel
HLF dan PLC/PRF/5 menyebabkan peningkatan ekspresi gen p53, pertambahan
ekspresi protein p53 dan perencatan pertumbuhan titisan sel-sel
yang ditunjukkan dalam assai MTS sel pengurangan daya maju, assai
apoptosis FITC-Annexin V & PI, pemendapan barat dan aktiviti
caspase. Secara ringkasnya, kajian ini menyediakan pendekatan
terapeutik yang boleh digunakan untuk menyalurkan gen p53 ke dalam
sel pesakit HCC. Penyampaian gen p53 boleh dimulakan bersama dengan
kemoterapi sebagai rawatan kombinasi untuk mendorong aktiviti
apoptosis.
Kata kunci: Apoptosis; gen p53; hepatosel karsinoma; titisan sel; transfeksi
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*Pengarang untuk surat-menyurat; email: adzimah@ppukm.ukm.edu.my