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Sains
Malaysiana 46(4)(2017): 575–581 http://dx.doi.org/10.17576/jsm-2017-4604-09 Low Intensity Ultrasound Induced
Apoptosis in MCF-7 Breast Cancer Cell Lines (Ultrabunyi
Keamatan Rendah Aruhan Apoptosis dalam Jujukan Sel Kanser Payudara
MCF-7) SITI
P.M.
BOHARI*,
HAMIDREZA
ABOULKHEYR,
E.S.,
NUR
S.
JOHAN
& NURSYUHADA
F.
ZAINUDIN Faculty of Biosciences
and Biomedical Engineering, Universiti Teknologi Malaysia 81310 UTM Skudai, Johor
Darul Takzim, Malaysia Diserahkan: 26 November
2015/Diterima: 29 September 2016 ABSTRACT According
to the World Cancer Research Fund International (WCRFI),
breast cancer is the most common type of cancer in women worldwide
with recorded 1.7 million new cases in 2012. The main line of treatments
is still limited to chemotherapy, surgery and radiotherapy which
could lead to a wide range of dangerous side effects. This study
was conducted to evaluate the effect of low intensity ultrasound
(LIUS) on cell proliferation, percentage
of living and dead cells and the induction of apoptosis on the MCF-7
cell line with CHO cells as the control for non-cancerous
group. In order to achieve the objective of this study, several
methods of cell-bioguided assays were used including the MTT assay
for cell proliferation, Live/Dead assay for the determination of
both live and dead cells and gene expression study for the detection
of apoptosis in the cells. The cytotoxicity and Live/Dead assays
data provided preliminary data that the LIUS has potential to induce apoptosis in a wide population
of breast cancer cells. Furthermore, the LIUS treatment
induced the expression of p53-mRNA at a detectable level via
qPCR
analysis, indicating the activation of apoptosis.
In short, our study suggested LIUS
dosage used in this study could potentially show positive
effects in the induction of apoptosis selectively on the MCF-7
with less harm to the control CHO cells. Keywords:
Apoptosis; CHO cells; LIUS; MCF-7
cells; qPCR ABSTRAK Mengikut ‘World
Cancer Research Fund International’ (WCRFI),
kanser payudara adalah jenis kanser yang paling umum pada wanita
di seluruh dunia dengan catatan rekod 1.7 juta kes baru pada 2012.
Rawatan utama masih lagi terhad kepada kimoterapi, pembedahan dan
radioterapi yang boleh membawa kepada pelbagai kesan sampingan berbahaya.
Penyelidikan ini dijalankan untuk menilai kesan ultrabunyi berkeamatan
rendah (LIUS)
pada pertumbuhan sel, peratusan hidup dan mati sel dan juga pengaruhan
‘apoptosis’ pada jujukan sel MCF-7 dan CHO sebagai
kumpulan kawalan sel bukan kanser. Untuk mencapai objektif di dalam
kajian ini, beberapa kaedah bioasai berdasarkan sel telah digunakan
termasuklah asai MTT
untuk pertumbuhan sel, asai ‘Live/Dead’ untuk pengenalpastian
hidup dan mati sel dan juga kajian ekspresi gen untuk pengesanan
‘apoptosis’ pada sel. Asai sitotoksiti dan ‘Live/Dead’ menggambarkan
data awal bahawa LIUS berpotensi
untuk mengaruh ‘apoptosis’ di dalam populasi yang besar bagi kanser
payudara. Tambahan lagi, rawatan LIUS juga berpotensi untuk mengaruh ekspresi
p53-mRNA pada aras yang dapat dikesan melalui analisis qPCR
yang menunjukkan pengaktifan ‘apoptosis. Kesimpulannya,
kajian kami mencadangkan dos LIUS
yang digunakan dalam kajian ini berpotensi dalam menunjukkan
kesan positif dalam pengaruhan ‘apoptosis’ pilihan pada sel MCF-7
dengan sedikit kemudaratan kepada sel CHO. Kata kunci: Apoptosis; LIUS; qPCR;
sel CHO; sel MCF-7 RUJUKAN Bohari, S.P.M., Hukins, D.W.L. & Grover, L.M. 2011. Effect of
calcium alginate concentration on viability and proliferation of
encapsulated fibroblasts. Bio-Medical Materials and Engineering
21(3): 159-170. Cambier, D., D’Herde, K., Witvrouw, E., Beck, M., Soenens, S. &
Vanderstraeten, G. 2001. Therapeutic ultrasound: Temperature increase
at different depths by different modes in a human cadaver. Journal
of Rehabilitation Medicine 33(5): 212-215. Chumakova, O.V., Liopo, A.V., Evers, B.M. & Esenaliev, R.O. 2006.
Effect of 5-fluorouracil, optison and ultrasound on MCF-7 cell viability.
Ultrasound in Medicine & Biology 32(5): 751-758. Dubinsky, T.J., Cuevas,
C., Dighe, M.K., Kolokythas, O. & Hwang, J.H. 2008. High-intensity
focused ultrasound: Current potential and oncologic applications.
American Journal of Roentgenology 190(1): 191-199. Feng, Y., Tian, Z. &
Wan, M. 2010. Bioeffects of low-intensity ultrasound in vitro
apoptosis, protein profile alteration, and potential molecular
mechanism. Journal of Ultrasound in Medicine 29(6): 963-974.
Garg, A.K. & Buchholz,
T.A. 2015. Influence of neoadjuvant chemotherapy on radiotherapy
for breast cancer. Annals of Surgical Oncology 22(5): 1434-1440.
Guo, W. 2014. Concise
review: Breast cancer stem cells: Regulatory networks, stem cell
niches, and disease relevance. Stem Cells Translational Medicine
3(8): 942-948. Hortobagyi, G.N., de
la Garza Salazar, J., Pritchard, K., Amadori, D., Haidinger, R.,
Hudis, C.A., Hussein Khaled; Liu, M-C., Martin, M. & Namer,
M. 2005. The global breast cancer burden: Variations in epidemiology
and survival. Clinical Breast Cancer 6(5): 391-401. Hui, L., Zheng, Y., Yan,
Y., Bargonetti, J. & Foster, D.A. 2006. Mutant p53 in MDA-MB-231
breast cancer cells is stabilized by elevated phospholipase D activity
and contributes to survival signals generated by phospholipase D.
Oncogene 25(55): 7305-7310. Hupp, T., Lane, D. &
Ball, K. 2000. Strategies for manipulating the p53 pathway in the
treatment of human cancer. Biochem. J. 352: 1-17. Johns, L.D. 2002. Nonthermal
effects of therapeutic ultrasound: The frequency resonance hypothesis.
Journal of Athletic Training 37(3): 293-299. Komen, J., Wolbers, F.,
Franke, H.R., Andersson, H., Vermes, I. & van den Berg, A. 2008.
Viability analysis and apoptosis induction of breast cancer cells
in a microfluidic device: Effect of cytostatic drugs. Biomedical
Microdevices 10(5): 727-737. Larina, I.V., Evers,
B.M. & Esenaliev, R.O. 2005. Optimal drug and gene delivery
in cancer cells by ultrasound-induced cavitation. Anticancer
Research 25(1A): 149-156. Li, Y., Wang, P., Zhao,
P., Zhu, S., Wang, X. & Liu, Q. 2012. Apoptosis induced by sonodynamic
treatment by protoporphyrin IX on MDA-MB-231 cells. Ultrasonics
52(4): 490-496. Luo, L., Molnar, J.,
Ding, H., Lv, X. & Spengler, G. 2006. Ultrasound absorption
and entropy production in biological tissue: A novel approach to
anticancer therapy. Diagnostic Pathology 1(1): 35. Ninomiya Kazuaki, Takahiro
Yamashita, Shinya Kawabata & Nobuaki Shimizu. 2014. Targeted
and ultrasound-triggered drug delivery using liposomes co-modified
with cancer cell-targeting aptamers and a thermosensitive polymer.
Ultrasonics Sonochemistry 21(4): 1482-1488. Paliwal, S. & Mitragotri,
S. 2008. Therapeutic opportunities in biological responses of ultrasound.
Ultrasonics 48(4): 271-278. Pallant, J. 2007. SPSS
Survival Manual. 3rd ed. Berkshire: McGraw-Hill. Park, S.M., Kim, M.S.,
Park, S-J., Park, E.S., Choi, K-S., Kim, Y-S. & Kim, H.R. 2013.
Novel temperature-triggered liposome with high stability: Formulation,
in vitro evaluation, and in vivo study combined with
high-intensity focused ultrasound (HIFU). Journal of Controlled
Release 170(3): 373-379. Shibaguchi Hirotomo,
Hirofumi Tsuru, Motomu Kuroki & Masahide Kuroki. 2011. Sonodynamic
cancer therapy: A non-invasive and repeatable approach using low-intensity
ultrasound with a sonosensitizer. Anticancer Research 31(7):
2425-2429. Tsao, H., Zhang, X.,
Majewski, P. & Haluska, F.G. 1999. Mutational and expression
analysis of the p73 gene in melanoma cell lines. Cancer Research
59(1): 172-174. Uludag, H. & Sefton,
M.V. 1990. Colorimetric assay for cellular-activity in microcapsules.
Biomaterials 11(9): 708-712. Wang, H., Wang, X., Wang,
P., Zhang, K., Yang, S. & Liu, Q. 2013. Ultrasound enhances
the efficacy of chlorin E6- mediated photodynamic therapy in MDA-MB-231
cells. Ultrasound in Medicine & Biology 39(9): 1713-1724.
Watanabe, Akihiro, Takeshi
Takatera, Toshio Sato, S Takeuchim, Hiroyuki Nishimura & Norimichi
Kawashima. 2002. Study on suppression mechanism of cancer cells
proliferation by ultrasound exposure for minimally invasive cancer
treatment. Paper read at Ultrasonics Symposium, 2002. Proceedings.
IEEE. Wood, A.K.W. & Sehgal,
C.M. 2015. A review of low-intensity ultrasound for cancer therapy.
Ultrasound in Medicine & Biology 41(4): 905-928. Wu, J. & Nyborg,
W. 2006. Emerging Therapeutic Ultrasound. Singapore: World
Scientific Publishing Co. Pte. Ltd. *Pengarang untuk surat-menyurat; email: pauliena@utm.my
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