Analisis Ekspresi Protein c-Fos pada Cell Line Kanker Payudara

Aisyah Elliyanti

Abstract

Tujuan: Untuk menganalisis ekspresi protein c-Fos pada cell line kanker payudara. Metode: Penelitian ini merupakan penelitian eksperimental murni menggunakan metode imunositofluoresens untuk melihat jumlah intensitas ekspresi protein c-Fos dengan sampel penelitian adalah cell line MCF-7 mewakili kanker payudara subtipe luminal A, cell line SKBR3 mewakili kanker payudara subtipe HER2+, dan cell line HaCat mewakili sel normal. Ekspresi protein c-Fos diinduksi dengan EGF 50 ng/mL, ATP 100µM dan gabungan ATP+EGF selama 45 menit. Perhitungan intensitas protein c-Fos menggunakan aplikasi imageJ. Hasil penelitian ekspresi c-Fos dianalisis menggunakan uji oneway anova  apabila p<0.05 dianggap berbeda secara signifikan. Hasil: Ekspresi protein c-Fos pada cell line yang diberi perlakuan ATP dan EGF meningkat dibandingkan kontrol. Peningkatan intensitas protein c-Fos terdapat pada ketiga jenis cell line. Tidak terdapat perbedaan bermakna pada ketiga jenis sel berdasarkan jenis induksi (p>0,05). Kesimpulan: Pemberian ATP, EGF dan kombinasi ATP+EGF meningkatkan intensitas protein c-Fos pada cell line kanker payudara tipe MCF-7 dan SKBR3.

Keywords

: ATP; EGF; Protein c-Fos; Intensitas

Full Text:

PDF

References

Warner E. Breast-Cancer Screening. N Eng J Med. 2011;365:1025–32.

Qodria L, Nurrachma MY. Pemilihan Sel yang Tepat Untuk Penelitian Kanker Payudara. BioTrends. 2020;11(2):17–28.

GLOBOCAN. The Global Cancer Observatory - All cancers. Int Agency Res Cancer - WHO. 2020;419:199–200.

The Global Cancer Observatory. Cancer Incident in Indonesia. Int Agency Res Cancer. 2020;858:1–2.

Warjianto W, Soewoto W, Alifianto U, Wujoso H. Hubungan Reseptor Estrogen, Reseptor Progesteron dan Ekspresi Her-2/Neu Dengan Grading Histopatologi pada Pasien Kanker Payudara di RSUD dr. Moewardi Surakarta. Smart Med J. 2020;3(2):96.

Arnetha TS, Hernowo BS, Adha MJ, Rezano A. Relationship between Molecular Subtypes and Overall Survival of Breast Cancer in Bandung. Biomed Pharmacol J. 2020;13(3):1543–8.

Ramaswamy MZ and B. Mechanisms and therapeutic advances in the management of endocrine-resistant breast cancer. World J Clin Oncol. 2014;5(3):248–62.

Elliyanti A. Peran C-Fos Sebagai Agen Proliferasi Dan Pro-Apoptosis Sebagai Strategi Pengembangan Pengobatan Kanker. Maj Kedokt Andalas. 2016;39(2):73.

Mahner S, Baasch C, Schwarz J, Hein S, Wölber L, Jänicke F, et al. C-Fos expression is a molecular predictor of progression and survival in epithelial ovarian carcinoma. Br J Cancer. 2008;99(8):1269–75.

Nakakuki T, Birtwistle MR, Saeki Y, Yumoto N, Ide K, Nagashima T, et al. Ligand-specific c-fos expression emerges from the spatiotemporal control of ErbB network dynamics. Cell. 2010;141(5):884–96.

Mikula M, Gotzmann J, Fischer ANM, Wolschek MF, Thallinger C, Schulte-Hermann R, et al. The proto-oncoprotein c-Fos negatively regulates hepatocellular tumorigenesis. Oncogene. 2003;22(42 REV. ISS. 4):6725–38.

Seon Pil Jin, Ji Hun Kim, Min A Kim, Han-Kwang Yang, Hee Eun Lee HSL& WHK. Prognostic significance of loss of c-fos protein in gastric carcinoma. Pathol Oncol Res. 2007;13:284–89.

Elliyanti A et al. Correlation Between Natrium Iodide Symporter and c-Fos Expression in Breast Cancer Cell Line Advances in Biomolecular Medicine. CRC Press. 2017;19–22.

Breast cancer [Internet]. Cancer Research UK. [cited 2022 Apr 23]. Available from: https://www.cancerresearchuk.org/about-cancer/breast-cancer

Ferguson FMAT. Breast Cancer. ncbi. 2021;70(8):515–7.

Watkins E. Overview of breast cancer. J Am Acad Physician Assist. 2019;32(10):13–7.

Indonesia Departemen Kesehatan Republik. Riset Kesehatan Dasar 2018. 2019.

Hasnita Y, Harahap WA, Defrin. Penelitian Pengaruh Faktor Risiko Hormonal pada Pasien Kanker Payudara di RSUP. Dr. M. Djamil Padang. J Kesehat Andalas. 2019;8(3):522–8.

Putu Nita Cahyawati. Imunoterapi pada Kanker Payudara. WICAKSANA. 2018;2(1).

Breast cancer symptoms [Internet]. Cancer Research UK. [cited 2022 Apr 24]. Available from: https://www.cancerresearchuk.org/about-cancer/breast-cancer/symptoms

Indonesia KMKR. Pedoman Nasional Pelayanan Kedokteran Tata Laksana Kanker Payudara. 2018;2(2):2016.

Tests to stage breast cancer [Internet]. Cancer Research UK. [cited 2022 Apr 24]. Available from: https://www.cancerresearchuk.org/about-cancer/breast-cancer/getting-diagnosed/tests-stage

Sari SE, Harahap WA, Saputra D. Pengaruh Faktor Risiko Terhadap Ekspresi Reseptor Estrogen Pada Penderita Kanker Payudara Di Kota Padang. J Kesehat Andalas. 2018;7(4):461.

Tsang JYS, Tse GM. Molecular Classification of Breast Cancer. Adv Anat Pathol. 2020;27(1):27–35.

Torsten O Nielsen, Samuel C. Y Leung, David L Rimm AD et al. Assessment of Ki67 in Breast Cancer: Updated Recommendations From the International Ki67 in Breast Cancer Working Group. J Natl Cancer Inst. 2021;113(7):808–19.

Feng Y, Spezia M, Huang S, Yuan C, Zeng Z, Zhang L, et al. Breast cancer development and progression: Risk factors, cancer stem cells, signaling pathways, genomics, and molecular pathogenesis. Genes Dis. 2018;5(2):77–106.

Treatment for Breast Cancer [Internet]. Cancer Research UK. [cited 2022 Apr 24]. Available from: https://www.cancerresearchuk.org/about-cancer/breast-cancer/treatment/treatment-decisions

Collins Dictionary of Medicine [Internet]. HarperCollins Publishers. 2014 [cited 2022 Apr 12]. Available from: https://www.thefreedictionary.com/cell line

Smith1† SE, Mellor1† P, Ward1† AK, Kendall1† S, McDonald2 M, Vizeacoumar1 FS, et al. Molecular characterization of breast cancer cell lines through multiple omic approaches. Breast Cancer Res. 2017;19(65).

Dai X, Cheng H, Bai Z, Li J. Breast cancer cell line classification and Its relevance with breast tumor subtyping. J Cancer. 2017;8(16):3131–41.

Ayu MS. Uji Sitotoksitas dan Proliferasi Senyawa 1-(2-Klorobenzoiloksimetil)-5-Fluorourasil terhadap Sel kanker Payudara (sel MCF-7). 2015.

Comsa S, Maria A MR. The Story of MCF-7 Breast Cancer Cell Line: 40 years of Experience in Research. Anticancer Res. 2015;35(6):3147–54.

MCF-7 [Internet]. ATCC (American Tissue Culture Collection). [cited 2022 Apr 23]. Available from: https://www.atcc.org/products/htb-22

SKBR3 [Internet]. ATCC (American Tissue Culture Collection). [cited 2022 Apr 23]. Available from: https://www.atcc.org/products/htb-30

SKBR-3: Human Breast Cancer Cell Line (ATCC HTB-30) [Internet]. Memorial Sloan Kettering Cancer Center. 2022 [cited 2022 Apr 22]. Available from: https://www.mskcc.org/research-advantage/support/technology/tangible-material/human-breast-cell-line-sk-br-3

MDA-MB-231 Cell line profile. ECACC, Eur Collect Authenticated Cell Cult. 2022;4(92020424):1–2.

MDAMB231 [Internet]. ATCC (American Tissue Culture Collection). [cited 2022 Apr 23]. Available from: https://www.atcc.org/products/htb-26

Wilson AFD& VG. In vitro culture conditions to study keratinocyte differentiation using the HaCaT cell line. Cytotechnology. 2007;54(1):77–83.

Chia-Lin Ho, Chih-Yung Yang, Wen-Jie Lin C-HL. Ecto-Nucleoside Triphosphate Diphosphohydrolase 2 Modulates Local ATP-Induced Calcium Signaling in Human HaCaT Keratinocytes. Plosone. 2013;1371.

Accepted MCB, Posted M, Society A, Reserved AR. Evidence for Homodimerization of the c-Fos Transcription Factor in Live Cells Revealed by Fluorescence Microscopy and Computer Modeling. Mol Cell Biol. 2015;35(21):3785–98.

Mahner S, Baasch C, Schwarz J, Hein S, Wölber L, Jänicke F, et al. C-fos Expression is Molecular Predictor Progression. BJC. 2008. p. 1269–75.

UniProtKB - P01100 (FOS_HUMAN) [Internet]. UniProt. [cited 2022 Apr 24]. Available from: https://www.uniprot.org/uniprot/P01100

Bao-Sheng Chen, Ming-Rong Wang, Yan Cai, Xin Xu, Zhi-Xiong Xu, Ya-Ling Han MW. Decreased expression of SPRR3 in Chinese human oesophageal cancer. Carcinogenesis. 2000;21(12):2147–50.

Wagstaff SC, Bowler WB, Gallagher JA, Hipskind RA. Extracellular ATP activates multiple signalling pathways and potentiates growth factor-induced c-fos gene expression in MCF-7 breast cancer cells. Carcinogenesis. 2000;21(12):2175–81.

Pacheco-pantoja EL, Dillon JP, Wilson PJM, Fraser WD, Gallagher JA. c-Fos induction by gut hormones and extracellular ATP in osteoblastic-like cell lines. Purinergic Signal. 2016;7–11.

Elliyanti A, Putra AE, Sribudiani Y, Noormartany N, Masjhur JS, Achmad TH, et al. Epidermal growth factor and adenosine triphosphate induce natrium iodide symporter expression in breast cancer cell lines. Open Access Maced J Med Sci. 2019;7(13):2088–92.

Publications S. Immunofluorescence Assays. ibidi. 2019;10(1038):419–67.

Odell ID, Cook D. Immunofluorescence Techniques. J Invest Dermatol. 2013;133(1):1–4.

Farhana. MAA. Enzyme Linked Immunosorbent Assay. ncbi. 2022;657–82.

Santosa B. Teknik Elisa. Semarang: Unimus Press; 2020. 36 p.

Syennie Sari Agung, Iman Permana Maksum TS. Serum Otologus dan human Epidermal Growth Factor (hEGF) Mempercepat Proliferasi dan Migrasi Keratinosit pada Proses Re-Epitelisasi. MKB. 2016;48(4).

Abcam. ab264626 Human c-Fos SimpleStep ELISA kit. In: Abcam. 2021. p. 24.

Elliyanti A, Wikayani TP, Masjhur JS, Achmad TH. Deteksi Natrium / Iodide Symporter ( NIS ) pada Galur Sel Kanker Payudara SKBR3 dengan Imunositofluoresens Detection of Natrium / Iodide Symporter ( NIS ) in SKBR-3 Breast Cancer Cell Line Using Immunocytofluoresence. 2014;48(1):15–8.

Jimeno A, Kulesza P, Kincaid E, Bouaroud N, Chan A, Forastiere A et al. Cfos Assesment as A Marker of Anti-epidermal Growth Factor Receptor Effect. Cancer Res. 2006;(66):2385–90.

E Garcia, D Lacasa YG. Estradiol stimulation of c-fos and c-jun expressions and activator protein-1 deoxyribonucleic acid binding activity in rat white adipocyte. Endocrinology. 2000;141(8):2837–46.

C.Partridge AT-YDBRP. Parathyroid Hormone Induces c-fos Promoter Activity in Osteoblastic Cells through Phosphorylated cAMP Response Element (CRE)-binding protein Binding to the Major CRE. J Biol Chem. 1996;271(41):25715–21.

Budiarto BR. Polymerase Chain Reaction (PCR) : Perkembangan Dan Perannya Dalam Diagnostik Kesehatan. BioTrends. 2015;6(2):29–38.

Rezgar Rahbari, Nariman Moradi and MA. rRT-PCR for SARS-CoV-2: Analytical considerations. Clin Chim Acta. 2021;1(516):1–7.

Mils Â, Piette J, Barette C, Veyrune J, Tesnie A, Escot C, et al. The proto-oncogene c-fos increases the sensitivity of keratinocytes to apoptosis The proto-oncogene c- fos increases the sensitivity of keratinocytes to apoptosis. 1997;(May).

Xu Y, Voorhees JJ, Fisher GJ. Epidermal Growth Factor Receptor Is a Critical Mediator of Ultraviolet B Irradiation-Induced Signal Transduction in Immortalized Human Keratinocyte HaCaT Cells. Am J Pathol. 2006;169(3):823–30.

Jennifer L. Hsu, Mien-Chie Hung. The role of HER2, EGFR, and other receptor tyrosine kinases in breast cancer. Cancer Metastasis Rev. 2016;35(4)575-88.



-->