PENGARUH VARIASI JUMLAH TEMBAKAN NANOSECOND PULSED ELECTRIC FIELDS (NsPEFs) TERHADAP EKSPRESI GEN SOCS3 PADA SEL KANKER SERVIKS HELA S3

Martina Kurnia Rohmah

Abstract


Abstrak

Nanosecond Pulsed Electric Fields (NsPEFs) merupakan teknologi bioelektrik yang berkembang dari teknologi elektroporasi. NsPEFs diberikan dengan intensitas tinggi namun dalam waktu yang sangat singkat yaitu 1 - 300 nanosekon. NsPEFs terbukti memiliki sejumlah efek biologis dan telah banyak dikembangkan dalam berbagai terapi salah satunya pada terapi kanker. Pada kanker serviks, protein HPV dapat menekan sejumlah ekspresi supresor tumor salah satunya yaitu gen Socs3. Penelitian ini bertujuan untuk mengetahui pengaruh perbedaan jumlah tembakan NsPEFs pada ekspresi gen Socs3. Sel HeLa S3 dikultur pada medium α-MEM dengan serum FBS 10%. Sebesar 20 kV/cm dalam durasi 80 ns NsPEFs dipapar pada suspensi sel dalam 4 mm cuvette. Gelombang NsPEFs dideteksi oleh probe bervoltase tinggi pada Oscilloscope. NsPEFs diberikan pada 0, 5, 10, 20, 30, 40, 0, dan 60 kali tembakan. Analisis ekspresi gen dilakukan dengan dua metode yaitu kuantitatif menggunakan Real time PCR dan kualitatif dengan RT-PCR. Data kuantitatif dianalisis secara statistik menggunakan Kolmogorf-Smirnov, Anova dan HSD Tukey (p<0.05). Hasil studi ini membuktikan bahwa paparan NsPEFs berpengaruh secara signifikan pada ekspresi gen Socs3 (p=0.000). Jumlah tembakan optimal 20 dan 30 kali dapat meningkatkan ekspresi gen Socs3 berturut-turut sebanyak = 2.779 dan = 3.105 kali. Ekspresi gen Socs3 akan menurun pada tembakan di atas 30 tembakan.

 

Kata Kunci: NsPEFs, tembakan, ekspresi, Socs3

 

Abstract

Nanosecond Pulsed Electric Fields (NsPEFs) is bioelectric that was developed by electroporation technology. NsPEFs use high intensity in short time exposure (1 – 300 nanosecond). NsPEFs have biological effect and was developed in cancer therapy. In cervical cancer, viral protein of HPV depresses some tumor suppressors like Socs3 gene. This research aims to investigate the effect of short variation in Socs3 gene expression. HeLa S3 cells were cultured in α-MEM with FBS 10%. NsPEFs as much as 20 kV/cm and 80 nano seconds was exposure over HeLa S3 cell in 4 mm cuvette. Wave of NsPEFs was detected by high voltage probe in oscilloscope. NsPEFs was exposure at 0 (control), 5, 10, 20, 30, 40, 50, and 60 shots. Socs3 gene expression was analyzed using real time PCR and RT-PCR. Quantitative data was analyzed by Kolmogorov-Smirnov, Anova, and HSD Tuker (p<0.05). This research show that NsPEFs is significantly increase Socs3 gene expression (p=0.000). The optimal shot 20 and 30 shots increase Socs3 gene expression subsequently = 2.779 and = 3.105 times. This expression decrease in higher than 30 shots of NsPEFs exposure.

 

Keywords: NsPEFs, shot, expression, Socs3


References


Beebe, S.J., Fox, P.M., Rec, L.J., Somers, K., Stark, R.H., Schoenbach, K.H. 2002. IEEE. Transact. Plasm. Sci. 30: 286-291.

Beebe, S.J., and Schoenbach, K.H. 2005. Nanosecond Pulsed Electric Fields: A New Stimulus to Activate Intracellular Signaling. J. Biomed. Biotech. 4: 297-300.

Beebe, S.J., Chen, Y.J., Sain, N.M., Schoenbach, K.H., Xiao, S. 2012. Transient Feature in Nanosecond Pulsed Electric Fields Differentially Modulate Mitochondria and Viability. PloS One. 7.

Beebe, S.J. 2013. Bioelectrics in Basic Science and Medicine: Impact of Electric Fields on Cellular Structures and Functions. Nanomed. and Nanotech. 4: 1-8.

Burd EM. Human Papillomavirus and Cervical Cancer. Clinical of Microbiology Review. 2003; 16(1): 1-17.

Chen C, Smye SW, Robinson MP, and Evan JA. 2006. Membrane Electroporation Theories: A Review. Medical & Biological Engineering & Computing. 44(1-2): 5-14.

Deng, J., Schoenbach, K.H., Buescher, E.S., Hair, P., Fox, P.M., Beebe, S.J. 2003. The Effect of Intense Submicrosecond Electrical Pulses on cells. Biophys. J. 84: 2709-2714.

Jimenez-Wences H, Peralta-Zaragoza O, and Fernández-Tilapa G. 2014. Human Papilloma Virus, DNA Methylation and micoRNA in Cervical Cancer. Oncology Report.. 31(6); 2467-2476.

Martin, A.G. 2007. Molecular Biology of Cervical Cancer. Clin. Transl. Oncol. 9: 347-354.

Morotomi-Yano, K., Akiyama, H., and Yano, K. 2011. Nanosecond Pulsed Electric Fields Activate MAPK Pathways in Human Cells. Biochem. Biophys. Res. Commun. 515: 99-106.

Morotomi-Yano, K., Oyadomari, S., Akiyama, H., and Yano, K. 2012. Nanosecond Pulsed Electric Act as A Novel Cellular Stress That Induces Translational Suppression Accompanied by elF2a Phosphorylation and 4E-BP1 Dephosphorylation. Nanosecond Pulsed Electric Fields Activate MAPK Pathways in Human Cells. Biochem. Biophys. Res. Commun. 318: 1733-1744.

Nagahama, M., Shimomura, N., Nakagawa, A., Teranishi, K., Uto, Y., and Hori, H. 2013. In Vivo Experimental Study of Nanosecond Pulsed Electric Field Effects on Solid Tumors. IEEE Transactions on Dielectrics and Electrical Insulation. 20: 1266-1272.

Nuccitelli, R., Chen, X., Pakhomov, A.G., Baldwin, W.H., Sheikh, S., Pomicter, J.L., Ren, W., Osgood, C., Swanson, R.J., Kolb, F.F., Beebe, S.J., and Schoenbach, K.H. 2009. A New Pulsed Electric Field Therapy for Melanoma Disrupts the Tumor’s Blood Supply and Causes Complete Remission without Recurrence. Int. J. Cancer. 125: 438-445.

Ren, W., and Beeb, S.J. 2011. An Apoptosis Targeted Stimulus With Nanosecond Pulsed Electric Fields (NsPEFs) in E4 Squamous Cell Carcinoma. Apoptosis. 16: 382-393.

Ren, Z., Chen, X., Cui, G., Yin, S., Chen, L., Jiang, J., Hu, Z., Xie, H., Zheng, S., and Zhou, L. 2013. Nanosecond Pulsed Electric Field Inhibits cancer Growth Followed by Alteration in Expression of NF-kB and Wnt/β-catenin Signaling Molecules. PLOSone. 8: 1-12.

Rohmah, M.K., Lyrawati, D., Yano, K. Nanosecond Pulsed Electric Fields (nsPEFs) Induce Socs1 and Socs3. Jurnal Kedokteran Brawijaya. Vol 28 (3): 1-7.

Singh, N., Hussain, S., Suri, V., Bharadwaj, M., Das, B. C., Sobti, R.C. 2011. Aberrant Expression of Trancription Factors STAT-3 and STAT-5 and Their Epigenetic Control by SOCS-1 Gene: The STAT Signaling Crostalk in HPV Coupled Cervical Carcinogenesis, Proc. AACR 102nd Annu. Meet. Doi: 10.1158/1538-7445.AM2011-2719.

Sobti RC, Singh N, Hussain S, Suri V, Bharti AC, and Das BC. Overexpression of STAT3 in HPV-mediated Cervical Cancer in a North Indian Population. Mololecular and Biochemistry. 2009; 330(1-2): 193-199.

Sobti RC, Singh N, Hussain S, Suri V, Bharadwaj M, and Das BC. Deregulation of STAT-5 Isoforms in the Development of HPV-Mediated Cervical Carcinogenesis. Journal of Receptor and Signal Transductions. 2010; 30(3): 178-188.

Stacey, M., Stickley, J. Fox, P., Statler, V., Schoenbach, K.H., Beebe, S.J., and Buescher, S. 2003. Differential Effects in Cells Exposed to Ultra-Short, High Intensity Electric Fields: Cell Survival, DNA Damage, and Cell Cycle Analysis. Mutation Res. 542: 65-75.

Wu, S., Wang, Y., Guo, J., Chen, Q., Zhang, J., and Fang, J. 2014. Nanosecond Pulsed Electric fields as a Novel Drug Free Therapy for Breast Cancer: An In Vivo Study. Cancer Letter. 343: 268-274.

Zou, H., Gan, X.L., Linghu, L.J., Chen, C., Hu, L.N., and Zhang, Y. 2013. Intense Nanosecond Pulsed Electric Fields Promote Cancer Cell Apoptosis Through Centrosome-Dependent Pathway Involving Reduced Level of PLK1. Eur. Rev. Med. Pharmacol. Sci. 17: 152-160.


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