Pengaruh Variasi Jumlah Tembakan Nanosecond Pulsed Electric Fields (Nspefs) Terhadap Ekspresi Gen Socs3 pada Sel Kanker Serviks Hela S3
DOI:
https://doi.org/10.30595/jrst.v1i2.1702Keywords:
NsPEFs, shot, expression, Socs3Abstract
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.
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Kata Kunci: NsPEFs, tembakan, ekspresi, Socs3
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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.
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Keywords: NsPEFs, shot, expression, Socs3
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