Gene expression analysis of apoptosis pathway in HeLa S3 cells subjected to nanosecond pulsed electric fields

Abstract Nanosecond pulsed electric fields (nsPEFs) have been extensively studied with respect to cellular responses. Whether nsPEFs can regulate gene expression and to modulate the synthesis of valuable compounds, has so far been only tested in the context of apoptosis in cancer cells. We used the unicellular algae Haematococcus pluvialis as system to test, whether nsPEFs could alter gene expression and to promote the biosynthesis of astaxanthin. We find that nsPEFs induce a mild, but significant increase of mortality up to about 20%, accompanied by a moderate increase of astaxanthin accumulation. Steady-state transcript levels of three key genes psy, crtR-b and bkt 1 were seen to increase with a maximum at 3 d after PEF treatment at 50 ns. Pulsing at 25 ns reduce the transcripts of psy, crtR-b from around day 2 after the pulse, while those of bkt 1 remain unchanged. By blocking the membrane-located NADPH oxidase RboH, diphenylene iodonium by itself increased both, the levels of astaxanthin and transcripts of all three biosynthetic genes, and this increase was added up to that produced by nsPEFs. Artificial calcium influx by an ionophore did not induce major changes in the accumulation of astaxanthin, nor in the transcript levels, but amplified the response of crtR-b to nsPEFs at 25 ns, while decreased in 50 ns treatment. When Ca2+ influx was inhibited by GdCl3, the transcript of psy and bkt 1 were decreased for both 25 ns and 50 ns treatments, while crtR-b exhibited an obvious increase for the 25 ns treatment. We interpret these data in a working model, where nsPEFs permeabilise plasma and chloroplast membrane depending on pulse duration leading to a differential release of plastid retrograde signaling to the nucleus.

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Study of effect of pulsing sequence of nanosecond pulsed electric fields on viability of HeLa S3 cell

2010 IEEE International Power Modulator and High Voltage Conference ◽

10.1109/ipmhvc.2010.5958329 ◽

2010 ◽

Cited By ~ 2

Author(s):

Kazunori Mitsutake ◽

Akimitsu Satoh ◽

Shingo Mine ◽

Keisuke Abe ◽

Sunao Katsuki ◽

...

Keyword(s):

Electric Fields ◽

Pulsed Electric Fields ◽

Hela S3 ◽

Nanosecond Pulsed Electric Fields

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Nanosecond pulsed electric fields enhance mesenchymal stem cells differentiation via DNMT1-regulated OCT4/NANOG gene expression

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01821-5 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Kejia Li ◽

Tong Ning ◽

Hao Wang ◽

Yangzi Jiang ◽

Jue Zhang ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Electric Fields ◽

Pulsed Electric Fields ◽

Nanog Gene ◽

Nanosecond Pulsed Electric Fields

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Pengaruh Variasi Jumlah Tembakan Nanosecond Pulsed Electric Fields (Nspefs) Terhadap Ekspresi Gen Socs3 pada Sel Kanker Serviks Hela S3

JRST (Jurnal Riset dan Sain Teknologi) ◽

10.30595/jrst.v1i2.1702 ◽

2017 ◽

Vol 1 (2) ◽

pp. 45

Author(s):

Martina Kurnia Rohmah

Keyword(s):

Gene Expression ◽

Real Time ◽

Electric Fields ◽

Real Time Pcr ◽

Pulsed Electric Fields ◽

Rt Pcr ◽

Kolmogorov Smirnov ◽

Hela S3 ◽

Short Time ◽

Research Show

AbstrakNanosecond 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 AbstractNanosecond 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

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