Antiproliferative Activity of Recombinant Human Interferon Alpha2b on Estrogen Positive Human Breast Cancer McF-7 Cell Line

Ningrum, Ratih Asmana; Wisnuwardhani, Popi Hadi; Santoso, Adi; Herawati, Neng

doi:10.14499/indonesianjpharm26iss2pp86

Cited by 8 publications

(15 citation statements)

References 29 publications

(23 reference statements)

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“…The huIFNα2b dosage is more than 10‐fold lower than the dosages reported for huIFNα2b production by E. coli and P. pastoris X33 cell lines to achieve comparable drug efficacy (Ningrum et al . , ). This antiproliferative effect is a result of both apoptosis and cell cycle arrest (Roos et al .…”

Section: Discussionmentioning

confidence: 99%

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

et al. 2019

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Aims The present study was aimed at design of experiments (DoE)‐ and artificial intelligence‐based culture medium optimization for high level extracellular production of a novel recombinant human interferon alpha 2b (huIFNα2b) in glycoengineered Pichia pastoris and its characterization. Methods and Results The artificial neural network—genetic algorithm model exhibited improved huIFNα2b production and better predictability compared to response surface methodology. The optimized medium exhibited a fivefold increase in huIFNα2b titre compared to the complex medium. A maximum titre of huIFNα2b (436 mg l−1) was achieved using the optimized medium in the bioreactor. Real‐time capacitance data from dielectric spectroscopy were utilized to model the growth kinetics with unstructured models. Biological characterization by antiproliferative assay proved that the purified recombinant huIFNα2b was biologically active, exhibiting growth inhibition on breast cancer cell line. Conclusions Culture medium optimization resulted in enhanced production of huIFNα2b in glycoengineered P. pastoris at both shake flask and bioreactor level. The purified huIFNα2b was found to be N‐glycosylated and biologically active. Significance and Impact of the Study DoE‐based medium optimization strategy significantly improved huIFNα2b production. The antiproliferative activity of huIFNα2b substantiates its potential scope for application in cancer therapy.

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Section: Discussionmentioning

confidence: 99%

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

et al. 2019

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“…Further treatment was applied by various concentration of hIFNα-2a for 5 days. The treatment conditions were based on a report by Ningrum et al (2015).…”

Section: Antiproliferative Assay Cell Cultification and Treatmentsmentioning

confidence: 99%

“…The combination of NAT-hIFNα-2a and tamoxifen is reported to be very effective in inhibiting proliferation of estrogen positive MCF-7 cell line (Seymour and Bezweda 1993;Ningrum et al 2015;Wathon et al 2015). In this study we combined purified NAT-hIFNα-2a with various concentrations (2, 4, and 8 µg/mL) with tamoxifen (1µM) to investigate the effect of its antiproliferative activity by MTT colorimetric assay.…”

Section: Anti-proliferative Activitymentioning

confidence: 99%

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Overproduction, characterization and preliminary antiproliferative activity determination of non-tagged recombinant human interferon alpha-2a produced in Pichia pastoris

2017

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Abstract. Herawati N, Wardiana A, Ningrum RA. 2017. Overproduction, characterization and antiproliferative activity determination of non-tagged recombinant human interferon alpha-2a produced in Pichia pastoris. Nusantara Bioscience 9: 97-101. Recombinant human interferon alpha-2a (rhIFNα-2a) has been widely used for antiviral, anticancer as well as immunomodulatory clinical therapy. In our previous research, we constructed the Open Reading Frame (ORF) encoding synthetic rhIFNα-2a to be in framed with N-terminal alpha factor secretion system in pPICZαB shuttle vector. This research covered overproduction, characterization of the protein and preliminary determination of biological potency assay by using the estrogen positive cell line MCF-7. To overproduce the protein, cultivation was performed in Buffered complex medium containing glycerol (BMGY) for 24 h and production was performed in Buffered complex medium containing methanol (BMMY) during 48 hours at 30°C. The recombinant protein was purified by affinity chromatography using blue sepharose resin. Analysis of amino acid sequence by using MS/MS2 mass spectrometry covered 21% of the total amino acid residues. To determine the initial biological activity, the effect of rhIFNα-2a on MCF-7 breast cancer cell line was studied in vitro. The anti-proliferative activity of hIFNα-2a was performed by 3-[4.5-dimethyltiazol-2il]-2.5 diphenylltetrazolium bromide (MTT) assay. The result showed that the protein is able to inhibit MCF-7 proliferation and the reduction of cell growth was dose-dependently. To monitor the protein production, we performed stability expression assay as well. The result informed that the expression of ORF still stable until 60 th generation

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“…The use of rhIFNα2a as recombinant protein therapeutic has been approved by the United States Food and Drug Administration (US FDA) since 1986. [5][6][7] This mini review is focused on the use of hIFNα2a as therapeutic protein against hepatitis B and C.…”

Section: Introductionmentioning

confidence: 99%

Human Interferon Alpha2a as Anti Hepatitis B and C

Ningrum

2017

Indones J Clin Pharm

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Hepatitis is an inflammation of the liver mainly caused by hepatitis viruses. There are 5 different types of hepatitis based on the infecting virus; A, B, C, D and E. Hepatitis B and C are chronic diseases that potentially develop into hepatocarcinoma and cirrhosis on unappropriate treatments. World Health Organization (WHO) stated that currently 350 million people worldwide are living with chronic hepatitis B and 150 million people are living with Hepatitis C. The mortality rate in the world due to hepatitis is about 1.5 million people per year. The human interferon alpha2a (hIFNα2a) is a therapeutic protein used as therapeutic protein for hepatitis B and C. This review discusses the hepatitis B (HBV) and C (HCV) viruses, mechanisms of hIFNα2a as antivirus through signal transduction pathway and improvement of hIFNα2a properties by protein modification. The application of recombinant hIFNα2a (rhIFNα2a) in the treatment of hepatitis B and C that recommended by European Association for The Study of Liver (EASL) and the viral resistance mechanism are also included. The status of hepatitis B and C and the development of rhIFNα2a is also described as well.Keywords: Antiviral, hepatitis, human interferon alpha2a, protein modification, viral resistance Protein Interferon Alfa-2a Manusia sebagai Anti Hepatitis B dan C AbstrakHepatitis merupakan kondisi inflamasi pada hati yang terutama disebabkan oleh virus hepatitis. Berdasarkan tipe virus yang menginfeksi, terdapat lima jenis penyakit hepatitis yaitu A, B, C, D dan E. Hepatitis B dan C merupakan penyakit kronis yang berpotensi menjadi kanker hati dan sirosis jika tidak ditangani dengan baik. World Health Organization (WHO) menyatakan bahwa saat ini terdapat 350 juta orang yang terinfeksi hepatitis B dan 150 juta orang terinfeksi hepatitis C di seluruh dunia. Angka kematian yang disebabkan hepatitis mencapai 1,5 juta orang per tahun. Protein interferon alfa-2a manusia (hIFNα2a) adalah protein terapeutik yang digunakan sebagai obat hepatitis B dan C. Review ini mendiskusikan mengenai virus hepatitis B (HBV) dan C (HCV), mekanisme hIFNα2a sebagai antivirus melalui sistem transduksi sinyal dan peningkatan sifat hIFNα2a melalui modifikasi protein. Review ini juga membahas aplikasi bentuk rekombinan hIFNα2a (rhIFNα2a) dalam penanganan hepatitis B dan C yang direkomendasikan oleh European Association for The Study of Liver (EASL) dan mekanisme resistensi virus. Status hepatitis B dan C serta perkembangan rhIFNα2a juga didiskusikan lebih lanjut.

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Antiproliferative Activity of Recombinant Human Interferon Alpha2b on Estrogen Positive Human Breast Cancer McF-7 Cell Line

Cited by 8 publications

References 29 publications

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

High level extracellular production of recombinant human interferon alpha 2b in glycoengineeredPichia pastoris:culture medium optimization, high cell density cultivation and biological characterization

Overproduction, characterization and preliminary antiproliferative activity determination of non-tagged recombinant human interferon alpha-2a produced in Pichia pastoris

Human Interferon Alpha2a as Anti Hepatitis B and C

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