Removing Cystein Group On Interferon Alpha 2b at Position 2 and 99 does Not Diminish Antitumor Activity of the Protein, Even Better

Heni Rachmawati*, Adhitya Jessica, Yeyet Cahyati Sumirtaputra, Debbie Sofie Retnoningrum, Amirah Adlia, Ratih Asmana Ningrum

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Interferon alpha 2b is the only standard therapeutic protein for hepatitis virus infections. Further study demonstrated that this protein also posseses antitumor activity in several cancerous organs. One main pathway of this antitumor activity is mediated through antiproliferation as well as proapoptotic effects. Previously, we have successfully developed recombinant human interferon alpha 2b (rhIFN alpha 2b) by using a synthetic gene. In addition, two mutein forms of rhIFN alpha 2b were generated to improve the characteristics of this protein. Two point mutations showed better pharmacokinetic profiles than one point mutation as well as the native form. In the present study, this mutein form was studied for ist antitumor effect in vitro using HepG2 cells. As a comparison, the native form as well as a commercial rIFN alpha 2b were used. Several parameters were investigated including the MTT assay, cell viability test, cell cycle using flow cytometric analysis, and the genes and protein expressions involved in cell growth. The latest was observed to study the mechanism of rhIFN alpha 2b. There was no significant difference in the MTT assay and cell viability after cells were treated with both forms of rhIFN alpha 2b. However, the mutein rhIFN alpha 2b tended to show better proapoptotic activity reflected by flow cytometric data, protein expression of pSTAT1, and DNA expression of caspase 3.

Original languageEnglish
Pages (from-to)113-130
Number of pages18
JournalScientia Pharmaceutica
Volume84
Issue number1
DOIs
Publication statusPublished - 2016

Keywords

  • Interferon alpha 2b
  • Antiproliferation
  • Apoptosis
  • p21k1
  • p27
  • Caspase 3
  • pSTAT1
  • Flow cytometri
  • Mutein
  • CELL-LINE
  • GROWTH-REGULATION
  • LEUKEMIC-CELLS
  • ALPHA
  • ARREST
  • ACTIVATION
  • APOPTOSIS
  • KINASES
  • EXPRESSION
  • INHIBITORS

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