Rachmawati Heni, Jessica Adhitya, Sumirtaputra Yeyet Cahyati, Retnoningrum Debbie Sofie, Adlia Amirah, Ningrum Ratih Asmana
School of Pharmacy, Bandung Institute of Technology, Ganesha 10, 40132, Bandung, Indonesia.
Department of Pharmacokinetics and Drug Delivery, University of Groningen, Antonius Deusinglaan I, Groningen, Netherlands.
Sci Pharm. 2016 Feb 14;84(1):113-30. doi: 10.3797/scipharm.ISP.2015.04. Print 2016 Jan-Mar.
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 (rhIFNα2b) by using a synthetic gene. In addition, two mutein forms of rhIFNα2b 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 rIFNα2b 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 rhIFNα2b. There was no significant difference in the MTT assay and cell viability after cells were treated with both forms of rhIFNα2b. However, the mutein rhIFNα2b tended to show better proapoptotic activity reflected by flow cytometric data, protein expression of pSTAT1, and DNA expression of caspase 3.
干扰素α2b是治疗肝炎病毒感染的唯一标准治疗性蛋白质。进一步研究表明，这种蛋白质在几个癌性器官中也具有抗肿瘤活性。这种抗肿瘤活性的一个主要途径是通过抗增殖以及促凋亡作用介导的。此前，我们通过使用合成基因成功开发了重组人干扰素α2b（rhIFNα2b）。此外，还产生了两种rhIFNα2b的突变体形式以改善这种蛋白质的特性。与单点突变以及天然形式相比，两点突变显示出更好的药代动力学特征。在本研究中，使用HepG2细胞在体外研究了这种突变体形式的抗肿瘤作用。作为比较，使用了天然形式以及市售的rIFNα2b。研究了几个参数，包括MTT法、细胞活力测试、使用流式细胞术分析的细胞周期以及参与细胞生长的基因和蛋白质表达。观察后者以研究rhIFNα2b的作用机制。用两种形式的rhIFNα2b处理细胞后，MTT法和细胞活力没有显著差异。然而，突变体rhIFNα2b倾向于表现出更好的促凋亡活性，这由流式细胞术数据显示，pSTAT1的蛋白质表达和caspase 3的DNA表达所反映。
