Original Article Effects of HIV protease, nucleoside/non-nucleoside reverse transcriptase inhibitors on Bax, Bcl-2 and apoptosis in two cervical cell lines Gbenga Anthony Adefolaju a, *, Kathrine Elizabeth Theron b , Margot Jill Hosie a a School of Anatomical Sciences, Wits Medical School, University of the Witwatersrand, 7, York Road, Parktown, 2193 Johannesburg, South Africa b Division of Human Genetics, Department of Medical Sciences, University of Limpopo, Private Bag x1106n, Sovenga 0727, South Africa 1. Introduction Highly active antiretroviral therapy (HAART), a multidrug combination regimen commonly consisting of nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcrip- tase inhibitors (NNRTIs) and protease inhibitors (PIs), has dramatically reduced mortality and morbidity among human immunodeficiency virus-1 (HIV-1) infected individuals [1,2]. However, continuous administration of the drugs in the cocktail has been associated with multiple adverse effects [2]. The focus of the initial development of the antiretroviral drugs was on clinical efficacy (reduction of mortality) – before all other considerations [3]. However, as AIDS has been better managed, emphasis on the long-term adverse effects induced by this therapy has also increased [3]. For example, while NRTI drug combinations provide enhanced anti-HIV therapy/prophylaxis, individual NRTIs, are implicated as possible chemical carcinogens and mutagens [4–7]. Studies on the effects of HAART on the incidence and progression of HIV/AIDS-associated cancers; non-Hodgkin lymphoma, cervical cancer and Kaposi’s sarcoma have provided contrasting data [8]. While there has been a decrease in the incidence of Kaposi’s sarcoma, HAART has reportedly not had a significant impact on the incidence of the other two AIDS defining malignancies [8] – some evidence even suggests an increase in these cancers [9]. Also, it has been widely reported [8–12] that the widespread use of HAART has increased the risk of non-AIDS defining malignancies (NADMs). In a review of the potential causes of the increased incidence of NADMs in the HAART era and also, as a result of HAART use, Nguyen et al. [11] lists as key factors; oncogenic viruses, (Epstein– Barr virus, Human papillomavirus, hepatitis-B, hepatitis-C), impaired immune surveillance, direct HIV-1 oncogenic effect, longer life expectancy associated with HAART (providing the longer latency period necessary for the development of certain cancers) and other traditional risk factors, such as smoking, sun exposure, alcohol, illicit drug use and ethnicity. Whether the antiretroviral compounds themselves are oncogenic is not clear [12]. Powles et al. [12] reported that there may be an association between the use of NNRTIs and the development of non-AIDS defining malignancies. Reports from in vitro studies suggest that antiretroviral drugs could either cause or potentially prevent the development of cancer [13,14]. This information, along with the evidence that Biomedicine & Pharmacotherapy 68 (2014) 241–251 A R T I C L E I N F O Article history: Received 24 July 2013 Accepted 10 August 2013 Keywords: Antiretroviral drugs Bax/Bcl-2 Apoptosis A B S T R A C T Protease inhibitors (PIs) and reverse transcriptase drugs are important components of highly active antiretroviral therapy (HAART) for treating human acquired immunodeficiency syndrome (AIDS). Long- term clinical therapeutic efficacy and treatment compliance of these agents have been limited by undesirable adverse effects and their oncogenicity has been queried. This study investigated the effects of selected antiretroviral agents on the expression of key apoptotic regulatory genes; Bax and Bcl-2 in two cervical cell lines HCS-2 and NCE16IIA by real-time qPCR gene expression and immunocytochemistry. The anti-apoptotic effects of the PI-LPV/r were investigated by cell death detection ELISA and acridine orange staining. All the antiretroviral drugs and combinations tested had no effects on Bax and Bcl-2 gene expression and protein localisation in both cell lines. The protease inhibitors–LPV/r exhibited significant (P < 0.05) inhibition of camptothecin-induced apoptosis in the cervical cancer HCS-2 cell line but not in the normal immortalised NCE16IIA cell line. This anti-apoptotic property of HIV protease inhibitors, although shown so far not to involve protein and RNA synthesis might promote the development of cancer. ß 2013 Elsevier Masson SAS. All rights reserved. * Corresponding author. E-mail addresses: gbenga.adefolaju@wits.ac.za, adefolajugbenga@gmail.com (G.A. Adefolaju). Available online at www.sciencedirect.com 0753-3322/$ – see front matter ß 2013 Elsevier Masson SAS. All rights reserved. http://dx.doi.org/10.1016/j.biopha.2013.08.007