SAMRC/UP Precision Prevention and Novel Drug Targets for HIV-Associated Cancers Extramural Unit, Pan African Cancer Research Institute, Faculty of Health Sciences, University of Pretoria, Hatfield, 0028, South Africa; Department of Internal Medicine, School of Medicine, Faculty of Health Sciences, University of the Witwatersrand, Parktown, 2193, South Africa.
SAMRC/UP Precision Prevention and Novel Drug Targets for HIV-Associated Cancers Extramural Unit, Pan African Cancer Research Institute, Faculty of Health Sciences, University of Pretoria, Hatfield, 0028, South Africa.
Biomed Pharmacother. 2020 Dec;132:110829. doi: 10.1016/j.biopha.2020.110829. Epub 2020 Oct 12.
The Sub-Saharan countries, particularly South Africa has the largest number of people living with HIV, accompanied by the largest antiretroviral treatment (ART) programme in the world. The Highly Active Antiretroviral Treatment (HAART) is the most effective regimen against HIV/AIDS and has improved the lifespan and quality of life of HIV positive patients. HAART has also led to a decrease in the incidence of AIDS defining cancers (ADCs) while there is an increased incidence of the non-AIDS Defining Cancers (NADCs), such as lung cancer in the HAART era. The association between lung tumourigenesis and the use of HAART components such as the dual protease inhibitor (PI) lopinavir/ritonavir (LPV/r) is poorly understood. Using cell and molecular biological approaches, this study aimed at elucidating the effects of LPV/r on the regulation of the cell cycle related genes in normal (MRC-5) and adenocarcinoma (A549) lung cells. Initially, the nuclear integrity of these cells in response to LPV/r was determined using DAPI staining. The effect of LPV/r on cell cycle genes was evaluated through the use of a RT2 PCR gene array of 84 genes related to the cell cycle signaling pathway. The PCR array data was validated by Real-Time Quantification PCR (RT-qPCR). Ingenuity Pathway Analysis (IPA) bio-informatics tool was employed to disclose the molecular mechanism/s observed at cellular and gene expression levels. Loss of nuclear integrity and the upregulation of the p53 DNA damage response (DDR) pathway was revealed by DAPI staining, differential gene expression and IPA core analysis. Furthermore, MAD2L2 and AURKB which also play a role in the DDR pathway were shown to be differentially expressed. The activation of the CASP3 gene in response to LPV/r in A549 cells was also observed. The findings of this study suggest genotoxic properties of LPV/r in healthy normal lung fibroblasts cells and anti-tumour properties in the A549 cells.
撒哈拉以南非洲国家，特别是南非拥有世界上数量最多的艾滋病毒感染者，同时也拥有世界上最大的抗逆转录病毒治疗（ART）计划。高效抗逆转录病毒治疗（HAART）是对抗艾滋病毒/艾滋病最有效的方案，它提高了艾滋病毒阳性患者的寿命和生活质量。HAART 还降低了艾滋病定义性癌症（ADCs）的发病率，而非艾滋病定义性癌症（NADCs）的发病率则有所增加，例如在 HAART 时代，肺癌的发病率有所增加。HAART 与肿瘤发生和使用 HAART 成分（如双重蛋白酶抑制剂[PI]洛匹那韦/利托那韦[LPV/r]）之间的关联尚未得到充分理解。本研究使用细胞和分子生物学方法，旨在阐明 LPV/r 对正常（MRC-5）和肺腺癌（A549）细胞中细胞周期相关基因调控的影响。首先，通过 DAPI 染色确定这些细胞对 LPV/r 的核完整性。通过使用与细胞周期信号通路相关的 84 个基因的 RT2 PCR 基因芯片评估 LPV/r 对细胞周期基因的影响。PCR 芯片数据通过实时定量 PCR（RT-qPCR）进行验证。采用 IPA 生物信息学工具揭示细胞和基因表达水平观察到的分子机制/。DAPI 染色、差异基因表达和 IPA 核心分析显示，核完整性丧失和 p53 DNA 损伤反应（DDR）途径的上调。此外，还显示 MAD2L2 和 AURKB 这两个也在 DDR 途径中起作用的基因表达差异。还观察到 LPV/r 对 A549 细胞中 CASP3 基因的激活。本研究的结果表明 LPV/r 在健康正常肺成纤维细胞中的遗传毒性特性以及在 A549 细胞中的抗肿瘤特性。
