Tiloke Charlette, Phulukdaree Alisa, Anand Krishnan, Gengan Robert M, Chuturgoon Anil A
Discipline of Medical Biochemistry and Chemical Pathology, School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal, Congella, Durban 4013, South Africa.
Department of Physiology, School of Medicine, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.
J Cell Biochem. 2016 Oct;117(10):2302-14. doi: 10.1002/jcb.25528. Epub 2016 Mar 11.
Gold nanoparticles (AuNP's) facilitate cancer cell recognition and can be manufactured by green synthesis using nutrient rich medicinal plants such as Moringa oleifera (MO). Targeting dysregulated oncogenes and tumor suppressor genes is crucial for cancer therapeutics. We investigated the antiproliferative effects of AuNP synthesized from MO aqueous leaf extracts (MLAuNP ) in A549 lung and SNO oesophageal cancer cells. A one-pot green synthesis technique was used to synthesise MLAuNP . A549, SNO cancer cells and normal peripheral blood mononuclear cells (PBMCs) were exposed to MLAuNP and CAuNP to evaluate cytotoxicity (MTT assay); apoptosis was measured by phosphatidylserine (PS) externalization, mitochondrial depolarization (ΔΨm) (flow cytometry), caspase-3/7, -9 activity, and ATP levels (luminometry). The mRNA expression of c-myc, p53, Skp2, Fbw7α, and caspase-9 splice variants was determined using qPCR, while relative protein expression of c-myc, p53, SRp30a, Bax, Bcl-2, Smac/DIABLO, Hsp70, and PARP-1 were determined by Western blotting. MLAuNP and CAuNP were not cytotoxic to PBMCs, whilst its pro-apoptotic properties were confirmed in A549 and SNO cells. MLAuNP significantly increased caspase activity in SNO cells while MLAuNP significantly increased PS externalization, ΔΨm, caspase-9, caspase-3/7 activities, and decreased ATP levels in A549 cells. Also, p53 mRNA and protein levels, SRp30a (P = 0.428), Bax, Smac/DIABLO and PARP-1 24 kDa fragment levels were significantly increased. Conversely, MLAuNP significantly decreased Bcl-2, Hsp70, Skp2, Fbw7α, c-myc mRNA, and protein levels and activated alternate splicing with caspase-9a splice variant being significantly increased. MLAuNP possesses antiproliferative properties and induced apoptosis in A549 cells by activating alternate splicing of caspase-9. J. Cell. Biochem. 117: 2302-2314, 2016. © 2016 Wiley Periodicals, Inc.
金纳米颗粒（AuNP's）有助于癌细胞识别，并且可以通过使用营养丰富的药用植物如辣木（MO）进行绿色合成来制造。靶向失调的癌基因和肿瘤抑制基因对于癌症治疗至关重要。我们研究了从MO叶水提取物（MLAuNP）合成的AuNP对A549肺癌细胞和SNO食管癌细胞的抗增殖作用。采用一锅法绿色合成技术合成MLAuNP。将A549、SNO癌细胞和正常外周血单个核细胞（PBMCs）暴露于MLAuNP和CAuNP中以评估细胞毒性（MTT法）；通过磷脂酰丝氨酸（PS）外化、线粒体去极化（ΔΨm）（流式细胞术）、caspase-3/7、-9活性和ATP水平（化学发光法）来测量细胞凋亡。使用qPCR测定c-myc、p53、Skp2、Fbw7α和caspase-9剪接变体的mRNA表达，同时通过蛋白质免疫印迹法测定c-myc、p53、SRp30a、Bax、Bcl-2、Smac/DIABLO、Hsp70和PARP-1的相对蛋白表达。MLAuNP和CAuNP对PBMCs无细胞毒性，而其促凋亡特性在A549和SNO细胞中得到证实。MLAuNP显著增加SNO细胞中的caspase活性，而MLAuNP显著增加A549细胞中的PS外化、ΔΨm、caspase-9、caspase-3/7活性并降低ATP水平。此外，p53的mRNA和蛋白水平、SRp30a（P = 0.428）、Bax、Smac/DIABLO和PARP-1 24 kDa片段水平显著增加。相反，MLAuNP显著降低Bcl-2、Hsp70、Skp2、Fbw7α、c-myc的mRNA和蛋白水平，并激活可变剪接，caspase-9a剪接变体显著增加。MLAuNP具有抗增殖特性，并通过激活caspase-9的可变剪接诱导A549细胞凋亡。《细胞生物化学杂志》117: 2302 - 2314, 2016。© 2016威利期刊公司。
