Effects of 5,6-Dihydroxy-2,4-Dimethoxy-9,10-Dihydrophenanthrene on G₂/M Cell Cycle Arrest and Apoptosis in Human Lung Carcinoma Cells

Abstract

5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (HMP) is an active compound isolated from the rhizome extracts of Dioscorea membranacea Pierre, a Thai medicinal plant. This study aimed to investigate the growth-inhibitory and apoptosis-inducing effects of HMP in human lung cancer A549 cells. The antiproliferative and cytotoxic effects of HMP were analyzed by a Sulforhodamine B assay. Cell division, cell cycle distribution and membrane asymmetry changes were each performed with different fluorescent dyes and then analyzed by flow cytometry. Real-time PCR and immunoblotting were used to detect cell cycle- and apoptosis-related mRNA levels and proteins, respectively. The nuclear morphology of the cells stained with DAPI and DNA fragmentation were detected by fluorescence microscopy and gel electrophoresis, respectively. The results showed that HMP exerted strong antiproliferative and cytotoxic activities in A549 cells with the highest selectivity index. It halted the cell cycle in [Formula: see text]/M phase via down-regulation of the expression levels of regulatory proteins Cdc25C, Cdk1 and cyclinB1. In addition, HMP induced early apoptotic cells with externalized phosphatidylserine and subsequent apoptotic cells in sub-[Formula: see text] phase. HMP increased caspase-3 activity and levels of the cleaved (active) form of caspase-3 whose actions were supported by the cleavage of its target PARP, nuclear condensation and DNA apoptotic ladder. Moreover, HMP significantly increased the mRNA and protein levels of proapoptotic Bax as well as promoted subsequent caspase-9 activation and BID cleavage, indicating HMP-induced apoptosis via both intrinsic and extrinsic pathways. These data support, for the first time, the potential role of HMP as a cell-cycle arrest and apoptosis-inducing agent for lung cancer treatment.

Effects of Wild Yam Root (Dioscorea villosa) Extract on the Gene Expression Profile of Triple-negative Breast Cancer Cells

“…These effects were concurrent with single gene up-regulation of the p21 gene CDKN1A, all of which are likely responsible for the observed cytostatic effects of WYE in a 6-day proliferation study. Our findings in this aspect support much of the existing literature for WYE and saponins from the Dioscorea species having the capacity to induce cell-cycle arrest (G 2 /M) across diverse cancer cell lines with observed downregulation of a similar list of cyclins and cell cycle regulatory transcripts (Cdc25C, Cdk1) (31)(32)(33)(34).…”

A Combined Phytochemistry and Network Pharmacology Approach to Reveal Potential Anti-NSCLC Effective Substances and Mechanisms in Marsdenia tenacissima (Roxb.) Moon (Stem)

“…Generally, various protein molecules participate in regulating the mitochondrial apoptotic pathway such as pro-apoptotic members (Bax and Bad) and anti-apoptotic members (Bcl-2 and Bcl-xl) ( Dong et al, 2019 ). Moreover, the pathway activated specific pivotal proteinases such as initiator caspase-9 and effector caspase-3 and subsequently resulted in DNA fragmentation as well as nuclear PARP degradation during apoptosis ( Duangprompo et al, 2016 ; Chao et al, 2018 ). Therefore, Annexin V-FITC/PI staining was conducted to detect apoptotic alterations that may be related to A549 cells cytotoxicity.…”