Podophyllotoxin Induces ROS-Mediated Apoptosis and Cell Cycle Arrest in Human Colorectal Cancer Cells via p38 MAPK Signaling

Abstract

Podophyllotoxin (PT), a lignan compound from the roots and rhizomes of Podophyllum peltatum, has diverse pharmacological activities including anticancer effect in several types of cancer. The molecular mechanism of the anticancer effects of PT on colorectal cancer cells has not been reported yet. In this study, we sought to evaluate the anticancer effect of PT on human colorectal cancer HCT116 cells and identify the detailed molecular mechanism. PT inhibited the growth of cells and colony formation in a concentration-dependent manner and induced apoptosis as determined by the annexin V/7-aminoactinomycin D double staining assay. PT-induced apoptosis was accompanied by cell cycle arrest in the G2/M phase and an increase in the generation of reactive oxygen species (ROS). The effects of PT on the induction of ROS and apoptosis were prevented by pretreatment with N-acetyl-L-cysteine (NAC), indicating that an increase in ROS generation mediates the apoptosis of HCT116 cells induced by PT. Furthermore, Western blot analysis showed that PT upregulated the level of phospho (p)-p38 mitogen-activated protein kinase (MAPK). The treatment of SB203580, a p38 inhibitor, strongly prevented the apoptosis induced by PT, suggesting that PT-induced apoptosis involved the p38 MAPK signaling pathway. In addition, PT induced the loss of mitochondrial membrane potential and multi-caspase activation. The results suggested that PT induced cell cycle arrest in the G2/M phase and apoptosis through the p38 MAPK signaling pathway by upregulating ROS in HCT116 cells.

Ononin Shows Anticancer Activity Against Laryngeal Cancer via the Inhibition of ERK/JNK/p38 Signaling Pathway

“…Regulating intracellular ROS status could proficiently kill tumor cells and suppress the adverse effects of radio/chemotherapy, and it is presently regarded as the primary means of tumor management (35). Several previous studies already demonstrated that many natural compounds are well known to increase the intracellular ROS production and lead to cell death in cancer cells (36)(37)(38). Likewise, we found that the ononin treatment substantially augmented the intracellular ROS status in the Hep-2 cells, thereby leading to oxidative stress mediated cell death (Figure 2B).…”

Biosynthesis, total synthesis, and pharmacological activities of aryltetralin-type lignan podophyllotoxin and its derivatives

“…In 2021, Lee evaluated the anticancer mechanism of PTOX on human colorectal cancer and found that it could induce cell cycle arrest in the G2/M phase and apoptosis through the p38 MAPK signalling pathway by upregulating reactive oxygen species (ROS) in HCT116 cells. 90 However, more applications of PTOX deal with its structural derivatives.…”

Traditional Chinese medicine for colorectal cancer treatment: potential targets and mechanisms of action

“… – AOM/DSS induced CRC model mice Proliferation ↓, inflammation ↓, Angiogenesis ↓, Invasion ↓ ↓: Ki-67, β-catenin, IL-1b, TNF-α, NF-κB, MMP9, Ereg, Muc16 NF-κB [ 74 ] Curcumin Jiang Huang ( Curcuma longa L.) HCT-116、SW620、HT-29 HCT-116 orthotopic mice Proliferation ↓, Invasion ↓, Angiogenesis ↓, Apoptosis ↑ ↓: NF-κB, COX2, Cyclin D1, c-myc, MMP9, ICAM-1, CXCR4, survivin, Bcl-2, VEGF NF-κB [ 75 ] Ginsenoside Rh1 (Rh1) Ren Shen ( Panax ginseng C.A.Mey.) SW620 – Proliferation ↓, Migration ↓, Invasion ↓ ↑: TIMP3 ↓: MMP1, MMP3, p-P38/P38, p-ERK1/2/ERK1-2, p-JNK/JNK MAPK [ 94 ] – SW620 xenograft mice Tumor growth ↓ ↓: p-P38/P38, p-ERK1/2/ERK1-2, p-JNK/JNK Diterpenoid C Jiang Huang ( Curcuma longa L.) SW620 – Proliferation ↓, Apoptosis ↑ ↑: caspase-3 ↓: p-ERK, p-JNK, p-p38 MAPK MAPK [ 96 ] Podophyllotoxin (PT) Gui Jiu ( Podophyllum peltatum ) HCT-116 – Apoptosis ↑, Block cell cycle ↑: ROS, GRP78, CHOP, p38 MAPK p38 MAPK [ 100 ] Emodin (EMD) Da Huang ( Rheum palmatum L.) SW620、SW480 – Apoptosis ↑, Migration ↓, Invasion ↓ …”

“…Podophyllotoxin (PT) is an active ingredient extracted from Podophyllum peltatum (Gui Jiu), which is highly cytotoxic to various cancer cells [97][98][99]. Lee et al [100] found that PT intervention increased the level of reactive oxygen species (ROS), upregulated the expression of ER stress markers GRP78 and CHOP, and increased the phosphorylation of p38 MAPK in HCT-116 cells, suggesting that PT could induce the p38 MAPK signaling pathway and ER stress-mediated apoptosis through upregulation of ROS in HCT-116 cells, accompanied by G2/M phase cell-cycle arrest (Fig. 3).…”