Novel Antitumor Invasive Actions of <i>p</i>-Cymene by Decreasing MMP-9/TIMP-1 Expression Ratio in Human Fibrosarcoma HT-1080 Cells

Abstract

p-Cymene (4-isopropyltoluene) has been reported to have beneficial actions such as anti-inflammatory and antinociceptive activities. To evaluate whether p-cymene exhibits antitumor invasive actions, we examined the effects of p-cymene on the production of matrix metalloproteinase 9 (MMP-9)/gelatinase B and tissue inhibitor of metalloproteinases-1 (TIMP-1) in human fibrosarcoma HT-1080 cells. p-Cymene was found to dose-dependently inhibit the 12-O-tetradecanoylphorbol 13-acetate (TPA)-augmented production and gene expression of MMP-9 in HT-1080 cells. In contrast, p-cymene enhanced the TPA-augmented production and gene expression of TIMP-1 in HT-1080 cells. However, there was no change in the constitutive level of MMP-9 and TIMP-1 mRNAs and TIMP-1 protein in p-cymene-treated cells. In addition, we found that the in-vitro TPA-augmented invasiveness of HT-1080 cells was inhibited by p-cymene in a dose-dependent manner. Furthermore, p-cymene was found to suppress the constitutive and/or TPA-augmented phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and p38 mitogen-activated protein kinase (MAPK) in HT-1080 cells. Thus, these results provide novel evidence that p-cymene is an effective candidate for the prevention of tumor invasion and metastasis through mechanisms that include the inhibition of MMP-9 expression and the augmentation of TIMP-1 production along with the suppression of ERK1/2 and p38 MAPK signal pathways in tumor cells.Key words p-cymene; tumor invasion; matrix metalloproteinase 9; tissue inhibitor of metalloproteinases-1; extracellular signal-regulated kinase 1/2 (ERK1/2); p38 mitogen-activated protein kinase (p38 MAPK)The proteolytic degradation of extracellular matrix (ECM) components in basement membranes and stroma tissues is requisite for progressing the invasion and metastasis of malignant tumor cells. Matrix metalloproteinases (MMPs) play a crucial role in ECM degradation, and different sets of MMPs concertedly act in ECM breakdown. 1,2) Gelatinase A (72-kDa type IV collagenase)/MMP-2 and gelatinase B (92-kDa type IV collagenase)/MMP-9 have been reported to facilitate tumor cell invasion and epithelial-mesenchymal transition as a promoter and/or modulator in normal and tumor cells in vivo and in vitro.3) On the other hand, the enzymic activity of MMPs and proteolytic activation of the MMP precursor (proMMP) have been reported to be controlled by endogenous inhibitors such as tissue inhibitors of metalloproteinases (TIMPs).4) Since four TIMPs have been identified, designated as TIMP-1, TIMP-2, TIMP-3, and TIMP-4, they inhibit MMPs by forming a strong noncovalent complex with a 1 : 1 stoichiometry. 4) Regarding the inhibitory properties among the TIMPs, TIMPs can bind to different sets of MMPs: e.g., the preference inhibition of TIMP-1 against MMP-9, MMP-3, and MMP-1, and that of TIMP-2 against MMP-2 rather than MMP-9.4,5) Therefore, the inhibition of expression and/or enzymic activity of MMP is likely to be an effective target(s) for preventing tumor invasion and metastasis.

Update on Monoterpenes as Antimicrobial Agents: A Particular Focus on p-Cymene

“…The anti-inflammatory activity of p-cymene at least partly justifies its antinociceptive activity, as recorded in several in vivo studies performed on murine model systems [ 8 , 10 , 11 ]. Moreover, recent investigations report the antitumor activity of p-cymene and metal p-cymene complexes such as rutenium (II)- and osmium (II)-p-cymene complexes, which inhibit tumor proliferation through an antiangiogenic mechanism, cancer cell cytotoxicity, and anti-adhesion activity [ 12 , 13 , 14 ]. p-Cymene is the main antimicrobial compound in thyme, and a large body of evidence suggests that this monoterpene possesses antibacterial, antiviral and antifungal activities.…”

Comprehensive Analysis of the Chemical Composition and In Vitro Cytotoxic Mechanisms of Pallines Spinosa Flower and Leaf Essential Oils Against Breast Cancer Cells

“…The differences between the IC 50 values of F-PSEO and L-PSEO are attributed to both compositional and quantitative variations among these oils. The cyclic monoterpene p -cymene, that constitutes a 27.76% (by SPME, 1.33% by hydrodistillation) of the flower oil compared to 5.56% (SPME, 0.1%-hydrodistillation) in L-PSEO, has been reported to exhibit strong antitumor invasive and metastatic actions towards the human fibrosarcoma HT-1080 cells through inhibition of matrix metalloproteinase 9 (MMP-9) along with the suppression of ERK1/2 and p38 MAPK [50]. The cyclic monoterpene α-phellandrene, comprises 13.59% (SPME, 1.74%-hydrodistillation, compared to 1.08% and 0.36% in L-PSEO, respectively), induces apoptosis through the mitochondria-dependent pathway, cell cycle arrest at G0/G1 phase and production of reactive oxygen species (ROS) in murine WEHI-3 leukemic cells [51].…”

“…This is particularly obvious with the finding that the general caspase-inhibitor is only able to reverse ∼30% of the oil induced apoptosis in MDA-MB-2 31 compared to ∼60% in MCF-7 cells. The fact that the essential oils of P. spinosa contain compounds that promote cell death by autophagy [53], activation of MAPKs [50], production of ROS [51] and cell cycle arrest [51], proposes that they may also contribute to the cytotoxicity of F-PSEO and L-PSEO.…”

p-Cymene attenuates cancer pain via inhibitory pathways and modulation of calcium currents

“…The movement-evoked pain test did not show significant inhibition in the reduction of paw utilization by the mice (supplementary material). However, another study (Li et al, 2016) demonstrated that PC has the antitumor effect due to the modulation of ERK1/2 and p38 MAPK signal pathways in tumor cells.…”