cytochrome-c-t and Esophageal-Squamous-Cell-Carcinoma

Along with changes in dietary habits and lifestyle, the incidence of esophageal cancer is increasing around the world. Since chemotherapy for esophageal cancer has significant side effects, phytochemicals have attracted attention as an alternative medicine. Licochalcone C (LCC) is a flavonoid compound extracted from Licorice, with a variety of clinical uses including anti-cancer, anti-inflammatory and anti-oxidant effects. Treatment with LCC for 48 h significantly decreased cell viability of esophageal squamous cell carcinoma (ESCC) cells in a dose- and time-dependent manner with IC

Topics: Antineoplastic Agents; Apoptosis; Caspases; Cell Line, Tumor; Cell Proliferation; Cell Survival; Chalcones; Cytochromes c; Dose-Response Relationship, Drug; Endoplasmic Reticulum Chaperone BiP; Esophageal Squamous Cell Carcinoma; G1 Phase; Humans; Membrane Potential, Mitochondrial; Reactive Oxygen Species; Time Factors

Esophageal cancer is the eighth most common cancer globally. Transforming growth factor β regulator 4 (TBRG4) and caveolin-1 (CAV-1) are implicated in tumor progression. The aim of this study was to investigate the expressions of TBRG4 and CAV-1 in esophageal squamous cell carcinoma (ESCC), and their relationship with reactive oxygen species (ROS) formation. Human ESCC cell lines (EC9706, TE-1, and Eca109), and normal esophageal mucosal cell line (Het-1) were used in this study. The silencing of TBRG4 and/or CAV-1 by sh-RNA or overexpression of CAV-1 after TBRG4 knockdown was used to assess ROS levels. The results showed that down-regulation of TBRG4 reduced CAV-1 expression, and promoted ROS formation in ESCCs (p < 0.01). However, CAV-1 overexpression increased the expression level of TBRG4, but decreased ROS level in EC9706 cells (p < 0.01). Similarly, TBRG4 knockdown significantly reduced CAV-1 expression, promoted ROS formation, and caused cell cycle arrest at G0/G1 phase (p < 0.01). Caveolin-1 (CAV-1) knockdown also promoted cell apoptosis, cellular ROS formation and cell cycle arrest at G0/G1 phase (p < 0.01). However, CAV-1 overexpression in sh-TBRG4-treated EC9706 cells significantly upregulated TBRG4 expression, but significantly reduced the level of ROS, and inhibited cell-cycle arrest and apoptosis (p < 0.01). The enhancements in bcl-2/bax ratio, cytochrome c expression, and ROS levels by sh-TBRG4 were significantly reversed by CAV-1 overexpression in EC9706 cells. These results show that the upregulated expression of TBRG4 or CAV-1 promotes ESCC progression via regulation of intracellular ROS levels and inhibition of mitochondria-dependent apoptotic pathway.

Topics: Apoptosis; bcl-2-Associated X Protein; Caveolin 1; Cell Cycle; Cell Line, Tumor; Cytochromes c; Disease Progression; Esophageal Squamous Cell Carcinoma; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Gene Silencing; Humans; Mitochondrial Proteins; Reactive Oxygen Species; RNA-Binding Proteins; RNA, Small Interfering

α-Hederin has been shown promising anti-tumor potential against various cancer cell lines. However, reports about effects of α-hederin on esophageal squamous cell carcinoma (ESCC) are still unavailable.. To investigate the inhibitory effects of α-hederin on ESCC and explore the underlying mechanism.. Human esophageal carcinoma cell line (Eca-109) was used for the experiment. Cell Counting Kit-8, flow cytometry, Hoechst 33258 staining, enhanced ATP assay kit, 2',7'-dichlorofluorescin diacetate, JC-1 kit, and Western bolt were used to assess the cell viability, cycle, apoptosis, cellular ATP content, reactive oxygen species (ROS) level, mitochondrial membrane potential (MMP), and protein expression, respectively, in vitro. Xenografted tumor model was constructed to evaluate the in vivo anti-tumor effects of α-hederin.. Compared with control group, α-hederin significantly inhibited the proliferation, induced apoptosis of ESCC, and arrested the cell cycle in G1 phase (P < 0.05). α-Hederin induced the accumulation of ROS, decrement of ATP levels, and disruption of MMP (P < 0.05). The detection of mitochondrial and cytosol proteins showed that AIF, Apaf-1, and Cyt C were released and increased in cytoplasm, and then, caspase-3, caspase-9, and Bax were involved and increased, while Bcl-2 level was decreased (P < 0.05). Furthermore, the above changes were amplified in the group pretreated with L-buthionine sulfoximine, while N-acetyl-L-cysteine plays an opposite role (P < 0.05). Meanwhile, α-hederin significantly inhibited the growth of xenografted tumors with favorable safety.. α-Hederin could inhibit the proliferation and induce apoptosis of ESCC via dissipation of the MMP with simultaneous ROS generation and activation of the mitochondrial pathway.

Topics: Adenosine Triphosphate; Animals; Apoptosis; Apoptosis Inducing Factor; Apoptotic Protease-Activating Factor 1; bcl-2-Associated X Protein; Caspase 3; Caspase 9; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cytochromes c; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Flow Cytometry; Humans; In Situ Nick-End Labeling; In Vitro Techniques; Male; Membrane Potential, Mitochondrial; Mice, Nude; Mitochondria; Neoplasm Transplantation; Oleanolic Acid; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species; Saponins; Xenograft Model Antitumor Assays

A ruthenium(II) complex [Ru(p-cymene)(NHC)Cl2] (NHC=1,3-bis(4-(tert-butyl)benzylimidazol-2-ylidene), referred to as L-4, has been designed and synthesized recently in order to look for new anticancer drugs with high efficacy and low side effects. The anticancer activity and mechanism of action of L-4 in human esophageal squamous carcinoma EC109 cells were systematically investigated. The results revealed that L-4 exerted strong inhibitory effect on the proliferation of EC109 cells, and it arrested EC109 cells at G2/M phase, accompanied with the up-regulation of p53 and p21 and the down-regulation of cyclin D1. The results also showed that the reactive oxygen species (ROS)-dependent apoptosis of EC109 can be induced by L-4 via inhibiting the activity of glutathione reductase (GR), decreasing the ratio of glutathione to oxidized glutathione (GSH/GSSG), and leading to the generation of reactive oxygen species. The mitochondria-mediated apoptosis of EC109 induced by L-4 was also observed from the increase of Bax/Bcl-2 ratio, overload of Ca(2+), disruption of mitochondrial membrane potential (MMP), redistribution of cytochrome c, and activation of caspase-3/-9. However, the effects of L-4 on the cell viability, GR activity, GSH/GSSG ratio, reactive oxygen species level, mitochondria dysfunction and apoptosis induction were remarkably attenuated by adding the reactive oxygen species scavenger, NAC. Therefore, it was concluded that L-4 can inhibit the proliferation of EC109 cells via blocking cell cycle progression and inducing reactive oxygen species-dependent and mitochondria-mediated apoptosis. These findings suggested that the ruthenium(II) complex might be a potential effective chemotherapeutic agent for human esophageal squamous carcinoma (ESCC) and worthy of further investigation.

Topics: Antineoplastic Agents; Apoptosis; bcl-2-Associated X Protein; Calcium; Carcinoma, Squamous Cell; Caspase 3; Caspase 9; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cytochromes c; Down-Regulation; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Humans; Intracellular Space; Membrane Potential, Mitochondrial; Models, Molecular; Molecular Conformation; Organometallic Compounds; Protein Transport; Proto-Oncogene Proteins c-bcl-2; Ruthenium