10<em>H</em>-3,6-Diazaphenothiazine induces G₂/M phase cell cycle arrest and caspase-dependent apoptosis and inhibits cell invasion of A2780 ovarian carcinoma cells through the regulation of NF-&kappa;B and (BIRC6-XIAP) complexes

Abstract

The asymptomatic properties and high treatment resistance of ovarian cancer result in poor treatment outcomes and high mortality rates. Although the fundamental chemotherapy provides promising anticancer activities, it is associated with severe side effects. The derivative of phenothiazine, namely, 10H-3,6-diazaphenothiazine (PTZ), was synthesized and reported with ideal anticancer effects in a previous paper. In this study, detailed anticancer properties of PTZ was examined on A2780 ovarian cancer cells by investigating the cytotoxicity profiles, mechanism of apoptosis, and cell invasion. Research outcomes revealed PTZ-induced dose-dependent inhibition on A2780 cancer cells (IC50 =0.62 µM), with significant less cytotoxicity toward HEK293 normal kidney cells and H9C2 normal heart cells. Generation of reactive oxygen species (ROS) and polarization of mitochondrial membrane potential (ΔΨm) suggests PTZ-induced cell death through oxidative damage. The RT2 Profiler PCR Array on apoptosis pathway demonstrated PTZ-induced apoptosis via intrinsic (mitochondria-dependent) and extrinsic (cell death receptor-dependent) pathway. Inhibition of NF-κB and subsequent inhibition of (BIRC6-XIAP) complex activities reduced the invasion rate of A2780 cancer cells penetrating through the Matrigel™ Invasion Chamber. Lastly, the cell cycle analysis hypothesizes that the compound is cytostatic and significantly arrests cell proliferation at G2/M phase. Hence, the exploration of the underlying anticancer mechanism of PTZ suggested its usage as promising chemotherapeutic agent.

Combination of Ruthenium Complex and Doxorubicin Synergistically Inhibits Cancer Cell Growth by Down-Regulating PI3K/AKT Signaling Pathway

“…X-chromosome-linked IAP (XIAP), one of the eight human IAP proteins, has been found to exert the most pronounced antiapoptotic function, which is associated with its ability to block caspase−3, −7, and −9 (61). Moreover, some studies have demonstrated that XIAP has also been implicated in the regulation of NF-κB (62). In a previous study, it was found that -Ru1 induces apoptosis through the mitochondrial-mediated pathway, and regulates the expression of proapoptotic proteins, including the downregulation of Bcl-2/Bcl-xl, and the up-regulation of cytochrome c (Cyt C), caspase 9 and caspase 3 (32).…”

10H‐3,6‐Diazaphenothiazines triggered the mitochondrial‐dependent and cell death receptor‐dependent apoptosis pathways and further increased the chemosensitivity of MCF‐7 breast cancer cells via inhibition of AKT1 pathways

“…The tested compound for present study, namely PTZ was the tested compound preceded from previous studies. [19] Phenothiazine derivative was synthesized by professional medicinal chemistry research group from Medical University of Silesia, Sosnowiec, Republic of Poland; and this compound was synthesized according to the information obtained from respective publications. The detailed characteristic of this compound was explained in respective reports.…”

“…[15][16][17] Beyond the routine metabolic functions of estrogen hormone, binding of the estrogen to its membrane-bound receptors (ESR1 and ESR2) tends to produce several downstream targeted genes, which are generally involved in regulation of cell division and proliferation, inhibit cell death mechanism, and promote secondary metastasis of cancer cells. [18][19][20] It is commonly known that dietary factors such as high glucose diet intake would increase the risk to have breast cancer. [21] In cellular biology, the AKT1 gene detected presence of high glucose molecules and send the signals to pancreas for secretion of insulin hormone for glycolysis process; despite high amount of insulin eventually promoting cells to synthesize more insulin-like growth factor-1 receptor (IGF-1R) on surface of plasma membrane for cellular intake of glucose and subsequent glycolysis process.…”

“…Unfortunately, estrogen hormone binds nonselectively toward IGF-1R instead of its own estrogen receptor-1 (ESR1) or -2 (ESR2) and results in activation of cell proliferation signalling cascade and promoting cell survival via inhibition of programmed cell death mechanism. [19] In contrast, high mitochondrial thioredoxin reductase (TrxR) enzymatic activity boosts the mitochondrial activities of cancer cells and contributes to reduction in efficacy of chemotherapeutic compounds, thus yielding to resistance features of breast cancer. [20] Excessive electron sources surrounding the cancer cells eventually provided enough electron supplies for cancer cells' mitochondria to carry out routine tasks: (a) reduction of thioredoxin, the substrate of TrxR enzyme; (b) electron transport chain reaction for ATP production, which encouraged proliferation of cancer cells.…”

“…[24] Previous studies on anticancer activities of phenothiazine derivative compound, namely 10H-3,6-diazaphenothiazine (named as PTZ, for this report) on A2780 ovarian cancer cells exhibited inhibition on proliferation of cancer cells by induction of G 2 /M-phase cell cycle checkpoint and induced apoptosis via inhibitory regulation on NF-κB gene and BIRC6-XIAP complexes. [19] For instance, ovarian and breast cancers were both cancers related estrogen hormone and the interaction of estrogen hormone with several pro-proliferation membrane-bound receptor; this steroid-derived hormone enhances conjugation of NF-κB gene with BIRC gene family, for example, NF-κB-XIAP-BIRC6 complex. Thus leading to the inhibition of caspase activation, negative regulation of apoptosis pathways and contribute to the death signals resistance.…”