FK228 potentiates topotecan activity against small cell lung cancer cells via induction of SLFN11.

The response rate of topotecan, as a second-line chemotherapeutic drug for small cell lung cancer, is ~20%. DNA/RNA helicase SLFN11 (schlafen family member 11), a member of the Schlafen (SLFN) family, is a crucial determinant of response to many DNA damaging agents, expression of SLFN11 tends to augment the antitumor effects of the commonly used DNA-targeting agents. In the present study we investigated how SLFN11 expression regulated the sensitivity of small cell lung cancer to topotecan. We showed that SLFN11 expression levels were positively associated with the sensitivity to topotecan in a panel of seven SCLC cell lines. Topotecan treatment induced different patterns of the DNA response network in SCLC cells: DNA damage response (DDR) was more prominently activated in SLFN11-deficient SCLC cell line H82 than in SLFN11-plentiful SCLC cell line DMS273, whereas topotecan induced significant accumulation of p-Chk1, p-RPA2 and Rad51 in H82 cells, but not in DMS273 cells. We unraveled that SLFN11 expression was highly negatively correlated to the methylation of the SLFN11 promoter. HDAC inhibitors FK228 and SAHA dose-dependently increased SLFN11 expression through suppressing DNA methylation at the SLFN11 promoter, thereby sensitizing SCLC cells to topotecan. Finally, we assessed the methylation status of the SLFN11 promoter in 27 SCLC clinical specimens, and found that most of the clinical samples (24/27) showed DNA methylation at the SLFN11 promoter. In conclusion, it is feasible to combine topotecan with FK228 to improve the response rate of topotecan in SCLC patients.

Combinations of proteasome inhibitors with obatoclax are effective for small cell lung cancer.

Proteasome inhibitors, bortezomib (BTZ), and carfilzomib (CFZ) are approved drugs for hematological malignancies, but lack anticancer activities against most solid tumors. Small cell lung cancer (SCLC) is a very aggressive neuroendocrine carcinoma of the lungs demanding effective therapy. In this study we investigated whether BTZ or CFZ combined with obatoclax (OBX), an antagonist for MCL-1 and a pan-BCL family inhibitor, could cause synergistic growth inhibition of SCLC cells. We showed that combined application of BTZ or CFZ with OBX caused synergistic growth inhibition of human SCLC cell lines (H82, H526, DMS79, H196, H1963, and H69) than single agent alone. Both BTZ-OBX and CFZ-OBX combinations displayed marked synergism on inducing apoptosis (~50% increase vs BTZ or CFZ alone). A comprehensive proteomics analysis revealed that BTZ preferentially induced the expression of MCL-1, an antiapoptotic protein, in SCLC cells. Thus, proteasome inhibitor-OBX combinations could specifically induce massive growth inhibition and apoptosis in SCLC cells. Subsequent proteome-wide profiling analysis of activated transcription factors suggested that BTZ- or CFZ-induced MCL-1 upregulation was transcriptionally driven by FOXM1. In nude mice bearing in SCLC H82 xenografts, both BTZ-OBX, and CFZ-OBX combinations exhibited remarkable antitumor activities against SCLC tumors evidenced by significant reduction of tumor size and the proliferation marker Ki-67 signals in tumor tissues as compared with single agent alone. Thus, proteasome inhibitor-OBX combinations are worth immediate assessments for SCLC in clinical settings.

Chlorinated effluent organic matter causes higher toxicity than chlorinated natural organic matter by inducing more intracellular reactive oxygen species.

During unplanned indirect potable reuse, treated wastewater that contains effluent organic matter (EOM) enters the drinking water source, resulting in different toxicity from natural organic matter (NOM) in surface water during chlorination. This study found that, during chlorination, EOM formed more total organic halogen (TOX) and highly toxic nitrogenous disinfection byproducts (DBPs) like dichloroacetonitrile and trichloronitromethane than NOM did. Oxidative stress including both reactive oxygen species (ROS) and reactive nitrogen species (RNS) in Chinese hamster ovary (CHO) cells substantially increased when exposed to chlorinated EOM and chlorinated NOM. The excessive ROS damaged biological macromolecules including DNA, RNA to form 8-hydroxy-(deoxy)guanosine and proteins to form protein carbonyls. Impaired macromolecule further triggered cell cycle arrest at the S and G2 phases, led to cell apoptosis and eventual necrosis. Cytotoxicity and genotoxicity of chlorinated EOM were both higher than those of chlorinated NOM. Adding the blocker L-buthionine-sulfoximine of intracellular antioxidant glutathione demonstrating that oxidative stress might be responsible for toxicity. ROS was further identified to be the main cause of toxicity induction. These findings highlight the risk from chlorinated EOM in the case of unplanned indirect potable reuse, because it showed higher level of cytotoxicity and genotoxicity than chlorinated NOM via inducing more ROS in mammalian cells.

Electron donating capacity reduction of dissolved organic matter by solar irradiation reduces the cytotoxicity formation potential during wastewater chlorination.

After treated wastewater is discharged into surface water for unplanned indirect potable reuse, solar irradiation transforms the dissolved organic matter (DOM), which would alter the formation of disinfection byproducts (DBPs) and change the cytotoxicity formation potential (CtFP) during post-chlorination in drinking water treatment plants. This study investigated the effects of solar irradiation on the CtFP and total organic halogen formation potential (TOXFP) of wastewater during post-chlorination. Exposure to natural sunlight decreased the formation potential of cytotoxicity to Chinese Hamster Ovary cells. Under 24 h simulated solar irradiation, CtFP and TOXFP decreased by more than 40%. X-ray photoelectron spectra and Fourier transformation infrared spectra suggested solar irradiation destroyed the key DBP precursors containing phenolic hydroxyl moieties (Ph-OH). The destruction of Ph-OH under solar irradiation was reflected by a decrease in the electron donating capacity (EDC) of DOM and the post-chlorination decreased the EDC further. Increasing the irradiation-consumed EDC abated the chlorine-consumed EDC, while the chlorine-consumed EDC was positively correlated to the CtFP and TOXFP by means of the electrophilic substitution-aromatic ring cleavage. Solar irradiation thus reduced the CtFP and TOXFP in wastewater during post-chlorination. This study revealed that solar irradiation decreased the risks of treated wastewater for unplanned indirect potable reuse and provided a strategy of controlling CtFP and TOXFP via reducing EDC of DOM in pretreatments.

Exposure to solar light reduces cytotoxicity of sewage effluents to mammalian cells: Roles of reactive oxygen and nitrogen species.

Sewage effluents can contain hundreds of toxic pollutants, making them a risk to humans when involved in drinking water. It is therefore important to evaluate the cytotoxicity of sewage effluents to mammalian cells. Solar light might influence the water quality of sewage effluents after their discharge into lakes or rivers, altering their cytotoxicity. In this study, natural solar light was found to lower the cytotoxicity of sewage effluents to Chinese hamster ovary (CHO) cells. Cytotoxicity of different samples decreased by 31%-65% after 12 h of simulated irradiation. Ultraviolet in sunlight was the major contributor to the cytotoxicity reduction. Aquatic reactive oxygen species (ROS), including singlet oxygen, superoxide anions, hydrogen peroxide, and hydroxyl radicals, were generated in the effluents under irradiation and they contributed to part of cytotoxicity reduction. Pollutants in sewage effluents induced cytotoxicity by simultaneously elevating the levels of intracellular ROS and intracellular reactive nitrogen species (RNS) in CHO cells. Solar light and the aquatic ROS formed under irradiation reduced the cytotoxicity because the transformed pollutants in sewage effluents increased lower intracellular ROS and RNS levels. These results help reveal the detoxification mechanism of sewage effluents in natural environment.

Formation and control of disinfection byproducts and toxicity during reclaimed water chlorination: A review.

Chlorination is essential to the safety of reclaimed water; however, this process leads to concern regarding the formation of disinfection byproducts (DBPs) and toxicity. This study reviewed the formation and control strategies for DBPs and toxicity in reclaimed water during chlorination. Both regulated and emerging DBPs have been frequently detected in reclaimed water during chlorination at a higher level than those in drinking water, indicating they pose a greater risk to humans. Luminescent bacteria and Daphnia magna acute toxicity, anti-estrogenic activity and cytotoxicity generally increased after chlorination because of the formation of DBPs. Genotoxicity by umu-test and estrogenic activity were decreased after chlorination because of destruction of toxic chemicals. During chlorination, water quality significantly impacted changes in toxicity. Ammonium tended to attenuate toxicity changes by reacting with chlorine to form chloramine, while bromide tended to aggravate toxicity changes by forming hypobromous acid. During pretreatment by ozonation and coagulation, disinfection byproduct formation potential (DBPFP) and toxicity formation potential (TFP) occasionally increase, which is accompanied by DOC removal; thus, the decrease of DOC was limited to indicate the decrease of DBPFP and TFP. It is more important to eliminate the key fraction of precursors such as hydrophobic acid and hydrophilic neutrals. During chlorination, toxicities can increase with the increasing chlorine dose and contact time. To control the excessive toxicity formation, a relatively low chlorine dose and short contact time were required. Quenching chlorine residual with reductive reagents also effectively abated the formation of toxic compounds.

Solar light irradiation significantly reduced cytotoxicity and disinfection byproducts in chlorinated reclaimed water.

Chlorinated reclaimed water is widely used for landscaping and recreational purposes, resulting in human exposure to toxic disinfection byproducts. Although the quality of chlorinated reclaimed water might be affected by sunlight during storage, the effects of solar light irradiation on the toxicity remain unknown. This study investigated the changes in cytotoxicity and total organic halogen (TOX) of chlorinated reclaimed water exposed to solar light. Irradiation with solar light for 12 h was found to significantly reduce the cytotoxicity of chlorinated reclaimed water by about 75%, with ultraviolet light being responsible for the majority of this reduction. Chlorine residual in reclaimed water tended to increase the cytotoxicity, and the synergy between solar light and free chlorine could not enhance the reduction of cytotoxicity. Adding hydroxyl radical scavengers revealed that the contribution of hydroxyl radical to cytotoxicity reduction was limited. Solar light irradiation concurrently reduced TOX. The low molecular weight (<1 kDa) fraction was the major contributor of cytotoxicity and TOX in chlorinated reclaimed water. Detoxification of the low molecular weight fraction by light irradiation was mainly a result of TOX dehalogenation, while detoxification of the high molecular weight (>1 kDa) fraction was probably caused by photoconversion from high toxic TOX to low toxic TOX.