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5-HT3 receptor antagonists induce autophagy via extracellular signal-related kinase (ERK) activation in lung cancer cells. (A) Levels of phospho-ERK, total-ERK, light chain 3B (LC3B), and autophagy-related 16 like 1 (ATG16L1) were determined by Western blotting. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as a loading control. (B) A549 cells were treated with or without ERK inhibitor (U0126, 10 mM) before a 1-h treatment with 5-HT3 receptor antagonist. Levels of phospho-ERK, total-ERK, and LC3B were determined by Western blotting. GAPDH served as a loading control; * p < 0.05.
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Unlike 5-hydroxytryptamine (5-HT, serotonin) 1 and 5-HT2, the effect of 5-HT3 receptors on tumor cells is poorly understood. We conducted this study to determine whether the perioperative use of 5-HT3 receptor antagonists, which are widely used antiemetics, impacts the recurrence and mortality after lung cancer surgery and related anti-tumor mechan...
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... In a retrospective study, Park et. al. reported that the use of palonosetron or ramosetron were both associated with a significantly lower rate of reoccurrence lung cancer surgery (Lee, Park, and Kim, 2019). Their data indicated both palonosetron and ramosetron acted as novel autophagy inducers, suggested that the enhancement of this pathway may accelerate degradation of damaged cells. ...
Space travel increases galactic cosmic ray exposure to flight crews and this is significantly elevated once travel moves beyond low Earth orbit. This includes combinations of high energy protons and heavy ions such as ⁵⁶Fe or ¹⁶O. There are distinct differences in the biological response to low-energy transfer (x-rays) or high-energy transfer (High-LET). However, given the relatively low fluence rate of exposure during flight operations, it might be possible to manage these deleterious effects using small molecules currently available. Virtually all reports to date examining small molecule management of radiation exposure are based on low-LET challenges. To that end an FDA approved drug library (725 drugs) was used to perform a high throughput screen of cultured cells following exposure to galactic cosmic radiation. The H9c2 myoblasts, ES-D3 pluripotent cells, and Hy926 endothelial cell lines were exposed to a single exposure (75 cGy) using the 5-ion GCRsim protocol developed at the NASA Space Radiation Laboratory (NSRL). Following GCR exposure cells were maintained for up to two weeks. For each drug (@10µM), a hierarchical cumulative score was developed incorporating measures of mitochondrial and cellular function, oxidant stress and cell senescence. The top 160 scores were retested following a similar protocol using 1µM of each drug. Within the 160 drugs, 33 are considered to have an anti-inflammatory capacity, while others also indirectly suppressed pro-inflammatory pathways or had noted antioxidant capacity. Lead candidates came from different drug classes that included angiotensin converting enzyme inhibitors or AT1 antagonists, COX2 inhibitors, as well as drugs mediated by histamine receptors. Surprisingly, different classes of anti-diabetic medications were observed to be useful including sulfonylureas and metformin. Using a hierarchical decision structure, we have identified several lead candidates. That no one drug or even drug class was completely successful across all parameters tested suggests the complexity of managing the consequences of galactic cosmic radiation exposure.
... In a retrospective study, Park et. al. reported that the use of palonosetron or ramosetron were both associated with a significantly lower rate of reoccurrence lung cancer surgery (Lee, Park, and Kim, 2019). Their data indicated both palonosetron and ramosetron acted as novel autophagy inducers, suggested that the enhancement of this pathway may accelerate degradation of damaged cells. ...
Space exploration involves many dangers, including radiation exposure from galactic cosmic radiation (GCR). This includes high energy protons and heavy ionizing ions such as 56 Fe, 28 Si, and 16 O. NASA has defined GCR as a carcinogenic risk for long-duration space missions. To date no clear strategy has been developed to counter chronic GCR exposure. We hypothesize that preconditioning cells with low levels of radiation will be protective from subsequent higher radiation exposures. H9C2 cells maintained in culture were pretreated with 0, 0.1, or 0.5 Gy X-rays using a RadSource 2000 Irradiator. The challenge radiation treatment consisted of either 8 Gy X-ray or 75cGy of GCR, using a 5 Ion GCRsim protocol. To determine cell health, the cell doubling time assay was used. 10000 cells from each treatment group were put into wells of a 24 well plate and maintained for 48-72 hours. Following harvest, cells were counted using a flow cytometer. The 8 Gy X-ray challenge alone significantly (p<.05) increased cell doubling time compared to the no radiation control group. The radiation pre-treatment groups ameliorated the 8 Gy X-ray induced increases in cell doubling time and there was no difference compared to the no radiation control group. 75 cGy GCR challenge alone significantly increased cell doubling time compared to the no radiation group. Following the 75cGy challenge, only the 0.5 Gy pre-treatment ameliorated the 75cGy induced increases in cell doubling time. The cell doubling time protocol measures the amount of time require for a cell to undergo mitosis. It is directly influence by DNA integrity as well as mitochondrial and cellular function. DNA damage or pathologic oxidant stress will delay the replicative functions. As such this is a physiologic measure of the cell wellbeing. Our results suggested that pretreatment with low dose X-rays enhanced cellular resiliency by inducing an adaptive response which offered a small but significant protection against a following higher radiation challenge. Although perhaps not a practical countermeasure, these findings may serve to offer insight to cell signaling pathways activated in response to low dose irradiation and that might be targeted as a potential countermeasure.
... In a retrospective study, Park et. al. reported that the use of palonosetron or ramosetron were both associated with a significantly lower rate of reoccurrence lung cancer surgery (Lee, Park, and Kim, 2019). Their data indicated both palonosetron and ramosetron acted as novel autophagy inducers, suggested that the enhancement of this pathway may accelerate degradation of damaged cells. ...
Galactic cosmic radiation (GCR) exposure is increased to space flight crews as travel moves beyond low Earth orbit. GCR includes high energy protons and heavy ionizing ions such as ⁵⁶ Fe, ²⁸ Si, and ¹⁶ O and there are distinct differences in the biological response compared to low‐energy transfer events (x‐rays). Although the exposure rate is relatively low, the cumulative radiation dose is likely to have deleterious effects for long duration flight crews. The underlying mechanisms for these elevated risks are thought to include direct DNA damage as well as chronically elevated oxidant stress, and inflammation.
Given the relatively low fluence rate of GCR exposure during flight operations it might be possible to manage these deleterious effects using small molecules currently available. To that end we have utilized a FDA approved drug library to perform a high throughput screen of cultured cells following exposure to GCR. H9c2 cells were exposed to a single 75 cGy dose using the 5‐ion GCRsim protocol developed by NSRL. Following GCRsim exposure, cells were treated using 10 µM for each drug. At harvest, cells were tested for protein content, oxidant stress, cellular senescence, and mitochondrial function.
Of 725 drugs, more than 500 were ineffective in managing GCR exposure. Within the top 160 drugs, 33 were primarily designated as anti‐inflammatory. Significant variations within drug classes were observed. Only 10 of 46 drugs that interacted with histamine receptors were found useful and most were 5‐HT3 specific. The COX inhibitors deemed useful were all COX2 specific. Surprisingly, some anti‐diabetic medications observed to be useful including metformin.
The bias of mitochondrial degradation as a central readout comes from several “omics” analyses that identified mitochondrial dysfunction as a consequence of GCR exposure. Mitochondrial dysfunction is also an antecedent event in the transformation leading to carcinogenesis. And the potential for carcinogenesis is a significant risk for long duration space missions. Separate from examining mitochondrial and cellular dysfunction as readouts, we have developed a Afp‐tdTomato expressing cell line that is responsive to different carcinogenic reagents including GCR. We have observed that some of the lead candidates will at least partially rescue the cells from GCR induced Afp driven expression. Additionally adapting cell doubling time protocols we observed that GCR exposure significantly increased cell doubling time; a finding indicative of mitotic dysfunction. We also observed that this increase was partially ameliorated by some of the lead candidates. That no one class of drug dominated the response to radiation exposure speaks to the complexity of managing GCR exposure.
... The 5-HT3RAs used such as ondansetron are highly selective and preclinical studies in rat have shown the effects of small doses of ondansetron on cognition, behavioural sensitisation, and epilepsy (Kwan et al., 2020). In a recent study, perioperative use of 5-HT3RAs (palonosetron or ramosetron) is associated with increased recurrence-free survival in patients after thoracotomy for lung cancer although the doses used were not presented in detail and ondansetron was excluded from the 5-HT3RAs (Lee et al., 2019). It is suggested that small doses of 5-HT3RAs may also exhibit peripheral effect. ...
Liver failure is a severe clinical syndrome with high mortality. 5-Hydroxytryptamine 3 receptor antagonists (5-HT3RAs) can reduce liver damage in animal models. We investigated whether 5-HT3RAs may improve the prognosis of liver failure. We analyzed the 28 and 90 days mortality of liver failure patients in relation to the use of 5-HT3RAs using data from a tertiary hospital in northwest China. According to the use of 5-HT3RAs, 419 patients with liver failure (46 acute, 93 sub-acute, 44 chronic, 236 acute on chronic) were divided into 5-HT3RA group (n = 105) and control group (n = 314). 5-HT3RAs were associated with decreased 28 days (HR 0.18, 95% CI 0.10-0.34, p < 0.001) and 90 days (HR 0.21, 95% CI 0.13-0.33, p < 0.001) mortality. After propensity score matching (PSM) (n = 67 in each group), 5-HT3RAs were still significantly associated with reduced 28 days (HR 0.10, 95%CI 0.04-0.26, p < 0.001) and 90 days (HR 0.16, 95%CI 0.08-0.31, p < 0.001) mortality. 5-HT3RA group patients had significantly higher 28 and 90 days survivals than controls both before and after PSM (all p < 0.001). This study shows that 5-HT3RAs are associated with increased survival of liver failure patients and thus may be used to treat liver failure if the findings are confirmed by additional studies.
... 5 Although 5-HT3 receptor is expressed mainly throughout the central and peripheral nervous systems, some papers report the effect of 5-HT3 receptors on tumor cells. Lee et al 6 In this study, we found that HTR3A was preferentially expressed in lung adenocarcinoma with aggressive histological subtypes by RNA sequencing and revealed that HTR3A expression was related to histologic subtypes in lung adenocarcinoma. By immunohistochemical analysis of 63 lung adenocarcinoma cases, we showed the higher expression level of HTR3A was detected in acinar, papillary, and solid adenocarcinoma than in AIS and lepidic adenocarcinoma; the former was a more aggressive subtype than the latter. ...
... receptor is related to cancer progression in some types of cancer including lung carcinoma. 6,7 To date, no studies have compared HTR3A expression in histologic subtypes of lung adenocarcinoma. In the present study, we identified HTR3A, which was predominantly expressed in the poor-prognosis group, and found that invasive size Lung adenocarcinoma has various histologic subtypes. ...
Lung cancer is the leading cause of cancer death around the world. Adenocarcinoma is the most common histological type and has various histologic subtypes: lepidic, acinar, papillary, solid, and invasive mucinous adenocarcinoma. Histologic subtypes are related to invasiveness of tumors; for example, lepidic subtype is less invasive than acinar/papillary subtype. HTR3A is the main subunit of 5‐hydroxytryptamine 3 (5‐HT3) receptors, which are the only ligand‐gated ion channels in 7 families of 5‐HT receptors. Though 5‐HT3 receptor is expressed mainly throughout the central and peripheral nervous systems, some papers report the effect of 5‐HT3 receptors on tumor cells including lung cancer. However, whether HTR3A correlates with histopathological findings such as the histologic subtypes or the distribution in an individual sample remains unclear. Immunohistochemically, we revealed that the higher expression level of HTR3A was detected in acinar, papillary, and solid adenocarcinoma than in adenocarcinoma in situ and lepidic adenocarcinoma; the former was more aggressive subtype than the latter. We also showed the relationship of HTR3A expression to Ki‐67 positivity, widely used as a proliferation marker. Moreover, we generated HTR3A‐knockdown lung adenocarcinoma cells and showed that the HTR3A knockdown attenuated proliferation via ERK phosphorylation. Our results demonstrated that HTR3A expression was related to proliferation in lung adenocarcinoma by means of both in vitro and immunohistochemical assays on clinical samples. We showed the therapeutic potential of a 5‐HT3 receptor antagonist, tropisetron, for the treatment of lung adenocarcinoma.
The development of a scalable and highly reproducible in vitro tumor microenvironment (TME) platform still sheds light on new insights into cancer metastasis mechanisms and anticancer therapeutic strategies. Here, we present an all‐in‐one injection molded plastic array 3D culture platform (All‐in‐One‐IMPACT) that integrates vascularized tumor spheroids for highly reproducible, high‐throughput experimentation. This device allows the formation of self‐assembled cell spheroids on a chip by applying the hanging drop method to the cell culture channel. Then, when the hydrogel containing endothelial cells and fibroblasts is injected, the spheroid inside the droplet can be patterned together in three dimensions along the culture channel. In just two steps above, we can build a vascularized TME within a defined area. This process does not require specialized user skill and minimizes error‐inducing steps, enabling both reproducibility and high‐throughput of the experiment. We have successfully demonstrated the process, from spheroid formation to tumor vascularization, using patient‐derived cancer cells (PDCs) as well as various cancer cell lines. Furthermore, we performed combination therapies with Taxol (paclitaxel) and Avastin (bevacizumab), which are used in standard care for metastatic cancer. The All‐in‐One IMPACT is a powerful tool for establishing various anticancer treatment strategies through the development of a complex TME for use in high‐throughput experiments. This article is protected by copyright. All rights reserved.
5-hydroxytryptamine type 3 (5-HT3) receptors are ligand gated ion channels, which clearly distinguish their mode of action from the other G-protein coupled 5-HT or serotonin receptors. 5-HT3 receptors are well established targets for emesis and gastrointestinal mobility and are used as adjunct targets in treating schizophrenia. However, the distribution of these receptors is wider than the nervous system and there is potential that these additional sites can be targeted to modulate inflammatory and/or metabolic conditions. Recent progress in structural biology and pharmacology of 5-HT3 receptors have provided profound insights into mechanisms of their action. These advances, combined with insights into clinical relevance of mutations in genes encoding 5-HT3 subunits and increasing understanding of their implications in patient’s predisposition to diseases and response to the treatment, open new avenues for personalized precision medicine. In this review, we recap on the current status of 5-HT3 receptor-based therapies using a biochemical and physiological perspective. We assess the potential for targeting 5-HT3 receptors in conditions involving metabolic or inflammatory disorders based on recent findings, underscoring the challenges and limitations of this approach.
Serotonin (5-hydroxytryptamine, 5-HT) metabolism has long been linked to tumorigenesis and tumor progression. Numerous studies have shown the functions of 5-HT and its metabolites in the regulation of tumor biological processes like cell proliferation, invasion, metastasis, tumor angiogenesis and immunomodulatory through multi-step complex mechanisms. Reprogramming of 5-HT metabolism has been revealed in various tumors paving way for development of drugs that target enzymes, metabolites or receptors involved in 5-HT metabolic pathway. However, information on the role of 5-HT metabolism in cancer is scanty. This review briefly describes the main metabolic routes of 5-HT, the role of 5-HT metabolism in cancer and systematically summarizes the most recent advances in 5-HT metabolism-targeted cancer therapy.
