Risk factors for enhanced recovery after surgery failure in patients undergoing lung cancer resection with concomitant cardiovascular disease: A single-center retrospective study.

Objective: Enhanced recovery after surgery (ERAS) has been widely used in patients with lung cancer, and its effectiveness has been confirmed; however, some lung cancers with poor clinical outcomes lead to ERAS failure after radical resection. This study aimed to analyze risk factors associated with ERAS failure after radical resection in patients with lung cancer and concomitant cardiovascular disease. Methods: In total, 198 patients who underwent ERAS following radical lung cancer surgery for concomitant cardiovascular disease between January 2022 and September 2023 were enrolled in this retrospective study. The patients were categorized into two groups based on the definition of ERAS failure: ERAS success group (n = 152) and ERAS failure group (n = 46). Univariate and multivariate analyses were performed to investigate the risk factors of ERAS failure. Results: Univariate analysis showed that gender, tumor location, operation time, estimated blood loss (EBL), suction drainage, and total cholesterol were associated with ERAS failure. Multivariate analysis showed that operation time (odds ratio [OR] = 1.015; P = 0.011) and suction drainage (OR = 3.343; P = 0.008) were independent risk factors for ERAS failure. Conclusions: Operation time and suction drainage were independent risk factors for ERAS failure after radical resection of combined cardiovascular lung cancer. Therefore, improving surgical efficiency and postoperative chest drain management are important for successful ERAS.

Summary of best evidence for enhanced recovery after surgery for patients undergoing lung cancer operations.

According to the cancer burden report released by the International Agency for Research on Cancer (IARC) in 2020, the mortality rate of lung cancer is 18%, ranking first in the world, and its morbidity and mortality rates are highest in China. Pneumonectomy is the preferred treatment for lung cancer patients, but surgery carries a significant risk of perioperative complications, which may affect the patient's functional recovery and quality of life. So, the rehabilitation of the large number of lung cancer patients in China requires greater attention. A number of studies have shown that the enhanced recovery after surgery (ERAS) protocol can reduce the risk of death, readmission rate, adjuvant chemotherapy time, postoperative pain level, anesthesia medication amount, length of stay, and hospitalization expenses. Foreign literature has successively issued guidelines to improve recovery among lung cancer patients, but Chinese-specific literature for patients undergoing lung cancer surgery or thoracic surgery remains inadequate. Some Chinese expert consensus have only considered part of the content of ERAS in thoracic surgery. To summary the evidence of the ERAS program for lung cancer surgery patients at home and abroad basing on evidence-based medicine is necessary. Therefore, this study used evidence-based practical thinking as a guide to (1) evaluate, integrate, and summarize relevant evidence guidelines and data resources at home and abroad so as to construct an enhanced recovery program for lung cancer patients suitable for Chinese national conditions and (2) provide a scientific basis for future research and practice in related fields.

High VHL Expression Reverses Warburg Phenotype and Enhances Immunogenicity in Kidney Tumor Cells.

Clear cell renal cell carcinoma (ccRCC) is a frequently occurring renal cancer. The Von Hippel-Lindau disease tumor suppressor VHL, a known tumor suppressor gene, is frequently mutated in about 50% of patients with ccRCC. However, it is unclear whether VHL influences the progression of ccRCC tumors expressing wild-type VHL. In the present study, we found that higher expression of VHL was correlated with the better disease-free survival (DFS) in ccRCC patients using The Cancer Genome Atlas (TCGA) datasets. We revealed that VHL overexpression in ccRCC cells inhibited epithelial-mesenchymal transition (EMT), sterol regulatory element-binding protein 1 (SREBP1)-regulated triglyceride synthesis, and cell proliferation. Proteomic analysis provided us a global view that VHL regulated four biological processes, including metabolism, immune regulation, apoptosis, and cell movement. Importantly, we found that VHL overexpression led to up-regulated expression of proteins associated with antigen processing and interferon-responsive proteins, thus rendering ccRCC cells more sensitive to interferon treatment. We defined an interferon-responsive signature (IRS) composed of ten interferon-responsive proteins, whose mRNA expression levels were positively correlated with DFS in ccRCC patients. Taken together, our results propose that the subset of ccRCC patients with high VHL expression benefit from immunotherapy.

HSP60-knockdown suppresses proliferation in colorectal cancer cells via activating the adenine/AMPK/mTOR signaling pathway.

Colorectal cancer (CRC) is the fourth most lethal cancer in the world. Heat shock protein 60 (HSP60), a mitochondrial chaperone that maintains mitochondrial proteostasis, is highly expressed in tumors compared with in paracancerous tissues, suggesting that high HSP60 expression benefits tumor growth. To determine the effects of HSP60 expression on tumor progression, stable HSP60-knockdown HCT116 cells were constructed in the present study, revealing that knockdown of HSP60 inhibited cell proliferation. Proteomic analysis demonstrated that mitochondrial proteins were downregulated, indicating that knockdown of HSP60 disrupted mitochondrial homeostasis. Metabolomic analysis demonstrated that cellular adenine levels were >30-fold higher in HSP60-knockdown cells than in control cells. It was further confirmed that elevated adenine activated the AMPK signaling pathway, which inhibited mTOR-regulated protein synthesis to slow down cell proliferation. Overall, the current results provide a valuable resource for understanding mitochondrial function in CRC, suggesting that HSP60 may be a potential target for CRC intervention.

HSP60-regulated Mitochondrial Proteostasis and Protein Translation Promote Tumor Growth of Ovarian Cancer.

Ovarian cancer (OC) is the most lethal gynecological carcinoma due to the lack of diagnostic markers and effective drug targets. Discovery of new therapeutic targets in OC to improve the treatment outcome is urgently needed. We performed proteomic analysis of OC specimens and the paired normal tissues and revealed that proteins associated with mitochondrial proteostasis and protein translation were highly expressed in ovarian tumor tissues, indicating that mitochondria are required for tumor progression of OC. Heat shock protein 60 (HSP60), an important mitochondrial chaperone, was upregulated in ovarian tumors. HSP60 silencing significantly attenuated growth of OC cells in both cells and mice xenografts. Proteomic analysis revealed that HSP60 silencing downregulated proteins involved in mitochondrial functions and protein synthesis. Metabolomic analysis revealed that HSP60 silencing resulted in a more than 100-fold increase in cellular adenine levels, leading to increased adenosine monophosphate and an activated AMPK pathway, and consequently reduced mTORC1-mediated S6K and 4EBP1 phosphorylation to inhibit protein synthesis that suppressed the proliferation of OC cells. These results suggest that HSP60 knockdown breaks mitochondrial proteostasis, and inactivates the mTOR pathway to inhibit OC progression, suggesting that HSP60 is a potential therapeutic target for OC treatment.

HSP60 silencing promotes Warburg-like phenotypes and switches the mitochondrial function from ATP production to biosynthesis in ccRCC cells.

HSP60 is a major mitochondrial chaperone for maintaining mitochondrial proteostasis. Our previous studies showed that HSP60 was significantly downregulated in clear cell renal cell carcinoma (ccRCC), the most common type of kidney cancer characterized by the classic Warburg effect. Here, we analyzed datasets in The Cancer Genome Atlas and revealed that higher HSP60 expression correlated with better overall survival in ccRCC patients. We also stably knocked down or overexpressed HSP60 in ccRCC cells to investigate the effects of HSP60 expression on the transition between oxidative phosphorylation and glycolysis. We confirmed that HSP60 knockdown increased cell proliferation, whereas its overexpression decreased cell growth. Proteomics and metabolomics revealed that HSP60 knockdown promoted Warburg-like phenotypes with enhanced glycolysis and decreased mitochondrial activity. Consistent with this finding, isotope tracing showed that the metabolic flow from glycolysis to TCA was reduced. However, HSP60 silencing enhanced mitochondrial functions in glutamine-directed biosynthesis with increased flow in two parts of the TCA cycle: Gln→αKG→OAA→Asp and Gln→αKG→ISO→acetyl-CoA, resulting in elevated de novo nucleotide synthesis and lipid synthesis. Proteomic analysis indicated that HSP60 silencing activated NRF2-mediated oxidative stress responses, while glutamate generated from glutamine increased glutathione synthesis for quenching excessive reactive oxygen species (ROS) produced upon elevated cell growth. We further found that HSP60 silencing activated the MEK/ERK/c-Myc axis to promote glutamine addiction, and confirmed that ccRCC cells were susceptible to oxidative stress and glutaminase inhibition. Collectively, our data show that HSP60 knockdown drives metabolic reprogramming in ccRCC to promote tumor progression and enhances mitochondrial-dependent biosynthesis.

Efficacy of Pain Relief in Different Postherpetic Neuralgia Therapies: A Network Meta-Analysis.

BACKGROUND: Postherpetic neuralgia (PHN) is a nerve pain disease usually controlled by different therapies, i.e., topical therapies, antiepileptics, analgesics, antipsychotics, antidepressants, anti-dementia drugs, antivirals, amitriptyline, fluphenazine, and magnesium sulfate. It is believed that different therapies may lead to different levels of pain relief. OBJECTIVES: We proposed this study to compare the efficacy of PHN treatments. STUDY DESIGN: We conducted a systematic review of the current literature. All relevant studies were retrieved from online databases. The standardized mean difference (SMD) was used for pain relief measurement in different PHN therapies. SETTING: A conventional meta-analysis and a network meta-analysis (NMA) were carried out together with the surface under the cumulative ranking curve (SUCRA) for each therapy calculated regarding their efficacy. RESULTS: A pairwise meta-analysis suggested that 4 treatment classes, including topical therapies, antiepileptics, analgesics, and antidepressants, exhibited better pain relief results than placebo. Likewise, a NMA suggested that patients with 4 treatment classes exhibited significant improvements in pain scores compared to those with placebo. LIMITATIONS: There is a lack of direct head-to-head comparisons of some treatments, especially for antivirals, anti-dementia drugs, and magnesium sulfate. Secondly, the specific agents belonging to the same class of therapies might exhibit different effects (gabapentin and carisbamate) with different mechanisms (opioids and ketamine) on reducing pain, and some agents were hard to find in literatures and were not involved in our study, which may influence our results. CONCLUSIONS: Analgesics were preferable to other treatments with respect to pain relief for PHN, while antivirals appeared to be less effective than other therapies. KEY WORDS: Postherpetic neuralgia, topical agents, antiepileptics, analgesics, antipsychotics, antidepressants.

Inhibition of NAMPT decreases cell growth and enhances susceptibility to oxidative stress.

Nicotinamide adenine dinucleotide (NAD) is an essential molecule for living organisms and plays a vital role in aging and age-associated diseases. In eukaryotic cells, cellular NAD is mainly generated by the scavenge pathway in which nicotinamide phosphoribosyltransferase (NAMPT) catalyzes the formation of nicotinamide mononucleotide. Inhibition of NAMPT is a therapeutic strategy for cancer treatment. To explore the effects of NAMPT inhibition on cellular processes, cells were treated with 10 nM FK866, an NAMPT inhibitor, resulting in a decrease in the cellular NAD level, a lower growth rate, and enhanced susceptivity to oxidative stress as compared to the untreated cells. Quantitative proteomics revealed that 325 proteins were downregulated in the FK866-treated cells, and were involved in diverse cellular processes including nucleobase-containing compound metabolic process, protein metabolic process, antioxidant and DNA repair processes. Downregulation of 4 selected proteins was confirmed by western blotting and quantitative PCR. Downregulation of antioxidant proteins GRX1 and catalase, and DNA-repair proteins PCNA and PARP1 contributed to the enhanced susceptibility of FK866-treated cells to oxidative stress. FK866 treatment also caused mitochondrial dysfunction through downregulation of mitochondrial ribosomal proteins. Taken together, these results demonstrate that FK866 treatment efficiently decreases the cellular NAD level and induces autonomous changes in proteostasis, leading to cell growth inhibition and increased susceptibility to oxidative stress.

Proteomic Analysis of eIF5B Silencing-Modulated Proteostasis.

Protein translational machinery is an important component of the proteostasis network that maintains cellular proteostasis and regulates aging and other cellular processes. Ample evidence indicates that inhibition of translation initiation factor activities enhances stress resistance in model organisms. Eukaryotic translation initiation factor 5B (eIF5B) acts by joining the pre-40S subunit with the 60S ribosomal unit to form an 80S-like complex during protein translational initiation. Reduced eIF5B expression may disrupt proteostasis and trigger cellular processes associated with stress responses. In this study, the physiological effects of altered eIF5B expression were examined in 293T and HepG2 cells. Cells with eIF5B-knockdown (eIF5B-KN) grew more slowly than control cells, and had a lower level of intracellular reactive oxygen species (ROS), increased resistance to oxidative stress and enhanced autophagy. Proteomic analysis showed that eIF5B knockdown resulted in upregulation of 88 proteins and downregulation of 130 proteins compared with control cells. The differentially expressed proteins were associated with diverse cellular processes including amino acid metabolism, RNA processing and protein metabolism, and DNA synthesis. Autonomous downregulation of the mitogen-activated protein kinase (MAPK) signaling pathway was identified as confirmed by western blotting and qPCR. We proposed that deactivation of MAPK pathway modulated proteostasis and induced prolonged S-phase of the cell-cycle, contributing to the slow growth of eIF5B-KN cells. eIF5B silencing also inactivated the mTOR pathway, downregulated glutamine transporters, enhanced autophagy, and decreased 28S rRNA and 5.8S rRNA expression levels which were reversed by restoration of eIF5B expression. Taken together, these results suggest that eIF5B silencing provides a negative feedback to deactivate MAPK signaling, leading to reduced cell growth. These findings provide a useful resource to further biological exploration of the functions of protein synthesis in regulation of proteostasis and stress responses.

Proteomic analysis of gossypol induces necrosis in multiple myeloma cells.

Gossypol is a phenolic aldehyde extracted from plants and is known to be an antitumor agent to induce cancer cell apoptosis. In the present study, multiple myeloma cells were treated with gossypol, which resulted in an increase of cellular reactive oxygen species (ROS) and cell necrosis. Quantitative proteomic analysis was carried out to identify differentially expressed proteins between untreated and gossypol-treated cells. Proteomic analysis identified 4330 proteins, in which 202 proteins are upregulated and 383 proteins are downregulated in gossypol-treated cells as compared to the untreated cells. Importantly, proteomic and western blot analysis showed that apoptosis regulators BAK and Bax were upregulated in gossypol-treated cells, indicating that Bcl-2 associated death pathway was activated. Similarly, gossypol also induced upregulations of DNA mismatch repair proteins and DNA replication licensing factor, suggesting that gossypol caused significant DNA damage. Furthermore, upregulations of HLA class I and class II histocompatibility antigens and beta-2-microglobulin were observed in gossypol-treated cells, indicating that gossypol has a novel function to activate cellular immune responses. Our data demonstrate that the execution of necrosis is a complex process involving ROS, DNA damage, and Bcl-2 family proteins. Gossypol-activated immune responses are a potential new approach for multiple myeloma chemotherapy.

Azacytidine induces necrosis of multiple myeloma cells through oxidative stress.

Azacytidine is an inhibitor of DNA methyltransferase and is known to be an anti-leukemic agent to induce cancer cell apoptosis. In the present study, multiple myeloma cells were treated with azacytidine at clinically relevant concentrations to induce necrosis through oxidative stress. Necrotic myeloma cells exhibit unique characteristics, including enrichment of the cell-bound albumin and overexpression of endoplasmic reticulum (ER)- and mitochondrial-specific chaperones, which were not observed in other necrotic cells, including HUH-7, A2780, A549, and Hoc1a. Proteomic analysis shows that HSP60 is the most abundant up-regulated mitochondrial specific chaperone, and azacytidine-induced overexpression of HSP60 is confirmed by western blot analysis. In contrast, expression levels of cytosolic chaperones such as HSP90 and HSP71 were down-regulated in azacytidine-treated myeloma cells, concomitant with an increase of these chaperones in the cell culture medium, suggesting that mitochondrial chaperones and cytosolic chaperones behave differently in necrotic myeloma cells; ER- and mitochondrial-chaperones being retained, and cytosolic chaperones being released into the cell culture medium through the ruptured cell membrane. Our data suggest that HSP60 is potentially a new target for multiple myeloma chemotherapy.