Proteomic Analysis of Hylocereus polyrhizus Reveals Metabolic Pathway Changes.

Red dragon fruit or red pitaya (Hylocereus polyrhizus) is the only edible fruit that contains betalains. The color of betalains ranges from red and violet to yellow in plants. Betalains may also serve as an important component of health-promoting and disease-preventing functional food. Currently, the biosynthetic and regulatory pathways for betalain production remain to be fully deciphered. In this study, isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analyses were used to reveal the molecular mechanism of betalain biosynthesis in H. polyrhizus fruits at white and red pulp stages, respectively. A total of 1946 proteins were identified as the differentially expressed between the two samples, and 936 of them were significantly highly expressed at the red pulp stage of H. polyrhizus. RNA-seq and iTRAQ analyses showed that some transcripts and proteins were positively correlated; they belonged to "phenylpropanoid biosynthesis", "tyrosine metabolism", "flavonoid biosynthesis", "ascorbate and aldarate metabolism", "betalains biosynthesis" and "anthocyanin biosynthesis". In betalains biosynthesis pathway, several proteins/enzymes such as polyphenol oxidase, CYP76AD3 and 4,5-dihydroxy-phenylalanine (DOPA) dioxygenase extradiol-like protein were identified. The present study provides a new insight into the molecular mechanism of the betalain biosynthesis at the posttranscriptional level.

[Application of lymphography in the location and treatment decision of chyle leakage: an analysis of 177 cases].

OBJECTIVE: To identify the value of lymphography in the location and treatment decision of chyle leakage. METHODS: The clinic data of 177 patients suffered from chyle leakage admitted in 6 medical centers in Shanghai from February 1998 to December 2014 was analyzed retrospectively. There were 94 male and 83 female patients aging from 9 to 84 years with a mean of 49 years, including 128 cases of chyluria, 34 cases of primary chylothorax and 15 cases of other chyle leakage. All patients had failed to conservative treatment more than 2 weeks. Pedal lymphography was performed in every patient to investigate the site and range of chyle leakage. Effect of surgical or conservative management was compared according to the different results of lymphography. RESULTS: No serious complication was noticed. For all 177 patients, lymphography showed localized lymphatic diseases in 148 cases (83.6%), including 125 cases of lymphatic renal pelvic leaks, 14 cases of unilateral identified leak within thorax and 9 cases of chyle leakage in neck, heart, abdomen or scrotum. Among these patients, surgical treatment cured 129 and improved 3 patients but failed in 2 patients, while the remaining 14 cases had their leaks decreased after lymphography and cured by conservative management. For those 15 patients having disseminated lymphatic diseases or 14 with no abnormality under lymphography, surgery only cured 2 and improved 1 patient but failed in 8 patients (with 3 death), whereas continuous conservative treatment cured 11 patients, improved 5 patients but only failed in 2 patients (with one death). For localized leakage, surgical treatment showed better efficacy (98.5% vs. 3/11), whereas conservative treatment had significantly higher successful rate than surgical interventions in patients with disseminated lymphatic diseases or no abnormality under lymphography (16/18 vs. 3/11). CONCLUSIONS: Lymphography could identify the location and range of complicate chyle leakage failed to primary conservative management. Patients with disseminated lymphatic diseases or no abnormality under lymphography would be better managed by continuous non-operative treatment partly due to therapeutic effect of lymphography, while surgical intervention could be a good option for patients having localized lymphatic etiology.

Antagonism of rhizosphere Trichoderma brevicompactum DTN19 against the pathogenic fungi causing corm rot in saffron (Crocus sativus L.) in vitro.

Introduction: Corm rot in saffron (Crocus sativus L.) significantly impacts yield and quality. Non-toxic fungi, particularly Trichoderma species, are valuable for biological control due to their production of diverse and biologically active secondary metabolites. Methods: This study aimed to isolate an effective antagonistic fungus against the pathogenic fungi causing corm rot in saffron. Four pathogenic fungi (Fusarium oxysporum, Fusarium solani, Penicillium citreosulfuratum, and Penicillium citrinum) were isolated from diseased saffron bulbs in Chongming. Initial screening through dual culture with these pathogens re-screening from rhizosphere soil samples of C. sativus based on its inhibitory effects through volatile, nonvolatile, and fermentation broth metabolites. The inhibitory effect of biocontrol fungi on pathogenic fungi in vitro was evaluated by morphological observation and molecular biology methods. Results: Antagonistic fungi were identified as Trichoderma brevicompactum DTN19. F. oxysporum was identified as the most severe pathogen. SEM (scanning electron microscope) and TEM (transmission electron microscope) observations revealed that T. brevicompactum DTN19 significantly inhibited the growth and development of F. oxysporum mycelium, disrupting its physiological structure and spore formation. Additionally, T. brevicompactum DTN19 demonstrated nitrogen fixation and production of cellulase, IAA (Indole acetic acid), and siderophores. Whole-genome sequencing of strain DTN19 revealed genes encoding protease, cellulase, chitinase, ß-glucosidase, siderophore, nitrogen cycle, and sulfate transporter-related proteins. Discussion: T. brevicompactum DTN19 may inhibit the propagation of pathogenic fungi by destroying their cell walls or producing antibiotics. It can also produce IAA and iron carriers, which have the potential to promote plant growth. Overall, T. brevicompactum DTN19 showed the development prospect of biological agents.

Non-classical action of Ku70 promotes Treg suppressive function through a FOXP3-dependent mechanism in lung adenocarcinoma.

Ku70, a DNA repair protein, binds to the damaged DNA ends and orchestrates the recruitment of other proteins to facilitate repair of DNA double-strand breaks. Besides its essential role in DNA repair, several studies have highlighted non-classical functions of Ku70 in cellular processes. However, its function in immune homeostasis and anti-tumor immunity remains unknown. Here, we discovered a marked association between elevated Ku70 expression and unfavorable prognosis in lung adenocarcinoma, focusing specifically on increased Ku70 levels in tumor-infiltrated Treg cells. Using a lung-colonizing tumor model of in mice with Treg-specific Ku70 deficiency, we demonstrated that deletion of Ku70 in Treg cells led to a stronger anti-tumor response and slower tumor growth due to impaired immune-suppressive capacity of Treg cells. Furthermore, we confirmed that Ku70 played a critical role in sustaining the suppressive function of human Treg cells. We found that Ku70 bound to FOXP3 and occupied FOXP3-bound genomic sites to support its transcriptional activities. These findings not only unveil a non-homologous end joining (NHEJ)-independent role of Ku70 crucial for Treg suppressive function, but also underscore the potential of targeting Ku70 as an effective strategy in cancer therapy, aiming to both restrain cancer cells and enhance pulmonary anti-tumor immunity.

ULK1-mediated metabolic reprogramming regulates Vps34 lipid kinase activity by its lactylation.

Autophagy and glycolysis are highly conserved biological processes involved in both physiological and pathological cellular programs, but the interplay between these processes is poorly understood. Here, we show that the glycolytic enzyme lactate dehydrogenase A (LDHA) is activated upon UNC-51-like kinase 1 (ULK1) activation under nutrient deprivation. Specifically, ULK1 directly interacts with LDHA, phosphorylates serine-196 when nutrients are scarce and promotes lactate production. Lactate connects autophagy and glycolysis through Vps34 lactylation (at lysine-356 and lysine-781), which is mediated by the acyltransferase KAT5/TIP60. Vps34 lactylation enhances the association of Vps34 with Beclin1, Atg14L, and UVRAG, and then increases Vps34 lipid kinase activity. Vps34 lactylation promotes autophagic flux and endolysosomal trafficking. Vps34 lactylation in skeletal muscle during intense exercise maintains muscle cell homeostasis and correlates with cancer progress by inducing cell autophagy. Together, our findings describe autophagy regulation mechanism and then integrate cell autophagy and glycolysis.

Comparison of outcomes following lobectomy, segmentectomy, and wedge resection based on pathological subtyping in patients with pN0 invasive lung adenocarcinoma ≤1 cm.

PURPOSE: We sought to analyze the prognostic significance of lung adenocarcinoma classification for patients with pathological N0 (pN0) lung invasive adenocarcinomas ≤1 cm who underwent surgical resection and investigate the optimal surgical procedure according to lung adenocarcinoma classification. METHODS: A total of 1409 consecutive patients with resected pN0 invasive lung adenocarcinoma ≤1 cm were retrospectively reviewed. Comprehensive histologic subtyping was determined according to IASLC/ATS/ERS lung adenocarcinoma classification. Recurrence-free survival (RFS) and overall survival (OS) were compared between patients receiving lobectomy, segmentectomy, and wedge resection. RESULTS: RFS and OS favored lobectomy and segmentectomy compared with wedge resection in the entire cohort. Five-year RFS rates were 100%, 98.2%, 97.3%, 77.8%, and 82.8% (p < 0.001) for lepidic, acinar, papillary, micropapillary, and solid predominant subtypes, while 5-year OS rates were 100%, 98.4%, 98.1%, 88.9%, and 96.5% (p < 0.001), respectively. Multivariate analysis showed that adenocarcinoma predominant pathological subtype and CT appearance were independent prognostic factors for RFS, and surgical procedure was independent factor for both RFS and OS. Specifically, wedge resection showed worse survival compared with anatomical resection in patients with papillary, micropapillary, or solid predominant subtypes, whereas in patients with lepidic predominant and acinar predominant subtypes, wedge resection showed comparable RFS with anatomical resection. CONCLUSIONS: Anatomical resection showed better survival for patients with pN0 invasive lung adenocarcinoma ≤1 cm. For patients with invasive adenocarcinoma ≤1 cm in whom anatomical resection is not feasible, wedge resection could provide similar oncological effect when tumor is lepidic predominant or acinar predominant.

Surgical site infection following minimally invasive lobectomy: Is robotic surgery superior?

BACKGROUND: Surgical site infection (SSI) in thoracic surgery remains a significant cause of morbidity and prolonged hospitalization. Minimally invasive surgery (MIS) has significantly reduced the risk of SSI. We intended to compare whether there was difference between video-assisted thoracic surgery (VATS) and robotic-assisted thoracic surgery (RATS) in SSI and highlight possible factors influencing SSI in lobectomy. METHODS: This retrospective study analyzed patients who underwent minimally invasive lobectomy from January 2018 to December 2019. All patients' clinical characteristics and surgery-related information which may be related to the likelihood of SSI were recorded. RESULTS: A total of 1231 patients' records were reviewed with 806 VATS and 425 RATS. SSI was classified as deep or superficial SSI. Eighty-six (7.0%) patients were found to develop an SSI with 62 patients having deep infections and 24 had superficial infection. No statistical difference in the incidence rate and category of SSI was observed between patients undergoing VATS and RATS. CONCLUSIONS: There was no difference in the incidence of SSI between VATS and RATS lobectomy. Male gender, heavy smoking, uncontrolled diabetes mellitus, body mass index (BMI) > 27.9, more blood loss, and the higher National Healthcare Safety Network (NHSN) risk index score (1 or 2) were the independent risk factors of SSI following minimally invasive lobectomy, while male gender, uncontrolled diabetes mellitus, BMI > 27.9, more blood loss and the higher NHSN risk index score (1 or 2) were the main predictors of deep SSI.

The presence of lepidic and micropapillary/solid pathological patterns as minor components has prognostic value in patients with intermediate-grade invasive lung adenocarcinoma.

BACKGROUND: The acinar- and papillary-predominant histological subtypes are the most common types of invasive lung adenocarcinoma and are considered "intermediate-grade" carcinomas with heterogeneous prognosis. This study investigated the prognostic significance of the lepidic and micropapillary/solid pathological patterns as minor components in patients with intermediate-grade lung adenocarcinomas. METHODS: A total of 697 patients with pathological N0M0 acinar/papillary-predominant lung adenocarcinomas ≤3 cm in diameter, who underwent curative resection in our institution between June 1, 2014 and August 31, 2016, were retrospectively enrolled in this study. Acinar/papillary-predominant lung adenocarcinomas were classified into four subtypes according to the presence of the minor pathological components lepidic (Lep), micropapillary (MP), and solid (S). The subtypes were MP/S-Lep+, MP/S-Lep-, MP/S+Lep+, and MP/S+Lep-. The 5-year recurrence-free survival (RFS) and overall survival (OS) were recorded. Factors affecting survival were analyzed by Cox regression method. RESULTS: Among 697 intermediate-grade lung adenocarcinomas, the distribution of patients was as follows: MP/S-Lep+ type (n=314; 45.0%), MP/S-Lep- type (n=144; 20.7%), MP/S+Lep+ type (n=133; 19.1%), and MP/S+Lep- type (n=106; 15.2%). The 5-year RFS rates were 98.7%, 94.4%, 94.0%, and 81.9%, respectively (P<0.001). The 5-year OS rates were 98.4%, 94.4%, 96.6%, and 87.7%, respectively (P<0.001). Multivariate analysis revealed that the MP/S+Lep- subtype was an independent poor prognostic factor of both RFS and OS. CONCLUSIONS: Acinar/papillary-predominant adenocarcinoma is an "intermediate-grade" carcinoma that can be further classified into subtypes according to the presence of lepidic and micropapillary/solid pathological patterns with significantly different prognosis. This classification may be useful in evaluating the recurrence risk and guiding adjuvant therapies in patients with acinar/papillary-predominant stage I lung adenocarcinoma.

Nongated Computed Tomography Predicts Perioperative Cardiovascular Risk in Lung Cancer Surgery.

BACKGROUND: The coronary artery calcification score (CACS), a strong predictor of cardiovascular events and mortality, can be assessed by nongated chest computed tomography (CT). The study aimed to determine whether CACS based on nongated CT is predictive of perioperative cardiovascular events during intermediate-risk lung cancer surgery. METHODS: In this retrospective, single-center study, we used nongated CT images to evaluate CACS in 4491 patients with lung cancer who underwent intermediate-risk surgeries. Perioperative cardiovascular events were defined as in-hospital cardiac death, nonfatal myocardial infarction, heart failure, atrial and ventricular arrhythmia with hemodynamic compromise, and complete heart block. Risk factors of perioperative cardiovascular events were identified by multivariate logistic regression analysis. RESULTS: In total, 110 inpatients (2.5%) experienced perioperative cardiac events. Coronary calcification was observed on nongated CT in 1070 (23.8%) patients. CACS was significantly associated with the incidence of cardiovascular events and longer hospital stays. According to receiver operating characteristic curve analysis, the CACS cutoff value was set to 1. In the multivariate analysis, CACS ≥1 (odds ratio, 1.75; 95% CI, 1.14-2.68; P = .011) or the number of calcified vessels (odds ratio, 1.23; 95% CI, 1.01-1.50; P = .043), age, forced expiratory volume in 1 second/predicted, operation time, and thoracotomy were predictive of cardiovascular complications. CONCLUSIONS: CACS is an independent predictor of severe perioperative cardiovascular risk in patients undergoing intermediate-risk lung cancer surgery. CACS may represent a valuable tool for preoperative risk assessment of these patients.

Remote monitoring of patient recovery following lung cancer surgery: a messenger application approach.

BACKGROUND: Repeated assessment of patient recovery after discharge is challenging. This study used a popular messenger application to remotely collect patient self-reported symptoms and their severity so as to monitor patient recovery and identify the factors affecting the recovery of symptoms following lung cancer surgery. METHODS: This prospective observational study was conducted at a single tertiary lung cancer center in China between November 2018 and June 2019. Participants received demonstration videos and repeated symptom surveys regarding pain and cough severity (assessed using numeric rating scores of 0-10 for pain and 0-6 for cough) at 2, 4, 6, 8, and 12 weeks after discharge via a smartphone program bound to the WeChat application. Patients who responded to at least 3 of the 5 post-discharge surveys were included in this study. The data were analyzed to investigate the symptom recovery and its related factors. RESULTS: Of the 826 patients enrolled, 589 (71.3%) responded to at least three surveys. The average pain score reduced from 4.1±2.5 at 2 weeks to 2.2±2.0 at 12 weeks (P<0.001). Factors associated with higher pain severity included the female gender, age over 60 years, thoracotomy, longer operation time (>90 minutes), and prolonged chest tube drainage (>7 days). The average cough score decreased from 2.34±1.30 at 2 weeks to 1.93±1.26 at 12 weeks (P<0.001). Being female and a prolonged operation time (>90 min) were related to increased cough severity. Sublobar resection and limited lymphadenectomy may contribute to lower cough severity post-surgery. CONCLUSIONS: The messenger application-based remote monitoring successfully collected post-discharge symptom information and identified factors associated with recovery following lung surgery.

Comparison of perioperative outcomes of robotic-assisted versus video-assisted thoracoscopic right upper lobectomy in non-small cell lung cancer.

BACKGROUND: Robotic-assisted thoracic surgery (RATS) has been widely used in the treatment of lung cancer. The perioperative outcomes of right upper lobectomy (RUL) using RATS and video-assisted thoracic surgery (VATS) were retrospectively investigated and compared. We aimed to summarize a single-center experience of RATS and 4-port unidirectional VATS in RUL, and to discuss the safety and the essentials of the surgery. METHODS: We retrospectively analyzed the 685 with non-small cell lung cancer (NSCLC) patients who underwent minimally invasive RUL in our center by the same surgical group from January 2015 to December 2019. Both RATS and VATS were performed with three ports with utility incision. The 685 participants were divided into RATS (335 cases) and VATS (350 cases) groups according to surgical method. Baseline characteristics and perioperative outcomes including dissected lymph nodes, postoperative duration of drainage, postoperative hospital stay, and incidence of postoperative complications were compared between the groups. RESULTS: In the 685 patients enrolled, the baseline characteristics were comparable, and no postoperative 30-day mortality or intraoperative blood transfusion were observed. Compared with VATS, RATS had less surgical duration (90.22±12.16 vs. 92.68±12.26 min, P<0.001), less length of stay (4.71±1.37 vs. 5.26±1.56 days, P<0.001), and decreased postoperative duration of drainage (3.49±1.15 vs. 4.09±1.57 days, P<0.001). No significant difference was observed in the lymph nodes dissection, blood loss, conversion rate and morbidities. The cost of RATS was much higher than VATS (85,329.41±12,893.44 vs. 68,733.43±14,781.32 CNY, P<0.001). CONCLUSIONS: Robot assisted RUL had similar perioperative outcomes compared to VATS RUL lobectomy using similar three port with utility incision technique. The advantages of RATS included finer dissection of lymph node, relatively less operation time, earlier chest tube removal and discharge.

Operative outcomes and long-term survival of robotic-assisted segmentectomy for stage IA lung cancer compared with video-assisted thoracoscopic segmentectomy.

BACKGROUND: Robotic anatomic segmentectomy (RATS) for early-stage lung cancer is being increasingly performed in spite of limited published evidence. To evaluate its safety and oncologic efficacy, we compared the outcomes of both RATS and video-assisted thoracoscopic (VATS) segmentectomy in patients with small-sized (<2 cm) peripheral stage IA lung cancer. METHODS: From November 2011 to January 2018, a total of 130 patients with resected stage IA non-small cell lung cancer (NSCLC) who underwent RATS (n=50) and VATS (n=80) pulmonary segmentectomy were included. Clinicopathologic data, recurrence rate, and survival were recorded. RESULTS: The demographics, pulmonary function, comorbidity, and tumor size were similar between RATS segmentectomy and VATS segmentectomy. The surgery time, intensive care unit stay, hospital stay, and blood loss were reduced in the RATS group compared to the VATS group. The number of totally dissected lymph nodes and postoperative complications were similar between the 2 groups. There was no operative mortality. The intensity of narcotic use during hospital stay and the time to return to routine daily activities were also reduced in the RATS group. There was no recurrence observed in the RATS group during the median 38-month follow-up period; meanwhile, during a median 85-month follow-up period in the VATS group, local recurrence and distant recurrence was observed in 2 patients (2.5%) and 3 patients (3.75%) respectively. There was no significant difference in the 5-year recurrence-free survival between the RATS and VATS groups (100% vs. 93.75%; P>0.05). CONCLUSIONS: RATS can be performed safely and effectively in patients with early-stage NSCLC. The reduced narcotic use and earlier return to routine daily activities of RATS patients might reflect its less traumatic nature as compared to VATS. For stage IA disease with small tumors (<2 cm), segmentectomy performed by RATS has better oncologic efficacy when compared to VATS, although in this study, this difference did not reach statistical difference.

The deubiquitinating enzyme UCHL1 promotes resistance to pemetrexed in non-small cell lung cancer by upregulating thymidylate synthase.

Rationale: Resistance to pemetrexed (PEM)-based chemotherapy is a major cause of progression in non-small cell lung cancer (NSCLC) patients. The deubiquitinating enzyme UCHL1 was recently found to play important roles in chemoresistance and tumor progression. However, the potential roles and mechanisms of UCHL1 in PEM resistance remain unclear. Methods: Bioinformatics analyses and immunohistochemistry were used to evaluate UCHL1 expression in NSCLC specimens. Kaplan-Meier analysis with the log-rank test was used for survival analyses. We established PEM-resistant NSCLC cell lines by exposing them to step-wise increases in PEM concentrations, and in vitro and in vivo assays were used to explore the roles and mechanisms of UCHL1 in PEM resistance using the NSCLC cells. Results: In chemoresistant tumors from NSCLC patients, UCHL1 was highly expressed and elevated UCHL1 expression was strongly associated with poor outcomes. Furthermore, UCHL1 expression was significantly upregulated in PEM-resistant NSCLC cells, while genetic silencing or inhibiting UCHL1 suppressed resistance to PEM and other drugs in NSCLC cells. Mechanistically, UCHL1 promoted PEM resistance in NSCLC by upregulating the expression of thymidylate synthase (TS), based on reduced TS expression after UCHL1 inhibition and re-emergence of PEM resistance upon TS restoration. Furthermore, UCHL1 upregulated TS expression, which mitigated PEM-induced DNA damage and cell cycle arrest in NSCLC cells, and also conferred resistance to PEM and other drugs. Conclusions: It appears that UCHL1 promotes PEM resistance by upregulating TS in NSCLC cells, which mitigated DNA damage and cell cycle arrest. Thus, UCHL1 may be a therapeutic target for overcoming PEM resistance in NSCLC patients.

Perioperative outcomes of radical lobectomies using robotic-assisted thoracoscopic technique vs. video-assisted thoracoscopic technique: retrospective study of 1,075 consecutive p-stage I non-small cell lung cancer cases.

BACKGROUND: Robotic thoracoscopic surgery was first done in mainland China in 2009 and has gained popularity in the past few years. Here, we present the largest Chinese series of robotic lobectomy for early-stage non-small cell lung cancer (NSCLC) to date. We aimed to compare the perioperative outcomes of our three-arm robotic-assisted lobectomy (RAL3) and video-assisted lobectomy (VAL) for p-stage I NSCLC and report the approach of the robotic anatomic lobar resections of our center. METHODS: We retrospectively collected and analyzed the data of 1075 stage I NSCLC patients who underwent minimally invasive lobectomies (237 RAL3 cases and 838 VAL cases) by the same surgical team from May 2013 to April 2016. Propensity score matching (PSM) was used to minimize the bias between the two groups. Perioperative outcomes were analyzed. RESULTS: Compared to the VALs, the RAL3s had more retrieved lymph nodes (LNs) (9.70 vs. 8.45, P=0.000), less POD1 drain (230.91 vs. 279.79 mL, P=0.001), shorter chest tube duration (3.84 vs. 4.33 d, P=0.003) and shorter postoperative length of stay (4.97 vs. 5.45 d, P=0.004), but a higher cost (¥93,244.84 vs. ¥67,055.82, P=0.000). No significant difference was observed between the RAL3 and VAL groups concerning the average skin-to-skin time (90.84 vs. 92.25 min, P=0.624), conversion rate (1.3% vs. 0.87%, P=1.000) and prolonged postoperative hospital stay (PPHS) rate (3.0% vs. 4.3%, P=0.694). CONCLUSIONS: This study confirms that RAL3 is a safer and more effective technique than VAL for the treatment of early-stage NSCLC.

Fatty acid receptor GPR120 promotes breast cancer chemoresistance by upregulating ABC transporters expression and fatty acid synthesis.

BACKGROUND: Chemoresistance is the major cause of neoadjuvant treatment failure in breast cancer patients. Despite recent progress, the mechanism underlying chemoresistance remains to be further defined. METHODS: Expression of G protein-coupled receptor 120 (GPR120) was analyzed by immunohistochemistry in the biopsies of primary breast cancer who subsequently underwent preoperative neoadjuvant chemotherapy. In vitro and in vivo loss- and gain-of -function studies were performed to reveal the effects and related mechanism of GPR120 signaling pathway in the chemoresistance of breast cancer cells. FINDINGS: We identified that GPR120, a receptor for long-chain fatty acids, was important for the acquisition of chemoresistance in breast cancer cells. We showed that GPR120 expression was positively associated with clinical response to neoadjuvant chemotherapy in patients. In breast cancer cells, GPR120 enhanced the de novo synthesis of fatty acids that served as GPR120 ligands to activate GPR120 signaling via a feedback mechanism. Upregulated GPR120 signaling rendered cells resistant to epirubicin-induced cell death by upregulating ABC transporters expression and thus decreasing the intracellular accumulation of epirubicin. Akt/NF-κB pathway was responsible for the GPR120-mediated expression of ABC transporters leading to modulation of the concentration of chemotherapeutic drugs in cells. The functional importance of GPR120 in chemoresistance was further validated using epirubicin-treated tumor xenografts, in which we showed that blockade of GPR120 signaling with AH7614 or GPR120-siRNA significantly compromised chemoresistance. INTERPRETATION: Our results highlight that GPR120 might be a promising therapeutic target for breast cancer chemoresistance. FUND: National Natural Science Foundation of China, Ministry of Science and Technology of China, Program of Science and Technology Commission of Shanghai Municipality.

Value of folate receptor-positive circulating tumour cells in the clinical management of indeterminate lung nodules: A non-invasive biomarker for predicting malignancy and tumour invasiveness.

BACKGROUND: Non-invasive lung adenocarcinoma could benefit from limited resection, nonetheless, there is a lack of method to determine preoperative tumour invasiveness. We aimed to investigate whether folate receptor-positive circulating tumour cells (FR+-CTCs) in combination with maximum tumour diameter (MTD) determines, before surgery, the invasiveness of small-sized, indeterminate solitary pulmonary nodules (SPNs). METHODS: A total of 382 patients with suspicious lung adenocarcinoma on computed tomography who were expected to undergo lung resection were enrolled in this study at three participating institutes and randomly assigned into training and validation cohorts. Before surgery, 3 mL peripheral blood was collected from all participants. FR+-CTCs were analyzed using immunomagnetic leukocyte depletion and quantitated by ligand-targeted PCR method. After surgery, the resected tissues were diagnosed by pathologists according to IASLC/ATS/ERS classification. FINDINGS: FR+-CTC levels in the peripheral blood can differentiate benign from malignant nodules with a sensitivity of 78·6%-82·7% and a specificity of 68·8%-78·4%. Both FR+-CTC and MTD are independent predictive indices of invasive tumours for lung adenocarcinoma ≤2 cm based on multivariate analyses. Further, FR+-CTC count in combination with MTD can differentiate non-invasive cancers from invasive cancers with a sensitivity of 63·6%-81·8% and a specificity of 71·4%-89·7%. INTERPRETATION: Detection of FR+-CTC is a reliable method to differentiate malignancy of indeterminate SPNs. Combining of FR+-CTC count and MTD could possibly enhance preoperative determination of the invasiveness of lung nodules and guide surgeons to select limited lung resection and avoid overtreatment for patients with non-invasive lesions. FUND: None.

Aberrant fatty acid profile and FFAR4 signaling confer endocrine resistance in breast cancer.

BACKGROUND: Evidence suggests that fatty acid receptor FFAR4 plays a tumor-promoting role in adipose tissue-adjacent malignancies, but its clinical relevance remains unexplored. Here, we investigated the clinical significance and underlying mechanisms of FFAR4 in hormone receptor-positive breast cancer (HRPBC). METHODS: FFAR4 expression was assessed by immunohistochemistry in an exploration cohort of 307 breast cancer cases collected from two independent institutes. Two public breast cancer microarray datasets served as validation cohorts. Gas chromatography-mass spectrometry was employed to identify FFAR4 ligands in normal and cancerous breast tissues. Survival analyses were performed in all cohorts and designated molecular subgroups. Mechanistic studies were performed in vitro in hormone receptor-positive breast cancer cell lines MCF-7 and T-47D. RESULTS: Aberrant FFAR4 expression and endogenous FFAR4 ligands were identified in breast cancer tissues, five FFAR4 ligands showed significantly elevated proportions in cancerous versus normal tissues. In the exploration cohort, FFAR4 was demonstrated as an independent prognostic factor for recurrences (HR: 2.183, 95% CI: 1.360-3.504, P = 0.001) and breast cancer-specific deaths (HR: 2.102, 95% CI: 1.173-3.766, P = 0.013) in HRPBC cases. In contrast, FFAR4 expression was not associated with prognosis in hormone receptor-negative cases. In the validation cohorts, FFAR4 mRNA levels were also observed to be associated with disease recurrence in estrogen receptor-positive cases, but not so in estrogen receptor-negative cases. FFAR4 activation by endogenous ligands and a synthetic ligand TUG891 significantly dampened tamoxifen's efficacy on HRPBC cells, whereas FFAR4 knockdown or antagonist AH7614 abrogated this effect. Furthermore, FFAR4-induced tamoxifen resistance was dependent on ERK and AKT pathways in HRPBC. CONCLUSIONS: Our results establish a novel role of FFAR4 and its ligands in the complicated interactions between tissue lipid profile and cancer biology. FFAR4 signaling confers tamoxifen resistance in HRPBC cell line and FFAR4 expression can serve as a prognostic biomarker for tamoxifen-treated HRPBC patients. FFAR4 may serve as a potential target for anti-breast cancer therapies, especially in endocrine resistant cases.

Clinical implication of the expression of Aurora B in normal endometrium and endometrial carcinoma.

The expression of Aurora B in normal endometria and endometrial carcinomas and its relation with clinicopathologic parameters of endometrial carcinomas were investigated. Streptavidin-biotin peroxidase (SP) immunohistochemical technique was used to detect the expression of Aurora B in 10 cases of normal proliferative phase endometria, 10 cases of normal secretory phase endometria and 72 cases of endometrial carcinomas respectively. According to the 1988 International Federation of Gynecology and Obstetrics (FIGO) grade, there were 37 patients in grade 1, 23 in grade 2 and 12 in grade 3 respectively. According to the FIGO stage, there were 59 patients in stage I-II and 13 patients in stage III-IV. Aurora B was expressed in both normal proliferative phase endometria, secretory phase endometria and endometrial carcinomas, but its positive labeling index (PLI) in proliferative phase endometria was significantly higher than that in secretory phase endometria (P<0.01) and endometrial carcinomas (P<0.01). The PLI of Aurora B was lower in tumors with well differentiation (G(1)), low surgical staging (I-II), and 1/2 myometrial invasion (all P<0.01). Aurora B exerts its functions in the replication of normal endometrial glandular cells; Expression of Aurora B is significantly correlated with biologic behavior of endometrial carcinoma, indicating that Aurora B may be a promising prognostic factor in endometrial carcinoma.

Gambogenic acid inhibits fibroblast growth factor receptor signaling pathway in erlotinib-resistant non-small-cell lung cancer and suppresses patient-derived xenograft growth.

Erlotinib resistance causes a high degree of lethality in non-small-cell lung cancer (NSCLC) patients. The high expression and activation of several receptor tyrosine kinases, such as JAK/STAT3, c-Met, and EGFR, play important roles in drug resistance. The development of tyrosine kinase inhibitors is urgently required in the clinic. Our previous study found that Gambogenic acid (GNA), a small molecule derived from the traditional Chinese medicine herb gamboge, induced cell death in several NSCLC cell lines through JAK/STAT3 inhibition. In this study, we investigated the mechanism of action of GNA in erlotinib-resistant NSCLC and patient-derived cells. The inhibition of GNA on FGFR signaling pathway was examined using biochemical kinase assays. NSCLC cell lines (HCC827, HCC827-Erlotinib-resistant, and H1650) and primary cells from patients with NSCLC with clinical resistance to erlotinib were treated with GNA, erlotinib, or their combination. Both kinase assays and cell- based assays showed that GNA inhibits the phosphorylation of multiple kinases in FGFR signaling pathway in NSCLC. The combination of GNA and erlotinib significantly attenuates the tumor growth of HCC827 and erlotinib-resistant HCC827 xenografts with low toxicity. Importantly, GNA significantly suppresses tumor growth in a lung patient-derived xenograft (PDX) model with FGFR fusion and low EGFR expression. Our findings provide preclinical evidence for using GNA as an FGFR signaling pathway inhibitor to overcome erlotinib resistance in NSCLC treatment or to enhance erlotinib efficacy when used as a combined administration.

Targeting the NRF-2/RHOA/ROCK signaling pathway with a novel aziridonin, YD0514, to suppress breast cancer progression and lung metastasis.

Metastasis is a major cause of breast cancer-associated mortality. Natural products extracted from herbs provide rich bioactive compounds with anticancer efficacy but may have limited or moderate potency and considerable toxicity. We developed a novel aziridonin, YD0514, by aziridinating oridonin, a natural product of the medicinal herb Rabdosia rubescens. In this study, we found that YD0514 significantly inhibited proliferation, motility, and adhesion of metastatic breast cancer cell lines MDA-MB-231, GI101, GILM2, and GILM3. YD0514 also decreased the protein expression of matrix metalloproteinases 2 and 9 (MMP2 and MMP9), focal adhesion kinase (FAK), and integrin family members. Importantly, YD0514 suppressed the growth of metastatic breast cancer xenograft tumors and significantly inhibited lung metastasis in vivo. Lastly, we showed that YD0514's anti-metastatic effect on highly aggressive breast cancer is mediated via regulating the NRF-2/RHOA/ROCK signaling pathway. These results demonstrate that YD0514, the first active analog based on an oridonin D-ring modification, has the potential to be developed as an anti-metastasis therapy for patients with metastatic cancers.