Understanding the functions and regulatory mechanisms of the epitranscriptome entails robust and accurate analytical methods to identify and quantify post-transcriptional modifications in RNA. However, there are still various challenges in analyzing multiple modified nucleosides in RNA. Herein, we established a highly sensitive and high-throughput hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method, in conjunction with a stable isotope-dilution technique, for accurate quantification of 35 nucleosides. By the use of malic acid as a mobile phase additive, the HILIC-based separation of nucleosides was improved and the MS signal response of nucleosides was enhanced by 2.5- to 20.0-fold. Notably, seven groups of isomeric nucleosides with identical multiple-reaction monitoring ion transitions and six groups of nucleosides with identical or similar molecular weights that were indistinguishable by MS were well resolved by optimal HILIC separation. Thirty-five nucleosides were analyzed simultaneously within 12.5 min, and the limits of detection of these nucleosides ranged from 15.0 amol to 43.5 fmol. With this method, we conducted a comprehensive analysis and evaluation of the alteration in the RNA modification profile in breast cancer and assessed the RNA modification patterns across different breast cancer subtypes. The developed HILIC-MS/MS method has excellent capabilities for sensitive and high-throughput detection of multiple modified nucleosides, thereby providing a valuable analytical tool for deciphering the epitranscriptomic landscape and screening nucleosides as biomarkers in future clinical research.

1050 Background: Breast cancer brain metastasis(BCBM) is a major cause of mortality in advanced breast cancer patients, and treatment options are limited. In this project, we investigated the proteomic landscape of primary breast cancer and brain metastasis. Methods: We conducted high-throughput proteomic analysis on primary and brain metastatic breast cancer tissues surgically resected from 21 patients. Following screening and sample processing using pressure cycle technology (PCT) for peptide extraction, we used the data-independent acquisition (DIA) mass spectrometry (MS) method to acquire the data. Additionally, we used biological function assays, co-culture experiments, and transcriptome sequencing analysis to further investigate the differentially expressed proteins. Results: Patients were mostly in clinical stage II, with two excluded from analysis due to no surgery. Breast cancer classifications included 47.6% HER2+/HR-, 28.6% HER2+/HR+, 9% HER2-/HR+, and 14.3% TNBC. All patients received chemotherapy, with 57% undergoing neoadjuvant therapy and 12 receiving targeted therapy (all with trastuzumab). A total of 9430 protein groups were identified, with 692 showing differential expression in BCBM. Notably upregulated proteins included CRYAB, GFAP, STXBP1 and significantly downregulated proteins included CACNA1A, AOC3, PMP2, OGN. Most differentially expressed proteins were involved in extracellular matrix (ECM) and cell-cell interactions, with collagen family proteins (e.g., COL14A1, COL22A1) playing key roles in BCBM. HER2+ BCBM was associated with ECM pathways, while TNBC impacted the immune microenvironment. Regardless of the subtype, we identified 11 proteins that collectively contribute to the development of BCBM, including CRYAB, ATP6V0A1, HLA-DQB1, TPM2, SERPINB9, NFATC2, GRAP, ALDH1L2, DHRS4L2, SEPTIN1 and SAMHD1. We found that high ACOX1 and low KRT9/KRT14 expression were linked to poor prognosis in BCBM. Analyzing whether patients had undergone targeted therapy, we found that resistance might be acquired through the oxidative phosphorylation pathway, promoting brain metastasis in HER2+ breast cancer. Interestingly, CRYAB expression was prominent across all subtypes of BCBM and targeted therapy groups, suggesting it may serve as an essential biomarker for BCBM. We have confirmed that CRYAB can promote the proliferation and migration of HER2+BCBM, and preliminarily verified that CRYAB is closely related to immune infiltration through CXCL5, CXCL8 and CCL3 in tumor microenvironment, which may promote the occurrence of HER2+BCBM by affecting the NF-κB pathway. Conclusions: Leveraging high-throughput proteomics, we present a detailed analysis that elucidates the biological processes involved in developing and progressing BCBM from multiple angles. This work offers new directions for early prediction, treatment, and prognosis of BCBM in clinical practice.

Breast cancer, the most prevalent malignant tumor in women, is closely linked to the human microbiota. The microbiome participates throughout breast cancer pathogenesis, including its occurrence, progression, response to anti-tumor therapies, and treatment-related complications. This review examines the central hypothesis that microbiome-driven inflammatory and immune mechanisms shape breast cancer progression through two key pathways: systemic immune-inflammatory regulation and local tumor microenvironment remodeling. Furthermore, microorganisms and their metabolites modulate systemic treatments by interfering with drug metabolism and altering systemic or local immune-inflammatory environments. Targeting the microbiota represents a promising strategy for enhancing anticancer efficacy and reducing treatment-related complications. This review aims to advance the understanding of the etiology and disease progression of breast cancer from the perspective of microbial-regulated inflammation and immunity, offering new insights for its prevention and treatment.

RNA modifications play vital regulatory roles in biological systems. Dysregulated RNA modifications themselves or their regulators are associated with various diseases, including cancers and immune related diseases. However, to the best of our knowledge, RNA modifications in peripheral white blood cells (immune cells) haven't been systematically investigated before. Here we utilized hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) for the quantification of 19 chemical modifications in total RNA and 17 chemical modifications in small RNA in peripheral white blood cells from breast cancer patients and healthy controls. We found out 13 RNA modifications were up-regulated in total RNA samples of breast cancer patients. For small RNA samples, only N6-methyladenosine (m6A) was down-regulated in breast cancer patients (P < 0.0001). Receiver operating characteristic (ROC) curves analysis showed that N4-acetylcytidine (ac4C) in total RNA had an area under curve (AUC) value of 0.833, and m6A in small RNA had an AUC value of 0.994. Our results further illustrated that RNA modifications may play vital roles in immune cell biology of breast cancer, and may act as novel biomarkers for the diagnosis of breast cancer.

Abstract Purpose The totally implanted venous access port (TIVAP) is widely applied to chemotherapy. The traditional approach is to implant the port by directly cutting skin at the chest wall, but surgical scarring on the chest wall may bring permanent psychological trauma to patients and affect the port once the incision is infected. In this study, the effect and safety of an improved port implantation (noninvasive chest wall implantation) via the axillary approach on breast cancer patients were evaluated. Methods This study discusses the surgical steps of the improved port implantation. The incidences of complication, comfort, convenience, aesthetics, and privacy from the improved operation were analyzed and compared with the traditional operation. Results All patients successfully presented improved infusion port implantation through the axillary access (noninvasive chest wall implantation). Two cases had a hemorrhage during the operation. One case had a postoperative subcutaneous hemorrhage, and one case had a folded catheter. Nevertheless, the patients did not need a secondary operation for adjustment. The average operation time of the improved infusion port implantation was 51.85 min (range: 37–69 min). The improved operation was significantly better than the traditional operation in terms of aesthetics and privacy. In terms of comfort and convenience, the difference between the two operations was not significant. Conclusions This study described the specific steps and particular aspects of the improved operation. The effectiveness and safety of the improved operation were reported for the first time. The improved operation has been proven safe and reliable, and it entails only a few intraoperative and postoperative complications.

BACKGROUND Various animal models have been used to explore the pathogenesis of choledochal cysts (CCs), but with little convincing results. Current surgical techniques can achieve satisfactory outcomes for treatment of CCs. Consequently, recent studies have focused more on clinical issues rather than basic research. Therefore, we need appropriate animal models to further basic research. AIM To establish an appropriate animal model that may contribute to the investigation of the pathogenesis of CCs. METHODS Eighty-four specific pathogen-free female Sprague-Dawley rats were randomly allocated to a surgical group, sham surgical group, or control group. A rat model of CC was established by partial ligation of the bile duct. The reliability of the model was confirmed by measurements of serum biochemical indices, morphology of common bile ducts of the rats as well as molecular biology experiments in rat and human tissues. RESULTS Dilation classified as mild (diameter, ≥ 1 mm to < 3 mm), moderate (≥ 3 mm to < 10 mm), and severe (≥ 10 mm) was observed in 17, 17, and 2 rats in the surgical group, respectively, while no dilation was observed in the control and sham surgical groups. Serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, direct bilirubin, and total bile acids were significantly elevated in the surgical group as compared to the control group 7 d after surgery, while direct bilirubin, total bilirubin, and gamma-glutamyltransferase were further increased 14 d after surgery. Most of the biochemical indices gradually decreased to normal ranges 28 d after surgery. The protein expression trend of signal transducer and activator of transcription 3 in rat model was consistent with the human CC tissues. CONCLUSION The model of partial ligation of the bile duct of juvenile rats could morphologically simulate the cystic or fusiform CC, which may contribute to investigating the pathogenesis of CC.

PURPOSE:This study was a retrospective and nonrandomized study to assess the safety and reliability of identifying the surgical margin in breast cancer breast-conserving surgery (BCS) by using intraoperative ultrasonic location and specimen mammography instead of traditional intraoperative frozen pathological section.METHODS:Among the patients who underwent BCS from May 2019 to October 2021, according to the different methods of evaluating the intraoperative margin, 104 breast cancer patients were included in the frozen edge group, 53 breast cancer patients were included in the freeze-free group, and the surgeon judged whether extended resection was needed based on the results of pathological section or evaluation of intraoperative ultrasound and mammography. The surgical margins of the two groups were judged by postoperative pathological results as the gold standard.RESULTS:The median waiting pathology results time in the frozen edge group was 64 minutes, while the waiting time in the freeze-free group was 30 minutes, and the difference was statistically significant (P < .0001). The postoperative pathological results showed that the positive rate of the surgical margin in the frozen edge group was 0.96%. The coincidence rate of intraoperative frozen and postoperative pathological results was 99.04%. The coincidence rate between intraoperative mammography and postoperative pathological results was 100%.CONCLUSIONS:In BCS, the method of using intraoperative staining markers combined with mammography to evaluate the resection margin is highly accurate, reliable, economical and convenient, and at the same time reduces the waiting time of the operator during the operation. However, this was not a randomized controlled study, and there was patient selection bias, and its safety needs to be confirmed by long-term follow-up. In the future, it is expected to become the mainstream means of evaluating.

Breast cancer is the most frequent malignancy in women. However, in the middle and late stages, some people develop distant metastases, which considerably lower the quality of life and life expectancy. The brain is one of the sites where metastasis frequently happens. According to epidemiological research, brain metastases occur at a late stage in 30-50% of patients with HER2-positive breast cancer, resulting in a poor prognosis. Additionally, few treatments are available for HER2-positive brain metastatic breast cancer, and the mortality rate is remarkable owing to the complexity of the brain's anatomical structure and physiological function. In this review, we described the stages of the brain metastasis of breast cancer, the relationship between the microenvironment and metastatic cancer cells, and the unique molecular and cellular mechanisms. It involves cancer cells migrating, invading, and adhering to the brain; penetrating the blood-brain barrier; interacting with brain cells; and activating signal pathways once inside the brain. Finally, we reviewed current clinically used treatment approaches for brain metastasis in HER2-positive breast cancer; summarized the traditional treatment, targeted treatment, immunotherapy, and other treatment modalities; compared the benefits and drawbacks of each approach; discussed treatment challenges; and emphasized the importance of identifying potential targets to improve patient survival rates and comprehend brain metastasis in breast cancer.

The objective of this study is to assess the predictive potency of cell senescence-related genes (CSRGs) in breast cancer (BC) and establish a risk signature. Trascriptome data of CSRGs were obtained from the TCGA and GEO databases. Consensus clustering was used to generate CSRGs-based molecular clusters for BC patients. A CSRGs-derived risk signature was built using multiple Cox regression analyses of differentially expressed genes (DEGs) between clusters. The prognosis, immune infiltration, chemotherapy and immunotherapy response between different risk groups were analyzed and compared. Two molecular clusters of BC patients were generated on the basis of 79 differentially expressed CSRGs, which showed distinct prognosis and immune infiltration. A total of 1403 DEGs between the CSRGs-derived clusters were found, and 10 of them were independent prognostic genes that used to construct a risk signature. The results demonstrated that patients with older age and advanced stage presented with a higher risk scores. In addition, the risk signature was found to be associated with outcomes, immune infiltration, chemotherapy and immunotherapy response. Patients in the low-risk group showed a favorable prognosis and higher immunotherapy response than those in the high-risk group. Finally, we developed a highly stable nomogram that incorporates risk signature, chemotherapy, radiotherapy, and stage variables, enabling accurate prediction of the overall survival (OS) of individual patients. To conclude, the signature derived from CSRGs holds great promise as a biomarker for prognostic assessment of BC and may serve as a valuable tool in guiding immunotherapy.

BACKGROUND:There are few studies comparing robotic-assisted surgery (RAS) and laparoscopic-assisted surgery (LAS) in Hirschsprung's disease (HSCR). This study aimed to compare intraoperative and postoperative outcomes between RAS and LAS performed during the same period. METHODS:All consecutive 75 patients with pathologically diagnosed as HSCR who underwent Swenson pull-through surgery from April 2020 to Nov 2022, were included. Patients were divided into RAS group and LAS group and a retrospective analysis was performed based on clinical indexes and prognosis. RESULTS:A total of 75 patients were included, among which, 31 patients received RAS and 44 received LAS. The RAS and LAS groups had similar ages, sex, weight, postoperative hospital stays, and fasting times. Compared with LAS, blood loss (p = 0.002) and the incidence of Hirschsprung-associated enterocolitis (p = 0.046) were significantly lower in the RAS group. The first onset of Hirschsprung-associated enterocolitis in patients younger than 3 months occurred significantly earlier (p = 0.043). Two patients experienced anastomotic leakage in the LAS group and one patient experienced incisional hernia in the RAS group. The cost of RAS was significantly higher than that of LAS (p < 0.0001). CONCLUSIONS:RAS is a safe and effective alternative for HSCR children, and a delaying primary surgery until later in infancy (> 3 months) may improve outcomes.

BackgroundHepatoid adenocarcinoma of the stomach (HAS) is a highly malignant subtype of gastric carcinoma with specific clinicopathological features and extremely poor prognosis. We present an exceedingly rare case of complete response after chemo-immunotherapy.Case DescriptionA 48-year-old woman with highly elevated serum alpha-fetoprotein (AFP) level was found to have HAS verified by pathological examination based on gastroscopy. Computed tomography scan was done and TNM staging of the tumor was T4aN3aMx. Programmed cell death ligand-1 (PD-L1) immunohistochemistry was performed, revealing a negative PD-L1 expression. Chemo-immunotherapy including oxaliplatin plus S-1 and PD-1 inhibitor terelizumab was given to this patient for 2 months until the serum AFP level decreased from 748.5 to 12.9 ng/mL and the tumor shrank. D2 radical gastrectomy was then performed and histopathology of the resected specimen revealed that the cancerous cells had disappeared. Pathologic complete response (pCR) was achieved and no evidence of recurrence has been found after 1 year of follow-up.ConclusionsWe, for the first time, reported an HAS patient with negative PD-L1 expression who achieved pCR from the combined chemotherapy and immunotherapy. Although no consensus has been reached regarding the therapy, it might provide a potential effective management strategy for HAS patient.

Background: Peripheral blood inflammation indices, including the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and systemic immune-inflammation index (SII), have become research hotspots in the diagnosis, treatment, and prognosis prediction of breast cancer, whereas existing research findings remain controversial.Methods: Data pertaining to 1808 breast cancer patients were collected retrospectively to analyze the predictive value of NLR/PLR/ SII for breast cancer clinicopathological characteristics, chemotherapy response, and relapse. 1489, 258, and 53 eligible breast cancer patients entered into the three analyses, respectively. Logistic regression analyses were used to assess the correlation between these indices and poor response to chemotherapy. A predictive scoring model was established to predict chemotherapeutic responses based upon the odds ratio values of significant variables identified in logistic regression analyses. Results: Higher pretherapeutic NLR/PLR/SII values were significantly correlated with higher tumor stage, triple-negative breast cancer, premenopausal status, and younger age. Logistic regression analyses indicated that pretherapeutic high SII (as a continuous variable or with a cut-off value of 586.40) and HER2-negative status were independent predictors of poor response to neoadjuvant chemotherapy. A first-in-class SII-based predictive scoring model well distinguished patients who might not benefit from neoadjuvant chemotherapy, with an area under the curve of 0.751. In HR-positive cancers, SII was more strongly associated with clinicopathological features and chemotherapy response. In addition, a receiver operating characteristic curve analysis indicated that the specificity of follow-up SII in identifying cancer relapse was greater than 98.0% at a cut-off value of 900.Conclusion: As a predictor of breast cancer, especially in the HR-positive subtype, SII may eclipse NLR/PLR. SII-high patients are more likely to have a worse chemotherapy response and a higher risk of recurrence.

In breast conserving surgery (BCS), specimen mammography is one of the most widely used intraoperative methods of assessing margin status. We performed a meta-analysis to evaluate the diagnostic accuracy of specimen mammography. Literature databases including PubMed, Cochrane Library, Web of Science, and EMBASE were searched prior to Jun 2022. A total of 1967 patients were included from 20 studies. A pooled analysis, heterogeneity testing, threshold effect testing, publication bias analysis, and subgroup analyses were performed from extracted data. The pooled weighted values were a sensitivity of 0.55 (95% confidence interval [CI], 0.47–0.63), a specificity of 0.85 (95% CI, 0.78–0.90), a diagnostic odds ratio of 7 (95% CI, 4–12), and a pooled positive likelihood ratio of 3.7 (95% CI 2.6–5.5). The area under the receiver operator characteristic curve was 0.75 (95% CI 0.71–0.78). In the subgroup analysis, the pooled specificity in the positive margin defined as tumor at margin subgroup was lower than the other positive margin definition subgroup (0.82 [95% CI: 0.71, 0.92] vs. 0.87 [95% CI: 0.80, 0.94], p = 0.01). Our findings indicated that specimen mammography was an accurate intraoperative imaging technique for margin assessment in BCS.

RNA methylation plays a vital role in the pathogenesis of a variety of diseases including cancer, and aberrant levels of modified nucleosides in RNA were revealed to be related to cancer. Urine is a favored source for biomarker discovery due to the non-invasion to patients. Herein, we developed a sensitive hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC–MS/MS) method combined with stable isotope dilution for accurate quantification of methylated nucleosides in human urine. With this method, we successfully quantified ten methylated nucleosides in urine samples collected from healthy controls and breast cancer patients. We found N6-methyladenosine (m6A), 2′-O-methyladenosine (Am), N1-methyladenosine (m1A), N6,2′-O-dimethyladenosine (m6Am), N1-methylguanosine (m1G), 2′-O-methylguanosine (Gm), 5-methylcytidine (m5C) and 2′-O-methylcytidine (Cm) were all decreased in early-stage breast cancer patients, and a nomogram prediction model was constructed. Locally advanced breast cancer patients exhibited elevated levels of urinary 2′-O-methylated nucleosides in comparison to early-stage breast cancer patients. Together, we developed a robust method for the simultaneous determination of methylated nucleosides in human urine, and the results revealed an association between the contents of urinary methylated nucleosides and the occurrence of breast cancer, which may stimulate future studies about the regulatory roles of these methylated nucleosides in the initiation and progression of breast cancer.

Magnetic foreign bodies ingestion is a special cause for attending emergency department. Here, we aim to analyze the characteristics and treatments of children who ingested magnetic foreign bodies (Buckyballs). Data were collected from children who ingested Buckyballs between February 2017 and October 2019. A retrospective analysis was performed to summarize the experiences of conservative treatment, gastroscopy and surgery when dealing with Buckyballs ingestion. A total of 49 patients with buckyballs ingestion were identified, of whom 11 underwent conservative treatments, 6 underwent gastroscopy, and 32 underwent surgery. Among such individuals, eight patients (72.7%) had a successful conservative treatment (number of Buckyballs [NB]: 3.5[IQR: 2.0-4.0]); four patients (66.7%) had Buckyballs successfully removed by gastroscopy (NB: 3.5[IQR: 3.0-5.5]); 16 asymptomatic (50%) patients (NB: 4.0[IQR: 3.0-8.0]) and 16 symptomatic (50%) patients (NB: 8.5 [IQR: 6.25-11.75]) received emergency surgery. Compared to patients who received conservative treatment, the number of ingested Buckyballs was significantly higher in patients who received surgery or gastroscopy (7.0 [IQR: 3.0-10.75] vs 3.5 [IQR: 2.0-4.0], P < .05). The risk of intestinal perforation was significantly higher in symptomatic patients (P < .05) compared to asymptomatic patients. Gastroscopy is recommended when Buckyballs are in the stomach or esophagus. In asymptomatic patients, conservative treatment can be considered for 4 to 6 days. Patients failing conservative treatment, or those who are symptomatic should undergo emergency surgery.

Purpose . Triple-negative breast cancer (TNBC) is a heterogeneous and aggressive disease with poorer prognosis than other subtypes. We aimed to investigate the prognostic efficacy of multiple tumor markers and constructed a prognostic model for stage I-III TNBC patients. Patients and Methods . We included stage I-III TNBC patients whose serum tumor markers levels were measured prior to the treatment. The optimal cut-off value of each tumor marker was determined by X-tile. Then, we adopted two survival models (lasso Cox model and random survival forest model) to build the prognostic model and AUC values of the time-dependent receiver operating characteristic (ROC) were calculated. The Kaplan-Meier method was used to plot the survival curves and the log-rank test was used to test whether there was a significant difference between the predicted high-risk and low-risk groups. We used univariable and multivariable Cox analysis to identify independent prognostic factors and did subgroup analysis further for the lasso Cox model. Results . We included 258 stage I-III TNBC patients. CEA, CA125, and CA211 showed independent prognostic value for DFS when using the optimal cut-off values; their HRs and 95% CI were as follows: 1.787 (1.056–3.226), 2.684 (1.200–3.931), and 2.513 (1.567–4.877). AUC values of lasso Cox model and random survival forest model were 0.740 and 0.663 for DFS at 60 months, respectively. Both the lasso Cox model and random survival forest model demonstrated excellent prognostic value. According to tumor marker risk scores (TMRS) computed by the lasso Cox model, the high TMRS group had worse DFS (HR = 3.138, 95% CI: 1.711–5.033, p < 0.0001) and OS (3.983, 1.637–7.214, p = 0.0011) than low TMRS group. Furthermore, subgroup analysis of N 0 -N 1 patients in the lasso Cox model indicated that TMRS still had a significant prognostic effect on DFS (2.278, 1.189–4.346) and OS (2.982, 1.110–7.519). Conclusions . Our study indicated that pretreatment levels of serum CEA, CA125, and CA211 had independent prognostic significance for TNBC patients. Both lasso Cox model and random survival forest model that we constructed based on tumor markers could strongly predict the survival risk. Higher TMRS was associated with worse DFS and OS both in stage I-III and N 0 -N 1 TNBC patients.

Posttraumatic infections can occur in orthopedic trauma patients, especially in open fractures. Rapid and accurate identification of pathogens in orthopedic trauma is important for clinical diagnosis and antimicrobial treatment. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been successfully used for first-line identification of pathogens grown on culture plates. However, for direct analysis of liquid clinical specimens, pre-purification of the sample is necessary. Herein, we investigated the feasibility of coupling Fc-MBL@Fe3O4 enrichment with MALDI-TOF MS profiling in the identification of pathogens in liquid-cultured samples. This method is successfully used for the identification of pathogens in a patient with an open-leg fracture obtained at sea. Pathogens were enriched by Fc-MBL@Fe3O4 from briefly pre-cultured liquid media and identified by MALDI-TOF MS. We identified an opportunistic pathogen, Vibrio alginolyticus, which is uncommon in clinical orthopedic trauma infection but exists widely in the sea. Therefore, combining Fc-MBL@Fe3O4 enrichment and MALDI-TOF MS profiling has great potential for direct identification of microbes in clinical samples.

The Breast JournalVolume 26, Issue 9 p. 1871-1873 COMMENTARY The application of intraoperative specimen mammography for margin status assessment in breast-conserving surgery: A single-center retrospective study Chen Lin MD, Chen Lin MD orcid.org/0000-0001-6783-215X Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorKai-yue Wang MD, Kai-yue Wang MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorFang Xu MD, Fang Xu MD Department of Radiology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorYan Liu MD, Yan Liu MD Department of Radiology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorJin-fan Li MD, Jin-fan Li MD Department of Pathology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorChi Pan MD, Chi Pan MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorKai-min Hu MD, Kai-min Hu MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorWen-hong Xu MD, Wen-hong Xu MD Department of Radiotherapy, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorZhi-gang Chen MD, Zhi-gang Chen MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorYi-ding Chen MD, Yi-ding Chen MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorTao Pan MD, Corresponding Author Tao Pan MD 2311318@zju.edu.cn orcid.org/0000-0003-3058-2710 Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China Correspondence Tao Pan, Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, No. 88 Jiefang Road, Hangzhou 310009, China. Email: 2311318@zju.edu.cnSearch for more papers by this author Chen Lin MD, Chen Lin MD orcid.org/0000-0001-6783-215X Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorKai-yue Wang MD, Kai-yue Wang MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorFang Xu MD, Fang Xu MD Department of Radiology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorYan Liu MD, Yan Liu MD Department of Radiology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorJin-fan Li MD, Jin-fan Li MD Department of Pathology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorChi Pan MD, Chi Pan MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorKai-min Hu MD, Kai-min Hu MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorWen-hong Xu MD, Wen-hong Xu MD Department of Radiotherapy, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorZhi-gang Chen MD, Zhi-gang Chen MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorYi-ding Chen MD, Yi-ding Chen MD Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, ChinaSearch for more papers by this authorTao Pan MD, Corresponding Author Tao Pan MD 2311318@zju.edu.cn orcid.org/0000-0003-3058-2710 Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China Correspondence Tao Pan, Department of Breast Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, No. 88 Jiefang Road, Hangzhou 310009, China. Email: 2311318@zju.edu.cnSearch for more papers by this author First published: 24 May 2020 https://doi.org/10.1111/tbj.13835 Funding information: This work was supported by the National Natural Science Foundation of China (Grant numbers: 81602716 and 81802628). Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume26, Issue9September 2020Pages 1871-1873 RelatedInformation

Lung cancer has the highest incidence and mortality rates among the malignant tumor types worldwide. Platinum‑based chemotherapy is the main treatment for advanced non‑small‑cell lung cancer (NSCLC), and epidermal growth factor receptor‑tyrosine kinase inhibitors (EGFR‑TKIs) have greatly improved the survival of patients with EGFR‑sensitive mutations. However, there is no standard therapy for treating patients who are EGFR‑TKI resistant. Combining EGFR‑TKIs and platinum‑based chemotherapy is the most popular strategy in the clinical practice. However, the synergistic mechanism between EGFR‑TKIs and platinum remains unknown. Therefore, the aim of the present study was to determine the synergistic mechanism of gefitinib (an EGFR‑TKI) and cisplatin (a main platinum‑based drug). MTT assay, apoptosis analysis, tumorsphere formation and an orthotropic xenograft mouse model were used to examine the combination effects of gefitinib and cisplatin on NSCLC. Co‑immunoprecipitation and immunofluorescence were used to identify the underlying mechanism. It was found that gefitinib could selectively inhibit EGFR from entering the nucleus, decrease DNA‑PK activity and enhance the cytotoxicity of cisplatin on NSCLC. Collectively, the results suggested that inhibition of DNA‑dependent protein kinase by gefitinib may be due to the synergistic mechanism between gefitinib and cisplatin. Thus, the present study provides a novel insight into potential biomarkers for the selection of combination therapy of gefitinib and cisplatin.

Aims: PARK2 mutation is originally associated with the progression of Parkinson's disease. In recent years, PARK2 has been reported as a tumor suppressor gene in various cancers, including lung cancer. However, the biological functions and potential molecular mechanisms of PARK2 in non-small cell lung cancer (NSCLC) are still unclear. Methods: The level of PARK2 expression in 32 tissue samples of NSCLC and matched non-tumor lung tissues was detected by Western blot, and 64 specimens of NSCLC tissues were detected by immunohistochemistry. H1299 and H460 cell lines were used to PARK2 overexpression models, and H460 cell line was also used to PARK2 knockdown model. Using cell viability, colony formation, cell cycle, apoptosis, migration, and invasion assay, the biological functions of PARK2 were evaluated and the potential molecular mechanism of PARK2 was investigated in vitro. Meanwhile, 22 nude mice were employed for in vivo studies. Results: Western blot analysis revealed a decrease of PARK2 protein expression in human NSCLC samples. Immunohistochemistry also identified a vastly reduced expression of PARK2 in NSCLC (72%) and low PARK2 expression was significantly associated with tumor histological grade, lymph node metastasis and advanced TNM stage. Overexpression of PARK2 suppressed cell proliferation, colony formation, migration, and invasion, arrested cell cycle progression in the G1 phase, and induced apoptosis in human non-small cell lines H1299 and H460 in vitro. Meanwhile, knockdown of PARK2 had the opposite biological functions. In addition, PARK2 significantly decreased the tumor volumes in subcutaneous xenograft model and reduced the incidence of metastatic tumors in the transfer model. Exploration of the molecular mechanism of PARK2 in NSCLC showed that PARK2 negatively regulated the EGFR/AKT/mTOR signaling pathway. Conclusions: PARK2 was an important tumor suppressor in NSCLC, which might inhibit cancer growth and metastases through the down regulation of the EGFR/AKT/mTOR signaling pathway.