Mutant TP53 promotes invasion of lung cancer cells by regulating desmoglein 3.

PURPOSE: Targeted therapies have markedly improved the prognosis of lung cancer patients; nevertheless, challenges persist, including limited beneficiary populations and the emergence of drug resistance. This study investigates the molecular mechanisms of mutant TP53 in lung cancer, aiming to contribute to novel strategies for targeted therapy. METHODS: The TCGA database was employed to delineate the mutational landscape of TP53 in lung cancer patients. Differential gene expression between TP53-mutant and wild-type patients was analyzed, followed by functional enrichment. DSG3 protein expression in lung cancer patients was assessed using IHC, and its impact on prognosis was analyzed in the TCGA database. The influence of TP53 on the downstream gene DSG3 was investigated using qPCR, ChIP-qPCR, and luciferase reporter gene assays. Protein enrichment in the DSG3 promoter region was examined through IP-MS, and the regulatory role of the HIF1-α/TP53 complex on DSG3 was explored using Co-IP, luciferase assays, and ChIP-qPCR. Molecular interactions between TP53 (R273H) and HIF1-α were detected through immunoprecipitation and molecular docking. The effects and mechanisms of DSG3 on lung cancer phenotypes were assessed through WB, transwell, and wound healing assays. RESULTS: TP53 mutations were present in 47.44% of patients, predominantly as missense mutations. DSG3 exhibited high expression in TP53-mutant lung cancer patients, and this elevated expression correlated with a poorer prognosis. TP53 interference led to a reduction in DSG3 mRNA expression, with TP53 mutant P53 enriching at the P2 site of the DSG3 promoter region, a recruitment facilitated by HIF1-α. The DBD region of TP53 (R273H) demonstrated interaction with HIF1-α. DSG3, activated through Ezrin phosphorylation, played a role in promoting invasion and metastasis. CONCLUSIONS: Mutant TP53 facilitates lung cancer cell invasion by modulating desmoglein 3.

The learning curve of a bronchus-first method in bi-port video-assisted thoracoscopic surgery for left upper lobe lung cancer.

Video-assisted thoracic surgery (VATS) has been widely used in lung cancer treatment. However, VATS left upper lobectomy (LUL) is complex due to the intricate branching pattern of the left pulmonary artery (PA). Nevertheless, VATS right upper lobectomy can be simplified through a bronchus-first and simultaneous vessel stapling technique. In this study, the learning curve was obtained while ensuring favorable oncological outcomes using bronchus-first method for VATS LUL. First, retrospective data of 148 consecutive patients who underwent VATS LUL (bronchus-first method) for non-small cell lung cancer (NSCLC) from March 2018 to October 2020 were analyzed. The learning curve was then assessed via cumulative sum (CUSUM) analysis. Moreover, data at different stages of the learning curve, including operation time, blood loss, postoperative hospital stay, lymph node harvested, thoracotomy conversion, postoperative complications, endoscopic stapler consumptions, and 3 year overall survival, were recorded. The learning curve was best modeled as the equation: y = - 7.78 + 2.05x-2.23 × 10-2x2 + 6.43 × 10-5x3, with a good-to-fit test R2 = 0.97. The surgeon entered the proficient stage (59th case-148th case) after consecutive operations of 58 cases (learning stage, 1st case-58th case). Notably, more lymph nodes were harvested in the proficient stage than in the learning stage (17.69 ± 1.47 vs. 15.53 ± 1.43, P < 0.01). Compared with the learning stage, the proficient stage was associated with shorter operation time (114.28 ± 8.56 min vs. 126.81 ± 7.30 min, P < 0.01), fewer blood loss (44.22 ± 7.75 mL vs. 57.41 ± 22.98 mL, P < 0.01), shorter postoperative hospital stay (6.02 ± 0.99 d vs. 7.22 ± 1.34 d, P < 0.01), and fewer endoscopic stapler consumptions (5.89 ± 0.64 vs. 6.53 ± 0.50, P < 0.01). However, thoracotomy conversion (4/90 vs. 5/58, P = 0.32), postoperative complications (10/90 vs. 11/58, P = 0.23) and 3 year overall survival (62.2% vs. 50.8%, log-rank test, P = 0.11) showed no significant difference between the two stages. The surgeon with former single-direction VATS lobectomy experience can master bronchus-first VATS LUL after attending to 58 cases.

METTL3 suppresses invasion of lung cancer via SH3BP5 m6A modification.

The metastasis of lung cancer poses a major clinical challenge, and m6A modification has been implicated in regulating the invasive capabilities of tumor cells. However, the mechanisms underlying m6A modification in lung cancer metastasis are not well understood. This study aims to explore the biological functions and molecular mechanisms of methyltransferase-like 3 (METTL3) in lung cancer. In this study, METTL3 were found to be downregulated in lung cancer tissues. Functionally, METTL3 inhibited the migration and invasion abilities of lung cancer cells in vitro. Furthermore, SH3 domain binding protein 5 (SH3BP5) was identified as a downstream target of METTL3. Overexpression of SH3BP5 suppressed the invasive capacity of lung cancer cells, and this regulation was m6A-dependent. Finally, we discovered that YTH N6-methyladenosine RNA binding protein F1 (YTHDF1) mediated stability is responsible for maintaining the m6A modification of SH3BP5 mRNA. Overall, our study provides insights into the critical role of METTL3-mediated m6A modification and m6A-dependent regulatory mechanisms in the progression of human lung cancer. We demonstrated that METTL3 regulates the mRNA stability of SH3BP5 in a YTHDF1-dependent manner, thereby impacting the invasive capacity of lung cancer cells.

Feasibility and safety of expandable mediastinoscopic and laparoscopic radical esophagectomy.

Background: At present, minimally invasive radical esophagectomy is the main surgical method for esophageal cancer treatment, but it has inherent limitations. We have developed a novel method of radical esophagectomy without thoracotomy to improve this situation, namely, by using EMLE. We evaluated the feasibility and safety of expandable mediastinoscopic and laparoscopic radical esophagectomy (EMLE) through a retrospective analysis. Methods: From January 2019 to June 2022, we successfully performed 106 cases of radical resection of esophageal cancer with this new surgical technique, gradually improved the surgical path, and recorded the perioperative data and postoperative complications of all patients. Results: The operation was successfully performed in all patients except for two patients who required a switch to open surgery. The mean operation time was 171.11 ± 33.29 min and the mean intraoperative blood loss was 93.53 ± 56.32 ml. The mean number of removed lymph nodes was 23.59 ± 5.42. The postoperative complications included pneumonia (3.77%), recurrent laryngeal nerve palsy (1.89%), anastomotic leak (14.15%), pleural effusion (5.66%), chylothorax (2.83%), and reoperation (4.72%). All complications were graded I-III per the Clavien-Dindo classification. No perioperative death was recorded. Conclusion: Expandable mediastinoscopic and laparoscopic radical esophagectomy is feasible for radical resection of esophageal cancer, with good therapeutic effect and safety. Because of its minimal impact on patients and convenient operation, it is a novel surgical option for patients with esophageal cancer and is expected to become a standard surgical method for radical esophagectomy in the future.

Clinical Characteristics of Malignant Pulmonary Sclerosing Pneumocytoma Based on a Study of 46 Cases Worldwide.

Objective: To analyze the clinical characteristics of patients with malignant pulmonary sclerosing pneumocytoma (PSP) with metastasis, recurrence, and growth and to improve clinicians' understanding of PSP in patients with malignant tumor characteristics. Methods: A total of 46 PSP patients with malignant tumor characteristics were identified in the literature search and compared with 38 patients with benign PSP diagnosed and treated in our hospital in the past 5 years. We explored the pathogenesis, clinical symptoms, diagnostic methods, treatment strategies and prognosis of PSP patients with malignant tumor. Results: The characteristics of young age (≤41 years old), larger tumor (≥36mm), lymph node metastasis and distribution in East Asians are indicative of PSP with malignant potential. Such patients should undergo segmental resection or lobectomy, combined with necessary lymph node dissection or biopsy. All patients with PSP should have an entire course of follow-up management, because they may have an adverse prognosis such as recurrence, growth, metastasis, and even death. Conclusion: PSP has the potential for malignancy. Anatomical lobectomy or segmental resection combined with lymph node dissection should be performed in PSP with some specific characteristics. Inappropriate diagnosis and treatment may lead to poor prognosis in PSP patients.

Nutritional assessment in esophageal fast-track surgery: comparisons of 4 objective malnutrition screening tools.

BACKGROUND: This cohort study aimed to compare the performance of the 2015 diagnostic criteria for malnutrition of the European Society of Clinical Nutrition and Metabolism (ESPEN), the Nutritional Risk Screening 2002 (NRS 2002), Malnutrition Universal Screening Tool (MUST), and Short-Form of Mini-Nutritional Assessment (MNA-SF) in detecting malnutrition risk and predicting postoperative complications and the failure of early oral feeding (EOF) programs in esophageal cancer patients. METHODS: The 4 tools were used to conduct malnutrition assessments before surgery. The patients were divided into the groups of severe malnutrition and mild/moderate malnutrition and the incidences of the endpoints were observed. Multivariable logistic regression and receiver operating characteristic (ROC) curve analyses were conducted. RESULTS: Two hundred and nineteen consecutive esophageal cancer patients were included in the study. The prevalence rates of severe malnutrition as determined by the ESPEN 2015 criteria, MUST, NRS 2002, and MNA-SF were 24.7%, 29.7%, 23.7%, and 16.0%, respectively. The moderate/severe malnutrition risk screened by the MUST had a high sensitivity (100.0%) with malnutrition identified by the ESPEN 2015 criteria. In total, 42 (19.2%) patients experienced major complications, and the incidence rate of EOF failure was 7.3%. The severe malnutrition identified by the ESPEN 2015 criteria, MUST, and NRS 2002 were comparable in predicting the incidence of postoperative pulmonary complications, anastomotic leakage, readmission to intensive care units (ICUs), and EOF failure, but the ESPEN 2015 criteria was better in predicting postoperative overall complications, major complications, and delayed hospital discharge. CONCLUSIONS: The ESPEN 2015 criteria specializes in identifying severe malnutrition and is better in predicting adverse surgical outcomes; however, the MUST and NRS 2002 are better superior in detecting early malnutrition and are also valuable in the perioperative management in esophageal surgery. It is recommended that the MUST be used as the malnutrition screening tool before the ESPEN 2015 criteria is applied.

Video-assisted mediastinoscopic and laparoscopic transhiatal esophagectomy for esophageal cancer.

BACKGROUND: Mediastinoscopy was originally applied for lymph node biopsy and mediastinal tumor resection. Improved video imaging with spreadable working channels enabled mediastinoscopy for inspection and tissue biopsy in the superior mediastinum but it is rarely used in minimally invasive esophageal cancer surgery. In this prospective trial, the practicability and security of spreadable video-assisted mediastinoscopic combined with laparoscopic transhiatal esophagectomy (VAME) with video-assisted thoracoscopic esophagectomy (VATE) were compared. METHODS: A total of 200 eligible patients with esophageal squamous cell carcinoma were randomly divided into VAME or VATE groups. Early postoperative outcomes and lymph node dissection between the two groups were compared. RESULTS: The operation time was significantly shorter (164.3 ± 47.0 min vs. 265.4 ± 47.2 min, P < 0.001), the number of dissected lymph nodes was less (15.8 ± 4.5 vs. 20.3 ± 6.5, P < 0.001), and the intraoperative blood loss was also significantly reduced (94.7 ± 56.7 mL vs. 184.4 ± 65.2 mL, P < 0.001) in the VAME compared to the VATE group, respectively. The incidence of pneumonia was lower (7% vs. 29%; P < 0.001) and the length of hospital stay was shorter in the VAME group compared to the VATE group (18.0 ± 7.6 days vs. 23.2 ± 7.2, P < 0.001, respectively). The chyle leak incidence appeared to be lower in the VAME group but statistical significance was not reached (1% vs. 4%; P = 0.369). There were no differences in the incidence of anastomotic leakages and recurrent laryngeal nerve paralysis between the groups. No 30-day mortality occurred in any of the cases. CONCLUSION: VAME appears to be a practicable and secure method for esophagectomy but needs further proof of concept. CLINICAL REGISTRATION NUMBER: Registered at Chinese Clinical Trial Registry, ChiCTR1900022797.

Up-regulation of microRNA miR-101-3p enhances sensitivity to cisplatin via regulation of small interfering RNA (siRNA) Anti-human AGT4D and autophagy in non-small-cell lung carcinoma (NSCLC).

The emergence of drug resistance hinders the treatment of malignant tumors, and autophagy plays an important role in tumor chemotherapy resistance. However, its mechanism in non-small cell lung cancer (NSCLC) has not been well-researched. We aim to investigate the role of miR-101-3p in cisplatin-resistant via regulation of autophagy-related protein 4D (ATG4D) and autophagy. Cell viability, apoptosis, fluorescence intensity of GFP-LC3 and RFP-GFP-LC3 were determined using Cell Counting Kit-8 (CCK-8) assay, flow cytometry, and Laser scanning confocal microscope analysis, respectively. The levels of LC3II/LC3I, P62 and ATG4D were detected by Western blot. The results showed that the sensitivity to cisplatin in NSCLC cells was up-regulated by miR-101-3p mimics treatment, inducing promoting cell apoptosis and inhibiting autophagy. Further mechanistic study identified that ATG4D was a direct target of miR-101-3p. Moreover, ATG4D siRNA also could reverse miR-101-3p inhibitor-induced the up-regulation of ATG4D and the ration of LC3II/LC3I, the down-regulation of p62 expression. Our findings indicated that miR-101-3p could regulate sensitivity to cisplatin of NSNCC cells by regulating autophagy mediated by ATG4D. Therefore, miR-101-3p may act as a potential therapeutic target for the treatment of NSCLC.

Long non-coding RNA TRPM2-AS sponges microRNA-138-5p to activate epidermal growth factor receptor and PI3K/AKT signaling in non-small cell lung cancer.

BACKGROUND: Long non-coding RNAs (lncRNAs) can play pivotal roles in tumor progression by acting as microRNA (miRNA) sponges. This study aimed to investigate the association of a novel lncRNA, TRPM2-AS, with the miR-138-5p/EGFR axis in the development of non-small cell lung cancer (NSCLC). METHODS: Sixty NSCLC tissues and paired adjacent non-tumor tissues were analyzed. The relative expression levels of TRPM2-AS, miR138-5p, and epidermal growth factor receptor (EGFR) and the interactions between them were analyzed. The NSCLC cell lines NCI-H1299 and A549 were transfected with TRPM2-AS shRNA/pcDNA, and miR-138-5p mimics. Cell proliferation, migration, invasion, and apoptosis were examined in response to different transfection conditions. Dual-luciferase reporter assay was performed to identify the target interactions between TRPM2-AS, miR-138-5p, and EGFR. A549 cells stably transfected with shRNA were injected into BALB/c null nude mice to establish a tumor xenograft model. RESULTS: TRPM2-AS was up-regulated in NSCLC tumors and cell lines. Cell proliferation, migration, and invasion were inhibited in NSCLC cells treated with sh-TRPM2-AS, while apoptosis was induced. The targeting of TRPM2-AS by miR138-5p and miR138-5p by EGFR were validated with dual-luciferase reporter assay. TRPM2-AS was found to be negatively correlated with miR138-5p but positively correlated with EGFR. PI3K/AKT/mTOR was activated by pcDNA-EGFR but inactivated by miR-138-5p mimics. In the tumor xenograft mouse model, sh-TRPM2-AS suppressed tumor formation, reduced the expression of EGFR and Ki67, and promoted tumor cell apoptosis. CONCLUSIONS: Our results suggested that TRPM2-AS can increase the levels of EGFR via sponging miR-138-3p; this promoted NSCLC cell proliferation, migration, and invasion in vitro, and exacerbated tumors in vivo. These findings highlight TRPM2-AS/miR-138-5p as a potential target for reducing drug resistance in patients with NSCLC.

Effectiveness and safety of bovine pericardium patch repair for cervical anastomotic leakage after oesophagectomy for cancer.

BACKGROUND: The objective of this study is to evaluate the effectiveness and safety of bovine pericardium patch (BPP) repair for cervical anastomotic leakage after esophageal squamous cancer. METHODS: Intractable cervical anastomotic leakage developed in 7 patients of esophageal squamous cell carcinoma undergoing cervical anastomosis. These patients received the BPP repair. The necrotic tissue around the cervical anastomosis was removed during the operation, and the defect was repaired with BPP according to the size of the leakage. RESULTS: The operative duration was 60-90 min (median, 75 min). There were no signs of recurrent anastomotic leakage in each patient undergoing BPP repair. Oral intake was initiated 5-8 days (median, 6 days) after the BPP repair operation without any discomfort. CONCLUSIONS: The BPP repair is a safe and effective processing scheme for patients with cervical anastomotic fistula after resection of esophageal squamous cell carcinoma. This method may be recommended for appropriate patients with intractable cervical anastomotic fistula.

Rotational Rheology of Bovine Serum Albumin Solutions: Confounding Effects of Impurities, Mechanistic Considerations and Potential Implications on Protein Formulation Development.

PURPOSE: To show and rationalize the confounding effects on the rotational/oscillatory rheology of surface active impurities in commercial protein formulations such as bovine serum albumin, BSA. METHODS: Bulk and interfacial rotational/oscillatory rheology were used to study the viscosity, complex viscosity, storage/elastic modulus, G' and loss/viscous modulus, G", as a function of time of aqueous formulations of BSA and their purified components. RESULTS: Viscosity/time profiles at steady shear for different commercial BSA products and lots showed viscosity increase, decrease and time-independent profiles at low shear rates. All lots showed shear thinning. BSA monomer and dimers/aggregates, in general, showed similar profiles. Addition of low levels of surfactant or high shear rates rendered all solutions to be Newtonian-like. Interfacial viscosity studies paralleled those on the rotational rheometer. G' > G" with viscosity increase and G' < G" with viscosity decrease over time. CONCLUSIONS: We provide a rational explanation for the time-dependent and source-dependent rheological behavior of aqueous formulations of commercially available BSA proteins based on the migration of protein and surface active impurities to the air/water interface within the rheometer plates leading to the formation and breakdown of protein networks. Highly purified proteins is warranted in rheological studies of protein drug product candidates.

The expression of SALL4 is significantly associated with EGFR, but not KRAS or EML4-ALK mutations in lung cancer.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths worldwide; unfortunately, its prognosis is still very poor. Therefore, developing the target molecular is very important for lung cancer diagnosis and treatment, especially in the early stage. With this in view, spalt-like transcription factor 4 (SALL4) is considered a potential biomarker for diagnosis and prognosis in cancers, including lung cancer. METHODS: In order to better investigate the association between the expression of SALL4 and driver genes mutation, 450 histopathologically diagnosed patients with lung cancer and 11 non-cancer patients were enrolled to test the expression of SALL4 and the status of driver genes mutation. This investigation included epidermal growth factor receptor (EGFR), kirsten rat sarcoma viral oncogene homolog (KRAS), and a fusion gene of the echinoderm microtubule-associated protein-like 4 (EML4) and the anaplastic lymphoma kinase (ALK). RESULTS: The results of the study showed that females harbored more EGFR mutation in adenocarcinoma (ADC). The mutation rate of KRAS and EML4-ALK was about 5%, and the double mutations of EGFR/EML4-ALK were higher than EGFR/KRAS. In the expression analysis, the expression of SALL4 was much higher in cancer tissues than normally expected, especially in tissues that carried EGFR mutation (P<0.05), however, there were no significant differences between different mutation types. Likewise, there were no significant differences between expression of SALL4 and KRAS and EML4-ALK mutations. CONCLUSIONS: SALL4 is up regulated in lung cancer specimens and harbors EGFR mutation; this finding indicates that SALL4 expression may be relevant with EGFR, which could provide a new insight to lung cancer therapy. The mechanism needs further investigation and analysis.

Clinical significance of nucleostemin and proliferating cell nuclear antigen protein expression in non-small cell lung cancer.

PURPOSE: To investigate the expression and significance of the expression of nucleostemin (NS) and proliferating cell nuclear antigen (PCNA) protein in non-small cell lung cancer (NSCLC). METHODS: Immunohistochemistry (streptavidin-peroxidase method) was used to detect NS and PCNA expression in 53 NSCLC samples and 15 normal lung samples. RESULTS: NS protein expression was detected in 54.7% (29/53) of the NSCLC samples and 0% (0/15) of the normal lung samples (p<0.01). Furthermore, the positive expression rate of PCNA was 6.67% (1/15) in normal lung samples and 71.7% (38/53) in NSCLC samples (p<0.05). Also, the NS protein expression rate was 65.2% (15/23) in adenocarcinoma tissue samples, significantly higher than that in squamous tissues, where the NS expression rate was 46.7% (14/30) (p<0.05). In addition, the NS expression rate of 42.9% (15/35) in well or moderately differentiated tumor tissues was lower than the rate of 77.8% (14/18) in poorly differentiated tumor tissues (p<0.05). The grade of differentiation had no correlation with tumor-node-metastasis (TNM) stage and lymph node metastasis (p>0.05). Also, the positive expression rate of PCNA was significantly higher in NSCLC samples than in normal lung samples (p<0.05). In addition, a positive correlation was found between NS and PCNA expression in NSCLC (p<0.05). CONCLUSION: The highly valuable tumor molecular markers, NS and PCNA, had higher expression levels in NSCLC samples. Combined detection of NS and PCNA may be important for the early diagnosis of lung cancer and individualized therapy, having also an important role in predicting tumor prognosis.