Prognosis Of Non-small Cell Lung Cancer Patients Research Articles

MicroRNA-133a inhibits the proliferation of non-small cell lung cancer by targeting YES1.

Previous studies have reported that microRNA-133a (miR-133a) is involved in the pathogenesis of human cancers. This study investigated the effect of miR-133a on cell proliferation in non-small cell lung cancer (NSCLC). The expression of miR-133a and YES proto-oncogene 1 (YES1) was detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay. The CCK-8 assay was used to measure cell proliferation. The relationship between miR-133a and YES1 was confirmed by dual luciferase assay. Downregulation of miR-133a was identified in NSCLC and correlated with poor prognosis in NSCLC patients. Moreover, the overexpression of miR-133a inhibited proliferation of NSCLC cells. YES1 was also confirmed as a direct target of miR-133a. Downregulation of YES1 was found to inhibit cell proliferation in NSCLC. By contrast, the upregulation of YES1 abolished the inhibitory effect of miR-133a on cell proliferation in NSCLC. miR-133a inhibited cell proliferation in NSCLC by targeting YES1, indicating that miR-133a can be used as an indicator of prognosis in NSCLC patients.

328P - A retrospective analysis of patients with non-small cell lung cancer who developed drug-induced lung disorder by immune checkpoint inhibitors

BackgroundThe presence of the immune-related adverse events (irAEs) has been reported as a preferred marker of the efficacy of immune checkpoint inhibitors (ICIs) in non-small cell lung cancer (NSCLC) patients. However, there are only a few studies that focused on the relationship between the efficacy of ICIs and the severity or a high-resolution computed tomography (HRCT) scan pattern of drug-induced lung disorder (DILD). MethodsWe conducted a retrospective study of patients with NSCLC who were treated with ICIs between January 2016 and June 2019 in our institution. We especially focused on the patients who developed DILD during ICIs treatment, and analyzed the clinical information and HRCT pattern of DILD to investigate the prognosis of patients who developed DILD. Assessment of treatment response was evaluated using the response evaluation criteria in solid tumors version 1.1. Severity of DILD was classified by grade according to pneumonitis in Common Terminology Criteria for Adverse Events (CTCAE) version 5.0. ResultsTotal 182 patients were enrolled, including 20 patients who received chemotherapy with ICIs. Among the 182 patients, 32 patients (17.6 %) developed DILD and CTCAE grade 1/2/3/4/5 were 10/7/9/2/4 patients, respectively. The median progression-free survival was 100 and 93 days, (P = 0.324, Log-rank Test) and overall survival was 415 and 501 days (P = 0.0951, Log-rank Test) in patients with or without DILD, respectively. The median period from the first ICI treatment to onset of DILD was significantly shorter in the Grade 3/4/5 group than Grade 1/2 group; 22 and 72.5 days, respectively (P = 0.029, Mann-Whitney U test). Analyzing HRCT findings according to ATS/ERS/JRS/ALAT GUIDLINE, patients who presented consolidation had better prognosis than those with comprehensive GGO. In 4 patients with Grade 5 pneumonitis developed with comprehensive GGO and the period from onset of DILD to death was extremely short; median 2 days. ConclusionsOur retrospective study suggested that development of DILD was not preferred in terms of prognosis in NSCLC patients. Physician should be alerted for the patients who developed early-onset, severe DILD with comprehensive GGO during ICI treatment. Legal entity responsible for the studyHiroyuki Yamaguchi. FundingHas not received any funding. DisclosureAll authors have declared no conflicts of interest.

Jumonji domain containing 5 is a potential prognostic indicator in non-small cell lung cancer patients who received platinum-based chemotherapy.

BackgroundPlatinum-based chemotherapy is the cornerstone of non-small cell lung cancer (NSCLC) therapy. However, the molecular mechanisms and predictive markers of platinum chemoresistance have not been fully understood. Our recent study revealed that Jumonji domain containing 5 (JMJD5) expression in cells was elevated under DNA damage by alkylating agent or UV radiation, which suggests a potential role of JMJD5 in DNA damage related chemoresistance. However, the role of JMJD5 in NSCLC chemotherapy has not been reported. In this study, we demonstrated JMJD5 as a potential prognostic indicator in NSCLC patients who received platinum-based chemotherapy.MethodsJMJD5 protein expression level in tumor and adjacent normal tissues were detected by immunohistochemistry. Samples were from primary NSCLC patients who received platinum-based chemotherapy after surgical resection. Survival curves were presented by the Kaplan-Meier method and p value was acquired by log-rank test. Multivariate analysis was tested by Cox proportional-hazards regression method.ResultsElevated JMJD5 expression was found in 27.2% cases of tumor tissues (22/81), and high JMJD5 expression were significantly associated with poor overall survival time (OS) [HR =2.881 (1.774–9.121), P=0.001] and progression-free survival time (PFS) [HR =2.255 (1.417–5.886), P=0.004] in NSCLC patients who received platinum-based chemotherapy. In multivariate analyses, JMJD5 was proved to be an independent prognostic indictor for shorter OS [HR =2.339 (1.158–4.724), P=0.018] and PFS [HR =2.031 (1.095–3.767), P=0.025).ConclusionsHigh JMJD5 expression indicated a worse prognosis in NSCLC patients who received platinum-based chemotherapy, and JMJD5 may serve as a novel predictive marker in NSCLC chemotherapy.

MicroRNA-507 represses the malignant behaviors of non-small cell lung cancer via targeting zinc finger E-box binding homeobox 2.

Non-small cell lung cancer (NSCLC) is one of the most common malignancies around the world and effective therapeutic method is yet to be excavated for advance NSCLC. MicroRNA-507 (miR-507) was found to be aberrantly expressed and affected cancer cell behaviors in some types of cancers. However, the role of miR-507 in NSCLC is largely unknown. The expression, biological role, and the underlying mechanism of miR-507 in NSCLC were explored in this study. Quantitative real-time PCR (qRT-PCR) assay was applied for the detection of miR-507 in NSCLC tissues and cell lines. Cell Counting Kit-8 (CCK-8) and colony formation assays were carried out to assess the proliferative abilities of NSCLC cells. Cell invasive capabilities were determined by transwell assays. We used Dual-Luciferase reporter assays to verify the binding between miR-507 and zinc finger E-box binding homeobox 2 (ZEB2). MiR-507 was found to be downregulated in NSCLC tissues and cell lines. Low expression of miR-507 was correlated with poor prognosis of NSCLC. Overexpression of miR-507 repressed NSCLC cell invasion and proliferation. ZEB2 was predicted to be a direct downstream molecular of miR-507 and their direct binding was verified by Dual-Luciferase reporter assays. Up-regulation of ZEB2 could significantly rescue the suppressive effects of miR-507 on NSCLC cells' invasion and proliferation. MiR-507 was noticeably downregulated in NSCLC and correlated with poor prognosis of NSCLC patients. MiR-507 represses the invasion and proliferation of NSCLC via targeting ZEB2. This study indicated that miR-507 might serve as a potential therapeutic target for NSCLC.

Dissecting the m6A methylation affection on afatinib resistance in non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer deaths. Afatinib is the first-line anti-cancer agent for treatment of NSCLC. However, unexpected resistance has been a major obstacle for its clinical efficacy. In this study, we dissected afatinib resistance from the perspective of N6-Methyladenosine (m6A) modification. First, we depicted the m6A modification profiles for the afatinib resistant and sensitive NSCLC cell lines (H1299 and A549). We found that the sum enrichment scores of the resistant cell line (H1299) was much higher than that of the sensitive cell line (A549). Next, we identified the functionally m6A-modified genes, which were the intersection of the differentially m6A methylated genes and the differentially expressed genes between H1299 and A549, as well as negative correlation between m6A modification levels and gene expression levels. In addition, functional enrichment analysis of the functionally m6A-modified genes indicated that m6A methylation might modify cell cycle to affect afatinib response. Furthermore, the functionally m6A-modified genes were over-represented in the putative drug resistance-associated genes and the FDA-approved drug targets, and had significantly higher average degree and clustering coefficient than other genes in protein-protein interaction (PPI) network. We also identified five network modules, which were all related to drug resistance functions. Finally, survival analysis demonstrated that m6A modification could affect prognosis of NSCLC patients. In conclusion, we conducted a first attempt to dissect m6A methylation affection on afatinib resistance in NSCLC, and brought inspiration for the study of epigenetic roles in drug resistance.

TOB1‑AS1 suppresses non‑small cell lung cancer cell migration and invasion through a ceRNA network

Non-small cell lung cancer (NSCLC) is the leading cause of lung cancer-associated mortality. Recent studies revealed that long non-coding (lnc)RNAs have crucial roles in human cancers. The present study was the first, to the best of our knowledge, to indicate that the lncRNA transducer of ERBB2, 1-antisense 1 (TOB1-AS1) acts as a tumor suppressor in NSCLC. Knockdown of TOB1-AS1 significantly induced NSCLC cell migration, invasion and proliferation. It was also demonstrated that the higher expression of TOB1-AS1 in NSCLC samples was associated with longer overall survival time. Furthermore, a TOB1-AS1-mediated competing endogenous RNA network in NSCLC was constructed, including Homo sapiens (hsa)-microRNA (miR)-27a-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27b-3p, hsa-miR-23c, dynein cytoplasmic 2 light intermediate chain 1, E4F transcription factor 1, TSPY-like 4, component of oligomeric Golgi complex 7, inositol hexakisphosphate kinase 2 and deltex E3 ubiquitin ligase 3. Of note, dysregulation of targets of TOB1-AS1 was associated with the prognosis of NSCLC patients. The present study suggested that TOB1-AS1 may serve as a novel biomarker for NSCLC.

Elevated expression of mcl-1 inhibits apoptosis and predicts poor prognosis in patients with surgically resected non-small cell lung cancer

BackgroundMcl-1, an anti-apoptotic member of bcl-2 family, together with cleaved poly (ADC-ribose) polymerase (c-PARP) can serve as a marker of cell apoptosis. Previously we reported that treatment of Mnk inhibitor CGP57380 resulted in decreased Mcl-1 expression while increased c-PARP expression in non-small cell lung cancer (NSCLC) cells. In this study, we aimed to investigate association between Mcl-1 expression and clinicopathological features of NSCLC, and their correlation between Mcl-1 and both proliferation index (PI) and apoptotic index (AI) in NSCLC patients.MethodsTissue microarrays (TMA) including 350 cases of surgically resected NSCLC were utilize and stained with Mcl-1, Ki-67 and c-PARP antibodies, PI and AI were then evaluated, respectively.ResultsHigher Mcl-1 expression and PI were observed in NSCLC compared with non-cancerous lung tissues (non-CLT), while AI was significantly lower in lung adenocarcinoma (ADC) compared with non-CLT. Additionally, Mcl-1 expression in lung ADC was evidently higher than that of in lung squamous cell carcinoma (SCC). The elevated Mcl-1 expression was associated with PI, and inversely related to AI in NSCLC. NSCLC patients with elevated Mcl-1 expression and high PI, or with high Mcl-1 expression and low AI had remarkably shorter overall survival time than these patients with low Mcl-1 expression.ConclusionsElevated expression of Mcl-1 might be inversely proportional to disease progression of NSCLC patients by promoting cell proliferation and inhibiting apoptosis, and Mcl-1 might serve as novel biomarker of poor prognosis for NSCLC patients.

Long non-coding RNA LINC00243 promotes proliferation and glycolysis in non-small cell lung cancer cells by positively regulating PDK4 through sponging miR-507.

Non-small cell lung cancer (NSCLC) is the main subtype of lung cancer. The overall survival of NSCLC patients is relatively low even after various treatments. Accumulating evidence demonstrated that long non-coding RNA (LncRNA) plays crucial roles in different biological process. However, the role of a novel LncRNA, LINC00243, in NSCLC remains unclear. We aimed to explore the biological role of LINC00243 in NSCLC. The mRNA and protein levels were determined by real-time PCR and western blot, respectively. Cell viability in vitro was detected by Cell Counting Kit-8 (CCK-8) assay and colony-formation assay. Reporter assay was used to detect the interactions between molecules, and the interaction was assessed by RNA pull-down assay. LINC00243 expression increased in human NSCLC tissues and associated with poor prognosis of NSCLC patients. LINC00243 knockdown inhibited proliferation and glycolysis of NSCLC cells. Mechanically, LINC00243 acted as a molecular sponge for miR-507, and miR-507 reversed the effect of LINC00243 on promoting proliferation and glycolysis of NSCLC cells. Moreover, LINC00243 modulated expression of endogenous miR-507-targeted PDK4. LINC00243 promotes proliferation and glycolysis in NSCLC cells by positively regulating PDK4 through sponging miR-507. LINC00243 could be the potential target for NSCLC treatment clinically.

The Effect of Diabetes Mellitus on Prognosis of Patients with Non-Small-Cell Lung Cancer: A Systematic Review and Meta-Analysis.

Purpose: To quantitatively evaluate the effect of preexisting diabetes mellitus (DM) on the outcomes of patients with non-small-cell lung cancer (NSCLC).Materials and Methods: Observational studies comparing the prognosis of NSCLC patients with and without diabetes were identified from PubMed, EMBASE, and The Cochrane Central Register of Controlled Trials (CENTRAL). We searched for studies that published in English from inception to March 30, 2019, using search terms related to diabetes and NSCLC. Pooled hazard ratio (HR) and 95% confidence interval (CI) were calculated by a random-effect model and subgroup analyses were performed.Results: In all, 17 of 1475 identified studies were finally included in the meta-analysis. The result revealed that preexisting diabetes had a significantly negative impact on the overall survival (OS) of patients with NSCLC (HR: 1.31, 95% CI: 1.12–1.54), especially in patients undergoing surgical treatment (HR: 1.46, 95% CI: 1.02–2.09) in comparison with those receiving only non-surgical treatment (HR: 1.33, 95% CI: 0.87–2.03). In addition, preexisting diabetes was more likely to be associated with a worse prognosis among Asian NSCLC patients than Western patients. Sensitivity analysis indicated that the main result was robust, and no evidence of publication bias was found.Conclusion: Preexisting DM has a negative impact on diabetic NSCLC patients’ prognosis.

P2.16-28 Retrospective Analysis of Patients with Previous Extrathoracic Malignancies After Resection of Non-Small Cell Lung Cancer

The number of patients with previous malignancies has increased because survival time is prolonged in various malignancies. Therefore, opportunities to consider surgical indications for non-small cell lung cancer (NSCLC) patients with previous malignancies are increasing. The aim of the study is to compare the backgrounds and prognosis of NSCLC patients who had the history of previous extrathoracic malignancies with no other cancer history. Our institutional database of patients after curative resection of NSCLC from 2006 to 2013 was reviewed. Patients with synchronous or metachronous multiple lung cancer were excluded. Clinicopathological features of these patients and information about survival and recurrence were collected from the electronic medical record. Overall survival (OS) and recurrence-free probability (RFP) were estimated using the Kaplan-Meier method. In a total of 249 patients, 49 (19.7%) had previous extrathoracic malignancies. The number of previous extrathoracic malignancies was 1 in 44 patients, 2 in 3 patients and 3 in 1 patient. Sixteen patients had colorectal cancers among previous extrathoracic malignancies, 10 patients had gastric cancers and 5 had hematopoietic neoplasms. If patients with previous extrathoracic malignancies were compared to those without, the median age was 69.5 vs. 65.5 (p=0.015), male sex was 71.4% vs. 66.5% (p=0.506), average of Brinkman index was 914.2 vs. 709.7 (p=0.127) and average of serum level of carcinoembryonic antigen was 5.4 ng/mL vs. 7.5 ng/mL (p=0.506). Pathologically, stage 0 and I NSCLC account for 73.5% of patients with previous extrathoracic malignancies and 66.0% of those without it (p=0.311). The 5-year OS was 70.3% vs. 79.1% (p=0.422) and the 5-year RFP was 58.9% vs. 67.7% (p=0.223). Patients with previous extrathoracic malignancies are significantly older than those without previous extrathoracic malignancies. Patients with previous extrathoracic malignancies had relatively earlier stage NSCLC. The OS and RFP also tended to be worse in patients with previous extrathoracic malignancies. Further study is needed to consider surgical indication for NSCLC with previous extrathoracic malignancies.

P1.01-29 MLPH Promotes Epithelial–Mesenchymal Transition and Brain Metastasis via Cdc42/PAK1 Signaling in Non-Small Cell Lung Cancer

Brain metastasis (BM) is associated with poor prognosis, recurrence, and death in patients with non-small cell lung cancer (NSCLC). Therefore, a better understanding of molecular mechanisms underlying NSCLC development and progression could provide helpful insights for NSCLC prevention and effective treatment. Melanophilin (MLPH), an actin-based transport binding partner, involves in cancer progression. However, the role of MLPH in NSCLC remains unclear. Here we elucidated the functional significance, molecular mechanisms and clinical impact of MLPH in NSCLC. RNA-Sequencing was performed to identify differentially expressed genes in lung tissues of NSCLC patients with (BM+) and without BM (BM-). The expression of MLPH was examined in the serum of BM+ and BM- patients by PCR. Integrative database analysis was used to examine MLPH levels in NSCLC tissues and analyze the relationship between MLPH levels and patient survival. Lentivirus containing small hairpin (sh) RNA targeting MLPH or empty vector was designed to explore its role in NSCLC. The cell counting kit-8 assay, wound healing assay, transwell assay, flow cytometry analysis, Phalloidin staining, xenografted tumor model and brain metastasis model were used to determine the effects of MLPH on the proliferation, migration, invasion, EMT, tumorigenesis and brain metastasis of NSCLC. Western blot analysis was used to explore the underlying mechanism. High-throughput sequencing showed that MLPH mRNA was significantly differentially expressed in lung tumors between BM+ and BM- NSCLC patients. MLPH was frequently overexpressed in NSCLC tissues and cells, and high levels of MLPH correlated with poor prognosis of NSCLC patients. Silencing MLPH by shRNA suppressed NSCLC cell proliferation, migration, invasion and TGF-β-induced EMT, and triggered cell cycle arrest and apoptosis. Being in consistent with the in vitro findings, the in vivo experiment exhibited that knockdown of MLPH inhibited xenograft tumorigenesis and brain metastasis in nude mice. Mechanically, we identified TGF-β as a key downstream effector of MLPH. MLPH silencing attenuated Cdc42/PAK1 signaling activation at least in part through the downregulation of TGF-β. Furthermore, EMT phenotypes changes caused by MLPH knockdown were partially dependent on TGF-β inhibition. Our findings uncovered the role of MLPH in NSCLC progression and provided evidence for MLPH positively modulating the Cdc42/PAK1 signaling pathway to promote EMT and metastasis via TGF-β in NSCLC cells. MLPH may have the potential as a therapeutic target against metastatic NSCLC.

P2.01-45 Clinico-Molecular Characteristics and Prognostic Outcomes of TP53 Mutated Patients with Advanced Non-Small Cell Lung Cancer

TP53 mutations are the most common molecular alterations in non-small cell lung cancer (NSCLC), but prognostic role of these mutations remains elusive. The aim of this study was to analyze the clinical and molecular characteristics of TP53 mutated NSCLC patients and to evaluate a novel approach called evolutionary action (EAp53) to stratify them into high-risk or low-risk groups. 85 advanced NSCLC patients were enrolled. Variations of TP53 and other oncogenic drivers in NSCLC were identified by next-generation sequencing. EAp53 are available at http://mammoth.bcm.tmc.edu/EAp53/. Overall, 77 patients (90.6%) exhibited at least one genetic alteration. TP53 mutations were detected in 51 patients (60.0%) mainly in exons 4, 5, 6, 7, and 8. Notably, TP53 mutations were not observed in exon 4 when they coexisted with EGFR mutations. Mutant allele frequencies (MAFs) of EGFR sensitizing mutations (exon 19 deletion or L858R) were significantly higher than those of other EGFR mutations in patients harboring TP53 mutations (p = 0.012). Among TP53 mutated patients, 39.2% of them carried TP53 truncating mutations (nonsense, splice site, or frameshift), which were correlated with female gender and smokers (p = 0.026 and p = 0.024, respectively), and were potentially associated with young patients (p = 0.078), and were more common in non-DNA-binding domain of TP53 (p = 0.033). Additionally, overall survival time of 37 patients were collected, and patients with high-risk mutations stratified by EAp53 had a significantly worse OS compared to others (HR 23.15; 95% CI 4.30-124.50; p <0.001). This study is of great significance in understanding the population characteristics of TP53 mutated NSCLC patients. Furthermore, the data also indicates that evolutionary high risk TP53 mutations can identify a subset of patients with poor prognosis, suggesting that EAp53 is a novel approach and may be useful in clinical prognosis of NSCLC patients with TP53 mutations.

PC02.04 IO Should Be Given with Chemo

PC02.04 IO Should Be Given with Chemo

PRC1 promotes cell proliferation and cell cycle progression by regulating p21/p27-pRB family molecules and FAK-paxillin pathway in non-small cell lung cancer

Background: This study aimed to demonstrate the function and molecular mechanism of protein regulator of cytokinesis 1 ( PRC1 ) in the carcinogenesis of non-small cell lung cancer (NSCLC). Methods: Bioinformatics analysis was performed. Cell culture and plasmid construction were conducted for cell transfection. mRNA and protein expression, cell proliferation, migration, and cell cycle were detected. Mice models were also constructed. The relationship between PRC1 and the prognosis of NSCLC patients was analyzed. Results: PRC1 expression was higher in tumor tissues than adjacent non-tumor tissues (P PRC1 expression were more likely to have lymph node metastasis occur (P Kip1/p27 (P Conclusions: PRC1 could promote cell proliferation and cell cycle progression through FAK-paxillin pathway molecules and the regulation of the phosphorylation level of p21/p27-pRB family molecules. PRC1 might be a new and promising therapeutic target for NSCLC.

LINC01354 enhances the proliferation and invasion of lung cancer cells by regulating miR-340-5p/ATF1 signaling pathway

Recent studies showed that long non-coding RNAs (lncRNAs) could play critical roles in tumors progression. However, the performance of LINC01354 is still limited in non-small cell lung cancer (NSCLC). In the current study, our results showed that LINC01354 was significantly increased in NSCLC tissues and cell lines. High LINC01354 expression was associated with advanced TNM stage and poor prognosis in NSCLC patients. Loss-of-function assays revealed that knockdown of LINC01354 reduced lung cancer cells proliferation and invasive ability in vitro. Subsequently, mechanism studies showed that LINC01354 positively regulated the ATF1 expression via competitive binding to miR-340-5p. Therefore, our results illustrated that LINC01354 might act as an oncogenic role by modulating the miR-340-5p/ATF1 axis, providing a novel therapeutic therapy for NSCLC treatment.

High mortality and poor treatment efficacy of immune checkpoint inhibitors in patients with severe grade checkpoint inhibitor pneumonitis in non-small cell lung cancer.

BackgroundThe treatment efficacy of immune checkpoint inhibitor (ICI) and clinical outcomes in patients with non‐small cell lung cancer (NSCLC) who develop severe grade checkpoint inhibitor pneumonitis (CIP) are unclear. Here, we report on the treatment efficacy of ICI and prognosis in NSCLC patients with severe grade CIP.MethodsIn this retrospective cohort study, CIP severity, CIP‐related mortality, and ICI efficacy in 71 patients with advanced NSCLC treated with ICIs were evaluated. Data was obtained from the patients’ medical charts.ResultsAll grade and severe grade CIP were observed in 22 and 11 patients, respectively. The CIP‐related mortality rate was 22.7% (N = 5). An Eastern Cooperative Oncology Group (ECOG) Performance Status (PS) score of ≥2 and pre‐existing interstitial lung disease (ILD) were significantly associated with the development of severe grade CIP (P = 0.001 and P = 0.035, respectively). The median progression‐free survival (PFS) and overall survival (OS) were significantly shorter in patients with severe grade CIP than in those without severe grade CIP (PFS 1.0 month, 95% confidence interval [CI] 0.5–2.0 vs. 3.5 months, 95% CI 2.0–5.0 months, P = 0.003; OS 3.0 months, 95% CI 0.5–13 vs. 12.7 months, 95% CI 8.0–21.0 months, P = 0.011).ConclusionCIP is a serious complication with a poor prognosis associated with high mortality. The efficacy of ICI is significantly worse in patients with severe grade CIP than in those without severe grade CIP. Whether ICIs should be administered to patients with CIP risk factors, such as an ECOG PS score of ≥2 or pre‐existing ILD, should be carefully assessed.

CDK5 neutralizes the tumor suppressing effect of BIN1 via mediating phosphorylation of c-MYC at Ser-62 site in NSCLC

BackgroundBridging integrator 1 (BIN1) has showed outstanding tumor-suppressive potential via inhibiting c-MYC-mediated tumorigenesis. However, a frequent phosphorylation of c-MYC at Ser-62 site could block the BIN1/c-MYC interaction and limits the tumor-suppressive effect of BIN1. Cyclin-dependent kinase 5 (CDK5), a generally dysregulated protein in various carcinomas, can mediate c-MYC phosphorylation at Ser-62 site. However, whether the existence of CDK5 could block the BIN1/c-MYC interaction remains unclear.Materials and methodsThe expression of CDK5 and BIN1 in non-small cell lung cancer (NSCLC) cell lines were measured. CDK5 was knocked down and overexpressed in H460 and PC9 cells, respectively. CCK-8, wound healing and transwell were used to detect the proliferation, migration and invasion ability of NSCLC cells. Tumor-bearing nude mouse model was built with H460 cells. Dinaciclib was added to realize the effect of CDK5 inhibition in vivo. NSCLC and matched para-carcinoma specimens were collected from 153 patients who underwent radical operation. IHC was performed to determine the expression of CDK5 in the specimens. Kaplan–Meier analysis was used to analyze the correlation between the postoperative survival and CDK5 expression.ResultsCDK5 was highly expressed in H460 cells, and knockdown of CDK5 could restore the BIN1/c-MYC interaction. Meanwhile, low expression of CDK5 was observed in PC9 cells, and overexpression of CDK5 blocked the BIN1/c-MYC interaction. Consequently, the growth, migration, invasion and epithelial mesenchymal transition (EMT) ability of H460 and PC9 cells could be facilitated by CDK5. The addition of CDK5 inhibitor Dinaciclib significantly suppressed the tumorigenesis ability of NSCLC cells in tumor-bearing mouse model. Furthermore, high expression of CDK5, along with low expression of BIN1, could predict poor postoperative prognosis of NSCLC patients. The patients with high expression of CDK5 and low expression of BIN1 showed similar prognosis, indicating that CDK5 could neutralize the tumor suppressing effect of BIN1 in clinical situation.ConclusionsCDK5 blocked the interaction of BIN1 and c-MYC via promoting phosphorylation of c-MYC at ser-62 site, ultimately facilitated the progression of NSCLC.

PRC1 promotes cell proliferation and cell cycle progression by regulating p21/p27-pRB family molecules and FAK-paxillin pathway in non-small cell lung cancer.

BackgroundThis study aimed to demonstrate the function and molecular mechanism of protein regulator of cytokinesis 1 (PRC1) in the carcinogenesis of non-small cell lung cancer (NSCLC).MethodsBioinformatics analysis was performed. Cell culture and plasmid construction were conducted for cell transfection. mRNA and protein expression, cell proliferation, migration, and cell cycle were detected. Mice models were also constructed. The relationship between PRC1 and the prognosis of NSCLC patients was analyzed.ResultsPRC1 expression was higher in tumor tissues than adjacent non-tumor tissues (P<0.05). Cells transfected with the high-expression PRC1 plasmid (TOPO-PRC1 group) had the stronger ability of proliferation and migration (P<0.05) along with a lower incidence of stay at the G2/M phase (P<0.05) than the low-expression PRC1 plasmid. Mice models showed tumors obtained from mice in the TOPO-PRC1 group significantly grew faster, larger, and heavier (P<0.05) than the low-expression PRC1 group. Among the 150 NSCLC patients, patients with the higher PRC1 expression were more likely to have lymph node metastasis occur (P<0.05) and progress into an advanced stage (P<0.05), and showed shorter survival (P<0.05). Moreover, the TOPO-PRC1 group had a lower phosphorylation level, and a lower expression of Cip1/p21 (P<0.05) and Kip1/p27 (P<0.01).ConclusionsPRC1 could promote cell proliferation and cell cycle progression through FAK-paxillin pathway molecules and the regulation of the phosphorylation level of p21/p27-pRB family molecules. PRC1 might be a new and promising therapeutic target for NSCLC.

Prognostic impact of the Controlling Nutritional Status score in patients with non-small cell lung cancer treated with pembrolizumab.

Pembrolizumab, an anti-programmed cell death-1 (PD-1) monoclonal antibody, has been shown to yield a durable response and significant survival benefit in some non-small cell lung cancer (NSCLC) patients. Recent studies have shown that the Controlling Nutritional Status (CONUT) score, a novel nutritional index, can be useful for predicting the prognosis in some malignancies. However, its usefulness in predicting the clinical outcome of immune-checkpoint inhibitor (ICI) treatment in patients with NSCLC has not been clarified. The aim of this study was to investigate the clinical significance of the CONUT score in NSCLC patients treated with pembrolizumab. We conducted a retrospective analysis of the clinical data of 32 patients with advanced NSCLC who received pembrolizumab monotherapy. A cut-off CONUT score of 2 was used to categorize patients into low and high CONUT groups. We evaluated the relation between the clinicopathological factors including CONUT score and neutrophil-to-lymphocyte ratio (NLR) and the prognosis. Twenty-two patients were classified into the low CONUT score group, while 10 were classified into the high CONUT score group. In the univariate and multivariate analyses, the number of prior treatments and the CONUT score were found to independently predict progression-free survival (PFS) (P<0.05), while the CONUT score as well as NLR was an independent prognostic factor for overall survival (P<0.05). In addition, in patients who received pembrolizumab as a first-line treatment, a high CONUT score was associated with a significantly worse PFS and overall survival in comparison to a low CONUT score. The CONUT score has potential application as a predictor of the therapeutic effect and the prognosis of NSCLC patients treated with pembrolizumab. Our findings suggest that in addition to the programmed cell death ligand 1 expression level, the CONUT may also be a useful indicator for selecting NSCLC patients who may benefit from ICI treatment.

Impact of Treatment-Related Lymphopenia on Immunotherapy for Advanced Non-Small Cell Lung Cancer

The interest in combining radiation therapy (RT) with immunotherapy is increasing. We investigated the significance of lymphopenia in patients receiving immunotherapy for non-small cell lung cancer (NSCLC), and the factors associated with treatment-related lymphopenia, with particular emphasis on RT. In this retrospective single institution study, 268 patients with advanced NSCLC received immunotherapy, of whom 146 received RT. Lymphopenia was defined as an absolute lymphocyte count <1000 cells/mm.3 Patients were divided into 2 groups depending on the presence of peri-immunotherapy lymphopenia at the start of immunotherapy or during immunotherapy. At median 6.4 months of follow-up, patients with peri-immunotherapy lymphopenia (n = 146; 54.5%) showed significantly poorer progression-free survival (PFS) (median PFS: 2.2 vs 5.9 months, P < .001) and overall survival (OS) (median OS: 5.7 vs 12.1 months, P < .001). On multivariate analysis, peri-immunotherapy lymphopenia remained a significant prognostic factor for both PFS and OS. RT significantly increased peri-immunotherapy lymphopenia with an odds ratio (OR) of 1.91 (P = .025). Factors associated with the development of RT-associated lymphopenia included multiple courses (OR, 3.78; P < .001), multiple irradiated sites (OR, 4.77; P = .018), and higher dose (≥50 Gy) (OR, 3.75; P = .004). Conversely, stereotactic body RT/radiosurgery reduced the risk (OR 0.21; P = .002). Lymphopenia was indicative of poor prognosis in NSCLC patients receiving immunotherapy and was significantly associated with more intensive RT. Choosing appropriate RT regimens and techniques may be essential in reducing lymphopenia. Promising results are expected in the era of precision RT.