lncRNA FLJ20021 regulates CDK1-mediated PANoptosis in a ZBP1-dependent manner to increase the sensitivity of laryngeal cancer-resistant cells to cisplatin.

In this study, we investigated the role of the newly discovered lncRNA FLJ20021 in laryngeal cancer (LC) and its resistance to cisplatin treatment. We initially observed elevated lncRNA FLJ20021 levels in cisplatin-resistant LC cells (Hep-2/R). To explore its function, we transfected lncRNA FLJ20021 and cyclin-dependent kinase 1 (CDK1) into Hep-2/R cells, assessing their impact on cisplatin sensitivity and PANoptosis. Silencing lncRNA FLJ20021 effectively reduced cisplatin resistance and induced PANoptosis in Hep-2/R cells. Mechanistically, lncRNA FLJ20021 primarily localized in the nucleus and interacted with CDK1 mRNA, thereby enhancing its transcriptional stability. CDK1, in turn, promoted panapoptosis in a ZBP1-dependent manner, which helped overcome cisplatin resistance in Hep-2/R cells. This study suggests that targeting lncRNA FLJ20021 can be a promising approach to combat cisplatin resistance in laryngeal cancer by regulating CDK1 and promoting PANoptosis via the ZBP1 pathway. These findings open up possibilities for lncRNA-based therapies in the context of laryngeal cancer.

Correlation of acoustic voice analysis and Voice Handicap Index in patients with postoperative unilateral vocal cord paralysis after thyroid surgery.

Unilateral vocal cord paralysis is frequently observed in patients who undergo thyroid surgery. This study explored the correlation between acoustic voice analysis (objective measure) and Voice Handicap Index (VHI, a self-assessment tool). One hundred and forty patients who had thyroid surgery with or without postoperative unilateral vocal cord paralysis (PVCP and NPVCP) were included. The patients were evaluated by the VHI and Dysphonia Severity Index (DSI) tools. VHI scores were significantly higher in PVCP patients than in NPVCP patients. Jitter (%) and shimmer (%) were significantly increased, whereas DSI was significantly decreased in PVCP patients. Receiver operating characteristics curve revealed that VHI scores were associated with the diagnosis of PVCP, of which VHI total score yielded an area under the curve (AUC) of 0.81. Among acoustic parameters, DSI was highly associated to PVCP (AUC=0.82, 95%CI=0.75 to 0.89). Moreover, we found a correlation between VHI scores and voice acoustic parameters. Among them, DSI had a moderate correlation with functional and VHI scores, as suggested by an R value of 0.41 and 0.49, respectively. VHI scores and acoustic parameters were associated with the diagnosis of PVCP.

Computer-assisted three-dimensional individualized extreme liver resection for hepatoblastoma in proximity to the major liver vasculature.

BACKGROUND: The management of hepatoblastoma (HB) becomes challenging when the tumor remains in close proximity to the major liver vasculature (PMV) even after a full course of neoadjuvant chemotherapy (NAC). In such cases, extreme liver resection can be considered a potential option. AIM: To explore whether computer-assisted three-dimensional individualized extreme liver resection is safe and feasible for children with HB who still have PMV after a full course of NAC. METHODS: We retrospectively collected data from children with HB who underwent surgical resection at our center from June 2013 to June 2023. We then analyzed the detailed clinical and three-dimensional characteristics of children with HB who still had PMV after a full course of NAC. RESULTS: Sixty-seven children diagnosed with HB underwent surgical resection. The age at diagnosis was 21.4 ± 18.8 months, and 40 boys and 27 girls were included. Fifty-nine (88.1%) patients had a single tumor, 39 (58.2%) of which was located in the right lobe of the liver. A total of 47 patients (70.1%) had PRE-TEXT III or IV. Thirty-nine patients (58.2%) underwent delayed resection. After a full course of NAC, 16 patients still had close PMV (within 1 cm in two patients, touching in 11 patients, compressing in four patients, and showing tumor thrombus in three patients). There were 6 patients of tumors in the middle lobe of the liver, and four of those patients exhibited liver anatomy variations. These 16 children underwent extreme liver resection after comprehensive preoperative evaluation. Intraoperative procedures were performed according to the preoperative plan, and the operations were successfully performed. Currently, the 3-year event-free survival of 67 children with HB is 88%. Among the 16 children who underwent extreme liver resection, three experienced recurrence, and one died due to multiple metastases. CONCLUSION: Extreme liver resection for HB that is still in close PMV after a full course of NAC is both safe and feasible. This approach not only reduces the necessity for liver transplantation but also results in a favorable prognosis. Individualized three-dimensional surgical planning is beneficial for accurate and complete resection of HB, particularly for assessing vascular involvement, remnant liver volume and anatomical variations.

Voice treatment of school-aged children with vocal nodules with ABCLOVE rehabilitation.

BACKGROUND: This study was to explore the effectiveness of the ABCLOVE exercise on school-aged children with vocal nodules after the treatment of budesonide. METHODS: Eighty-six school-aged children with vocal nodules were divided into control and ABCLOVE therapy groups. Subjective voice assessment and dysphonia severity index (DSI) assessment were performed before and after the 3-month of therapy. RESULTS: A significant improvement was observed in the ABCLOVE therapy group as compared with the control group (p = 0.035). ABCLOVE therapy significantly reduced the hoarseness and roughness scores in school-aged children with vocal nodules. Additionally, a significant reduction in functional score, physical score, emotional score, and total pVHI score was observed in the ABCLOVE therapy group. Moreover, acoustic parameters including jitter (%) and shimmer (%) were significantly reduced, whereas MPT and DSI were increased in school-aged children with vocal nodules who received 3 months of ABCLOVE treatment. CONCLUSION: ABCLOVE therapy displayed effectiveness on school-aged children with vocal nodules after the treatment of budesonide.

SUMO1-regulated DBC1 promotes p53-dependent stress-induced apoptosis of lens epithelial cells.

Deleted in breast cancer 1 (DBC1) was initially identified from a homozygously deleted region in human chromosome 8p21. It has been well established that DBC1 plays a dual role during cancer development. Depending on the physiological context, it can promote or inhibit tumorigenesis. Whether it plays a role in lens pathogenesis remains elusive. In the present study, we demonstrated that DBC1 is highly expressed in lens epithelial cells from different vertebrates and in retina pigment epithelial cells as well. Moreover, DBC1 is SUMOylated through SUMO1 conjugation at K591 residue in human and mouse lens epithelial cells. The SUMOylated DBC1 is localized in the nucleus and plays an essential role in promoting stress-induced apoptosis. Silence of DBC1 attenuates oxidative stress-induced apoptosis. In contrast, overexpression of DBC1 enhances oxidative stress-induced apoptosis, and this process depends on p53. Mechanistically, DBC1 interacts with p53 to regulate its phosphorylation status at multiple sites and the SUMOylation of DBC1 enhances its interaction with p53. Together, our results identify that DBC1 is an important regulator mediating stress-induced apoptosis in lens, and thus participates in control of lens cataractogenesis.

MiR-5189-3p Suppresses cell Proliferation, Invasion and Migration Through Targeting EIF5A2 in Laryngeal Squamous Cell Carcinoma.

A growing body of evidence suggests that miR-5189-3p plays a critical role in multiple diseases. This study aimed to investigate the function of miR-5189-3p in laryngeal squamous cell carcinoma (LSCC) and explore its underlying mechanisms. qRT-PCR was designed to determine the expression levels of miR-5189-3p and eukaryotic translation initiation factor 5A2 (EIF5A2), while CCK-8 assay was performed to measure the effects of miR-5189-3p on cell proliferation. Transwell assay was performed to evaluate cell invasion as well as migration, and wound healing assay was applied to demonstrate cell migratory ability. Target gene prediction and luciferase reporter assay were developed to screen the possible target gene of miR-5189-3p, and Western blot was designed to measure EIF5A2 protein expression. MiR-5189-3p was down-regulated in LSCC tissues and cell lines. Up-regulation of miR-5189-3p notably inhibited cell proliferation, invasion, and migration in HEP2 and FADU cells. EIF5A2 was the potential downstream gene of miR-5189-3p, and overexpression of miR-5189-3p apparently reduced EIF5A2 expression. Moreover, reintroduction of EIF5A2 rescued the tumor suppressive effects of miR-5189-3p. MiR-5189-3p functions as a tumor inhibitor in LSCC progression via directly regulating EIF5A2 and may be a potential therapeutic target for LSCC.

HSP90ß prevents aging-related cataract formation through regulation of the charged multivesicular body protein (CHMP4B) and p53.

Cataract is a leading ocular disease causing global blindness. The mechanism of cataractogenesis has not been well defined. Here, we demonstrate that the heat shock protein 90ß (HSP90ß) plays a fundamental role in suppressing cataractogenesis. HSP90ß is the most dominant HSP in normal lens, and its constitutive high level of expression is largely derived from regulation by Sp1 family transcription factors. More importantly, HSP90ß is significantly down-regulated in human cataract patients and in aging mouse lenses, whereas HSP90ß silencing in zebrafish causes cataractogenesis, which can only be rescued by itself but not other HSP90 genes. Mechanistically, HSP90ß can directly interact with CHMP4B, a newly-found client protein involved in control of cytokinesis. HSP90ß silencing causes upregulation of CHMP4B and another client protein, the tumor suppressor p53. CHMP4B upregulation or overexpression induces excessive division of lens epithelial cells without proper differentiation. As a result, these cells were triggered to undergo apoptosis due to activation of the p53/Bak-Bim pathway, leading to cataractogenesis and microphthalmia. Silence of both HSP90ß and CHMP4B restored normal phenotype of zebrafish eye. Together, our results reveal that HSP90ß is a critical inhibitor of cataractogenesis through negative regulation of CHMP4B and the p53-Bak/Bim pathway.

Association of the CD4+/CD8+ ratio with response to PD-1 inhibitor-based combination therapy and dermatological toxicities in patients with advanced gastric and esophageal cancer.

The host immune system affects the treatment response to immune checkpoint inhibitors and can be reflected by circulating immune cells. This study aimed to evaluate whether circulating T cell subtypes are correlated with clinical response and dermatological toxicities in patients with advanced gastric and esophageal cancer receiving PD-1 inhibitor-based combination therapy (n = 203). In the training cohort, Eastern Cooperative Oncology Group performance status (ECOG PS), PD-L1 expression, antibiotic use, and CD4+/CD8+ ratio were identified as independent prognostic factors in these patients, using a Cox regression model. A nomogram to predict the overall survival (OS) and survival probabilities was constructed using these factors. The nomogram showed good discrimination ability (C-index, 0.767) and was externally confirmed in the validation and test cohorts. Kaplan-Meier analysis showed that median OS in patients with a CD4+/CD8+ ratio ≥1.10 was 6.2 months, which was significantly shorter than that in patients with a CD4+/CD8+ ratio <1.10 (P < 0.001). Patients with a CD4+/CD8+ ratio <1.10 had a superior objective response (43.8% vs. 23.1%) and disease control (72.9% vs. 59.0%) rate, relative to those with ratio ≥ 1.10. In addition, PD-L1 expression, corticosteroid use, and CD4+/CD8+ ratio can independently predict dermatological toxicities. In conclusion, baseline CD4+/CD8+ ratio is a potential prognostic factor for patients with advanced gastric and esophageal cancer treated with PD-1 inhibitor-based combination therapy, and can independently predict dermatological toxicities. In addition, a nomogram incorporating CD4+/CD8+ ratio, ECOG PS, PD-L1 expression, and antibiotic use can predict OS with considerable accuracy.

Oxaliplatin Induces Immunogenic Cell Death in Human and Murine Laryngeal Cancer.

Background: Cisplatin resistance is observed in patients with laryngeal cancer. The present study was designed to explore the efficacy of oxaliplatin on laryngeal cancer and elucidate the underlying mechanisms. Methods: Cell viability was determined by using MTT assays. Cell apoptosis was determined by using annexin V and propidium iodide (PI) staining. Flow cytometry and immunofluorescence were applied to determine the levels of calreticulin (CALR) and DiD (1,1-dioctadecyl-3,3,3,3-tetramethylindodicarbocyanine). Flow cytometry was applied to analyze the levels of CD83, CD86, IFN-γ-producing CD8+ T cells, and CD4+CD25+FoxP3+ Tregs. The levels of adenosine triphosphate (ATP) were determined by using a chemiluminescent ATP kit and cytokines were determined by using specific enzyme-linked immunosorbent assays (ELISAs). The levels of HMGB1 were determined by using Western blot and ELISA, respectively. The xenograft animal model was constructed to evaluate the antitumor effects of oxaliplatin. Results: Oxaliplatin inhibited cell growth, promoted cell apoptosis, and induced the levels of CALR, ATP, and high mobility group box protein 1 (HMGB1) in Hep-2 cells. Oxaliplatin-treated Hep-2 cells increased the intensity of DiD and the levels of CD83 and CD86 in dendritic cells (DCs), as well as induced the supernatant IL-6 and TNF-α. Oxaliplatin-treated primary laryngeal cancer cell-pulsed DCs increased the IFN-γ-producing CD8+ T cells and suppressed CD4+CD25+FoxP3+ Tregs. In vivo data showed that oxaliplatin suppressed tumor growth and increased the populations of CD86+CD80+ and CD8+CD45+ cells in the tumor tissues. Conclusion: Treatment with oxaliplatin inhibited laryngeal cancer cells by inducing immunogenic cell death.

Comprehensive Analysis of LINC01615 in Head and Neck Squamous Cell Carcinoma: A Hub Biomarker Identified by Machine Learning and Experimental Validation.

Background: Head and neck squamous cell carcinoma (HNSCC) is one of the most common cancers, but in clinical practice, the lack of precise biomarkers often results in an advanced diagnosis. Hence, it is crucial to explore novel biomarkers to improve the clinical outcome of HNSCC patients. Methods: We downloaded RNA-seq data consisting of 502 HNSCC tissues and 44 normal tissues from the TCGA database, and lncRNA genomic sequence information was downloaded from the GENECODE database for annotating lncRNA expression profiles. We used Cox regression analysis to screen prognostic lncRNAs, the threshold as HR >1 and p value <0.05. Subsequently, three survival outcomes (overall survival, progress-free interval, and disease-specific survival)-related lncRNAs overlapped to get the common lncRNAs. The hub biomarker was identified using LASSO and random forest models. Subsequently, we used a variety of statistical methods to validate the prognostic ability of the hub marker. In addition, Spearman correlation analysis between the hub marker expression and genomic heterogeneity was conducted, such as instability (MSI), homologous recombination deficiency (HRD), and tumor mutational burden (TMB). Finally, we used enrichment analysis, ssGSEA, and ESTIMATE algorithms to explore the changes in the underlying immune-related pathway and function. Finally, the MTT assay and transwell assay were performed to determine the effect of LINC01615 silencing on tumor cell proliferation, invasion, and migration. Results: Cox regression analysis revealed 133 lncRNAs with multiple prognostic significance. The machine learning algorithm screened out the hub lncRNA with the highest importance in the RF model: LINC01615. Clinical correlation analysis revealed that the LINC01615 increased with increasing the T stage, N stage, pathology grade, and clinical stage. LINC01615 could be used as a predictor of HNSCC prognosis validating by a variety of statistical methods. Subsequently, when clinical indicators were combined with the LINC01615 expression, the visualization model (nomogram) was more applicable to clinical practice. Finally, immune algorithms indicated that LINC01615 may be involved in the regulation of lymphocyte recruitment and immunological infiltration in HNSCC, and the LINC01615 expression represented genomic heterogeneity in pan-cancer. Functionally, silencing of LINC01615 suppresses cell proliferation, invasion, and migration in HEP-2 and TU212 cells. Conclusion: LINC01615 may play an important role in the prostromal cell enrichment and immunosuppressive state and serve as a prognostic biomarker in HNSCC.

Circular RNA ZNF609 promotes laryngeal squamous cell carcinoma progression by upregulating epidermal growth factor receptor via sponging microRNA-134-5p.

Emerging evidence has revealed that aberrantly expressed circular RNAs (circRNAs) play vital roles in tumorigenesis and progression of diverse human malignancies. CircZNF609 was found to be involved in hepatocellular carcinoma, but the role and underlying mechanism of circZNF609 in laryngeal squamous cell carcinoma (LSCC) remain unclear. This study aimed to explore the molecular mechanism of circZNF609 in LSCC. qRT-qPCR was performed to detect the expression of circZNF609 and microRNA-134-5p (miR-134-5p) in LSCC. Colony formation assay, CCK-8 assay, BrdU incorporation assay, clone formation assay, transwell invasion assay and Western blot analysis were performed to evaluate LSCC cell proliferation, as well as the expression of proliferating cell nuclear antigen (PCNA) and MMP-2. Luciferase reporter assay, target gene prediction and screening were used to validate downstream target genes of circZNF609 and miR-134-5p. EGFR expression was detected by Western blot analysis and RT-qPCR. Nude mice were used to detect tumor changes. CircZNF609 was upregulated in LSCC and associated with poor survival of LSCC patients. Knockdown of circZNF609 inhibited LSCC proliferation, invasion and the expression of PCNA and matrix matalloproteinases-2 (MMP-2). CircZNF609 can regulate miR-134-5p to upregulate epidermal growth factor receptor (EGFR). In addition, knockdown of EGFR or overexpression of miR-134-5p could reverse the tumor-promoting effects of circZNF609 in LSCC. In LSCC tissues, circZNF609 was negatively correlated with miR-134-5p and positively correlated with EGFR. CircZNF609 promotes the progression of LSCC via the miR-134-5p/EGFR axis, which might be the therapeutic target of LSCC.

MYPT1/PP1-Mediated EZH2 Dephosphorylation at S21 Promotes Epithelial-Mesenchymal Transition in Fibrosis through Control of Multiple Families of Genes.

The methyltransferase EZH2 plays an important role in regulating chromatin conformation and gene transcription. Phosphorylation of EZH2 at S21 by AKT kinase suppresses its function. However, protein phosphatases responsible for the dephosphorylation of EZH2-S21 remain elusive. Here, it is demonstrated that EZH2 is highly expressed in the ocular lens, and AKT-EZH2 axis is important in TGFß-induced epithelial-mesenchymal transition (EMT). More importantly, it is identified that MYPT1/PP1 dephosphorylates EZH2-S21 and thus modulates its functions. MYPT1 knockout accelerates EMT, but expression of the EZH2-S21A mutant suppresses EMT through control of multiple families of genes. Furthermore, the phosphorylation status and gene expression modulation of EZH2 are implicated in control of anterior subcapsular cataracts (ASC) in human and mouse eyes. Together, the results identify the specific phosphatase for EZH2-S21 and reveal EZH2 dephosphorylation control of several families of genes implicated in lens EMT and ASC pathogenesis. These results provide important novel information in EZH2 function and regulation.

The E3 Ligase PIAS1 Regulates p53 Sumoylation to Control Stress-Induced Apoptosis of Lens Epithelial Cells Through the Proapoptotic Regulator Bax.

Protein sumoylation is one of the most important post-translational modifications regulating many biological processes (Flotho A & Melchior F. 2013. Ann Rev. Biochem. 82:357-85). Our previous studies have shown that sumoylation plays a fundamental role in regulating lens differentiation (Yan et al., 2010. PNAS, 107(49):21034-9.; Gong et al., 2014. PNAS. 111(15):5574-9). Whether sumoylation is implicated in lens pathogenesis remains elusive. Here, we present evidence to show that the protein inhibitor of activated STAT-1 (PIAS1), a E3 ligase for sumoylation, is implicated in regulating stress-induced lens pathogenesis. During oxidative stress-induced cataractogenesis, expression of PIAS1 is significantly altered at both mRNA and protein levels. Upregulation and overexpression of exogenous PIAS1 significantly enhances stress-induced apoptosis. In contrast, silence of PIAS1 with CRISPR/Cas9 technology attenuates stress-induced apoptosis. Mechanistically, different from other cells, PIAS1 has little effect to activate JNK but upregulates Bax, a major proapoptotic regulator. Moreover, Bax upregulation is derived from the enhanced transcription activity of the upstream transcription factor, p53. As revealed previously in other cells by different laboratories, our data also demonstrate that PIAS1 promotes SUMO1 conjugation of p53 at K386 residue in lens epithelial cells and thus enhances p53 transcription activity to promote Bax upregulation. Silence of Bax expression largely abrogates PIAS1-mediated enhancement of stress-induced apoptosis. Thus, our results demonstrated that PIAS1 promotes oxidative stress-induced apoptosis through positive control of p53, which specifically upregulates expression of the downstream proapoptotic regulator Bax. As a result, PIAS1-promoted apoptosis induced by oxidative stress is implicated in lens pathogenesis.

Anti-CD47 antibody synergizes with cisplatin against laryngeal cancer by enhancing phagocytic ability of macrophages.

Cisplatin is mainly used in late-stage or recurrent laryngeal cancer patients. However, the effect of the chemotherapy is limited due to cisplatin resistance. Therefore, we explored the synergized role of immunosuppressive mediator with cisplatin in laryngeal cancer. Cancer cells isolated from tissues of patients with laryngeal cancer were treated with cisplatin to screen the potential immunosuppressive mediator, whose synergized effects with cisplatin were explored both in vivo and in vitro. CD47 was selected for its high expression in cisplatin-treated laryngeal cancer cells. Blocking CD47 expression using its neutralizing antibody (aCD47) synergized with cisplatin to increase macrophage phagocytosis in a co-culture system of human epithelial type 2 (Hep-2) cancer cells with tumor-associated macrophages (TAMs). Moreover, aCD47 together with cisplatin prevented tumor growth by inhibiting proliferation of cancer cells and the secretion of proinflammatory cytokines, as well as by inducing the apoptosis of cancer cells and phagocytosis of TAMs in a Hep-2-implanted mouse tumor model. aCD47 synergized with cisplatin against laryngeal cancer by enhancing the phagocytic ability of TAMs, and the combined therapy of cisplatin and aCD47 might serve as a novel therapeutic strategy against laryngeal cancer.

Factors associated with the quality of life for hospitalized patients with HPV-associated oropharyngeal squamous cell carcinoma.

PURPOSE: Oropharynx squamous cell cancer (OPSCC) is a type of head and neck squamous cell carcinoma. The raising OPSCC incidence is mainly attributed to human papillomavirus (HPV). HPV-related OPSCC has a relatively good prognosis, the concerns are focused on the improvement of quality-of-life (QOL). We aimed to figure out the factors which may affect the QOL of HPV-related OPSCC patients after treatment. METHODS: This study included patients with HPV-related OPSCC. The QOL of the patients were analyzed through the administration of European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 (EORTC QLQ-C30)-Chinese version, the European Organization for Research and Treatment of Cancer Head and Neck Cancer Module-35 (EORTC QLQ-H&N-35)-Chinese version, and Eating Assessment Tool-10 (EAT-10). Multivariable regression analysis was employed to detect the influence of predictor variables on the QOL of patients. RESULTS: A total of 294 HPV-related OPSCC patients were involved in this research. The results of EORTC QLQ-C30, EORTC QLQ-H&N-35, and EAT-10 demonstrated that the treatment decreased the QOL of HPV-related OPSCC patients. Several different factors including marital status, consumption of tobacco and alcohol, tumor sites, clinical stages, therapeutic strategies, and neck dissection were proved to have influence on QOL of HPV-related OPSCC after treatment. CONCLUSION: Based on the analyzation of the QOL at baseline and after treatment, we demonstrated several factors which influenced the QOL of HPV-associated OPSCC patients after treatment. These results can make a great contribution to the improvement of the QOL after treatment.

[Analysis of Clinical Characteristics and Therapeutic Efficacy of Patients with Adult Acute Lymphoblastic Leukemia].

OBJECTIVE: To explore the clinical characteristics and therapeutic efficacy of patient with adult acute lymphoblastic leukemia(ALL). METHODS: Seventy-seven ALL patients diagnosed in the first affiliated hospital of Zhengzhou University from 2018 to 2019 were selected. The immunotyping, fusion gene and gene mutation were detected by flow cytometry, real-time quantitative polymerase chain reaction (RT-PCR) and next generation sequencing (NGS). RESULTS: Among 77 patients with ALL, 66 were B-ALL, 9 were T-ALL. CD7 and cCD3 were the most valuable for the diagnosis of T-ALL, CD19 and cCD79a were the most valuable for the diagnosis of B-ALL, and CD58, CD123 were highly expressed in B-ALL. Three fusion genes: BCR-ABL (20.8%), MLL-AF4 (5.19%) and E2A-PBX1 (2.60%) were detected by RT-PCR and 10 mutant genes were detected by NGS (the total detection rate was 33.47%). The highest mutation rates were IL-7R (6 cases), NOTCH1 (6 cases), TP53 (5 cases) and FLT-3 (4 cases). Patients with IL-7R, NOTCH1 and TP53 mutations showed poor response to induction chemotherapy. CONCLUSION: The CD123, IL-7R, NOTCH1 and TP53 may be risk factors for prognosis, however, the increase of case number and prolonging of follow-up time are needed to further confirm.

[The expression of Nrf2 and Keap1 in laryngeal carcinoma and its clinical indication].

OBJECTIVE: To investigate the role of Keap1-Nrf2-ARE pathway in laryngeal carcinoma occurrence and development. METHOD: Thirty-five laryngeal carcinoma samples and para-carcinoma tissues were taken from the patients who accepted operation treatment in our hospital from Feb 2012 to Sep 2013. The expression levels of Nrf2 and Keap1 were detected in 35 cases of laryngeal carcinoma with SP immunohistochemical methods. The data were analyzed by the SPSS 13.0 statistical software. RESULT: The expression of Nrf2 in the 35 cases of laryngeal carcinoma was 77.14%, which in the para-carcinoma tissues was 8.57%, the difference between these two groups was statistically significant (P < 0.01). The expression of Keap1 in the 35 cases of laryngeal carcinoma was 71.43%, which in the para-carcinoma tissues was 31.43%, the difference between these two groups was statistically significant (P < 0.01). The expression of Nrf2 in I -II stage was 65.00% and in III-IV stage was 93.33%, the difference was significant (P < 0.05). The expression of Keap1 in I-II stage was 55.00% and in III-IV stage was 93.33%, the difference was significant (P < 0.05). Of the 35 cases, the positive expression rate of Nrf2 and Keap1 in laryngeal carcinoma with lymphnode metastasis were both 100.00%, compared with those without lymph nodes metastasis 68.00% and 60.00%, the difference was significant (P < 0.05). The expression of Nrf2 and Keap1 had no relationship with tumor differentiation, smoking and patient age (P > 0.05). CONCLUSION: Keap1-Nrf2-ARE pathway may play an important role in progression of laryngeal squamous cell carcinoma.

[The expression of PKM2 and HIF-1α in laryngeal squamous cell carcinoma and its clinical indication].

OBJECTIVE: The purpose of this study was to investigate the expression of PKM2 and HIF-1α in laryngeal squamous cell carcinoma and to analyze their correlation in laryngeal squamous cell carcinoma. METHOD: Total 37 laryngeal carcinoma samples and para-carcinoma tissues were taken from the patients who accepted operation treatment in the Second Hospital of HeBei Medical University from 06/2013 to 06/2014. The protein expression levels of PKM2 and HIF-1α were detected with SP immunohistochemical methods. The data were analyzed by the SPSS 13.0 statistical software. RESULT: The positive expression of PKM2 in laryngeal carcinoma tissues and adjacent tissues were 62.16% and 13.15%. The difference was statistically significant (P < 0.01). The positive expression of HIF-1α in laryngeal carcinoma tissues and adjacent tissues were 64.86% and 21.62%. The difference was statistically significant (P < 0.01). The positive expression of PKM2 and HIF-1α in well differentiated laryngeal squamous cell carcinoma were both 47.83%, while in medium and poorly differentiated laryngeal squamous cell carcinoma were 85.71% and 92.86% respectively. The difference was statistically significant (P < 0.05). The positive expression of PKM2 and HIF-1α in patients with lymph metastasis were 90.00% and 100.00% respectively, 51.85% in those without lymph metastasis, the difference was statistically significant (P < 0.05). The rate of HIF-1α positive expression in I-II stage was 53.85%, 90.91% in III-IV stage. The difference between the two groups was statistically significant (P < 0.05). The expression of PKM2 and HIF-1α had no relationship with the age and smoking (P > 0.05). The expression of PKM2 was positively related with HIF-1α in laryngeal squamous cell carcinoma (P < 0.01). CONCLUSION: The expression of PKM2 and HIF-1α are related with the proliferation, invasion and metastasis of laryngeal squamous cell carcinoma. It provides a certain theoretical basis for laryngeal cancer diagnosis and screening to measure the expression of PKM2 and HIF-1α as biological indicators.

[The impact of hypoxia improvement on laryngeal squamous cell carcinoma chemotherapy].

OBJECTIVE: To evaluate the effect of hypoxia improvement in Hep-2 cell on cisplatin-induced apoptosis. METHOD: Human laryngeal squamous cell carcinoma Hep-2 cells and HIF-1α-RNAi-Hep-2 cells were cultured in normoxic, hypoxic and reoxygenation condition. The inhibition of cisplatin on cell proliferation was evaluated by MTT assay. The influence of cisplatin on cell cycle and apoptosis were detected by flow cytometry. RESULT: The inhibition of cisplatin on cell proliferation was reduced by hypoxia. After HIF-1α gene was silenced, the inhibition of cisplatin on Hep-2 cell proliferation was increased apparently, but was still interfered partly by hypoxia. Hypoxia could induce cell apoptosis decreased and enhance chemotherapeutic resistance. After reoxygenation, cell apoptosis induced by cisplatin was increased significantly (P < 0.05). HIF-1α-RNAi-Hep-2 cells under hypoxia also showed certain resistance to apoptosis but the sensitivity to cisplatin was higher than that of Hep-2 cells. When cells were returned from hypoxic condition to normoxic condition for some time, the apoptosis induced by cisplatin was increased significantly (P < 0.05). CONCLUSION: The improvement of hypoxic microenvironment with HIF-1α gene knockout could increase the sensitivity of Hep-2 cells to chemotherapy.

Application of clinical PET imaging to human laryngeal squamous cell carcinoma xenografts.

Positron emission tomography (PET) imaging with [F-18]-fluoro-2-deoxy-D-glucose (18F-FDG) is extensively applied in clinical practice. However, in animal experiments, the application of clinical PET is difficult, due to limitations in sensitivity and spatial resolution. This study aimed to determine the potential of 18F-FDG PET with regard to the imaging of human laryngeal squamous cell carcinoma (LSCC) xenografts. Twenty-seven LSCC tumor-bearing nude mice were divided randomly into seven groups which were each handled differently; the anesthetization, fasting, warming and the time point at which scanning was initiated were varied. The size of each xenograft was measured prior to conducting the scan. Using the RAMLA 3D image reconstruction method, images were acquired. The region of interest (ROI) technology was adopted to calculate target and non-target (T/N) ratios. The results were subsequently analyzed by semiquantitative analysis. The analysis showed that there was no significant correlation between tumor size and PET image quality (r=0.381, P>0.05); however, the handling conditions of the mice had a greater influence on the tumor image quality. Fasting increased 18F-FDG uptake (T/N, 1.153±0.008) to a certain degree, although the effect was unstable. By contrast, combining warming and fasting increased 18F-FDG uptake significantly (T/N, 2.0±0.29; P<0.05). The acquisition time had no impact on the tumor image quality. The study demonstrated that the application of clinical PET scanning has potential in the study of human LSCC xenografts in nude mice, and that the quality of the image of the tumor is greatly influenced by the handling conditions of the animals.