SOX9 promotes stemness in the CAL27 cell line of tongue squamous cell carcinoma.

Tongue squamous cell carcinoma (TSCC) is a prevalent form of oral malignancy, with increasing incidence. Unfortunately, the 5-year survival rate for patients has not exceeded 50%. Studies have shown that sex-determining region Y box 9 (SOX9) correlates with malignancy and tumor stemness in a variety of tumors. To investigate the role of SOX9 in TSCC stemness, we analyzed its influence on various aspects of tumor biology, including cell proliferation, migration, invasion, sphere and clone formation, and drug resistance in TSCC. Our data suggest a close association between SOX9 expression and both the stemness phenotype and drug resistance in TSCC. Immunohistochemical experiments revealed a progressive increase of SOX9 expression in normal oral mucosa, paracancerous tissues, and tongue squamous carcinoma tissues. Furthermore, the expression of SOX9 was closely linked to the TNM stage, but not to lymph node metastasis or tumor diameter. SOX9 is a crucial gene in TSCC responsible for promoting the stemness function of cancer stem cells. Developing drugs that target SOX9 is extremely important in clinical settings.

Ly6D facilitates chemoresistance in laryngeal squamous cell carcinoma through miR-509/ß-catenin signaling pathway.

Chemotherapy resistance is a major limiting factor in the cure of patients with laryngeal squamous cell carcinoma (LSCC). Lymphocyte antigen 6 superfamily member D (Ly6D) is highly expressed in various tumors, but its role and underlying molecular mechanisms in chemoresistance of LSCC cells remains largely unclear. In this study, we reveal that overexpression of Ly6D facilitates LSCC cell chemoresistance, while Ly6D silencing abolishes this phenotype. Moreover, bioinformatics analysis, PCR array, and functional analysis confirmed that activation of the Wnt/ß-catenin pathway contributes to Ly6D-mediated chemoresistance. The genetic and pharmacological inhibition of ß-catenin compromises chemoresistance mediated by Ly6D overexpression. Mechanistically, Ly6D overexpression significantly attenuates the expression of miR-509-5p, thereby unleashing its target gene CTNNB1 to activate Wnt/ß-catenin pathway and ultimately promote chemoresistance. In contrast, Ly6D augmenting ß-catenin-mediated chemoresistance in LSCC cells were reversed by ectopic expression of miR-509-5p. Furthermore, ectopic expression of miR-509-5p markedly repressed the two other targets, MDM2 and FOXM1. Taken together, these data not only reveal the key role of Ly6D/miR-509-5p/ß-catenin in chemotherapy resistance, but also provide a new strategy for the clinical treatment of refractory LSCC.

[Functional evaluation of anterolateral thigh flap and forearm flap for repairing defects after oropharyngeal cancer].

Objective:To investigate the oropharyngeal function recovery of oropharyngeal squamous cell carcinoma repaired by anterolateral thigh flap and forearm flap. Methods:Retrospective study between September 2016 and September 2020 complete line 37 cases of oropharyngeal cancer postoperative soft tissue defect of femoral anterolateral flap or forearm flap to repair the patient data, in which 22 cases, using the forearm skin flap to repair 15 cases with femoral anterolateral flap, flap survival rate of preparation, compare the two groups, the incidence of vascular crisis, The functions of swallowing, speech, and velopharyngeal closure were evaluated. Results:21 cases of forearm flaps survived, and 1 case had vascular crisis. After surgical exploration, the contralateral forearm flaps survived transplantation. Fourteen anterolateral femoral flaps survived, 2 flaps had vascular crisis, and 1 flap survived after thrombus removal. The other one was repaired with pectoralis major myocutaneous flap for necrosis. Swallowing, speech, palatopharyngeal closure and other functions of the patients after the two kinds of flap repair had higher satisfaction. Conclusion:Forearm flap and anterolateral thigh flap are good choices for soft tissue defect after oropharyngeal cancer, Must act according to the special details choice appropriate therapeutic schedule.

[Expression and clinical significance of BTG-1 in 78 patients with oral squamous cell carcinoma].

PURPOSE: To investigate the expression and clinical significance of B cell translocation gene 1 (BTG1) in oral squamous cell carcinoma(OSCC). METHODS: Immunohistochemical staining was used to detect the expression of BTG1 protein in 78 cases of OSCC tissues, 78 adjacent tissues, 20 normal oral mucosa tissues, and 80 cervical lymph nodes. Western blot and real-time quantitative PCR were used to detect BTG1 protein and mRNA expression levels in 78 OSCC tissues and adjacent tissues. Kaplan-Meier survival analysis, Log rank test and Cox regression analysis were performed with SPSS 21.0 software package. RESULTS: The expression level of BTG1 in OSCC and cervical positive lymph nodes was significantly lower than that in normal tissues and negative lymph nodes adjacent to the cancer, and the expression of BTG1 in poorly differentiated OSCC was significantly lower than that in highly differentiated OSCC (P<0.05); Survival analysis showed the progression-free survival (PFS) and overall survival time(OS) of BTG1 low-expression group were significantly lower than those of high-expression group (P<0.05). Cox regression analysis showed that the degree of tumor differentiation, cervical lymph node metastasis, and BTG1 expression all affected patients' prognosis. CONCLUSIONS: BTG1 is lowerly expressed in OSCC, with expression related to TNM stage and differentiation of OSCC but no relation with gender, age, and tumor location, including that BTG1 may be involved in the occurrence, development and prognosis of OSCC.

LY6D as a Chemoresistance Marker Gene and Therapeutic Target for Laryngeal Squamous Cell Carcinoma.

Laryngeal squamous cell carcinoma (LSCC) is a common head and neck cancer that is unresponsive to chemotherapy; therefore, understanding the causes of chemotherapy resistance is important. The cancer stem cell (CSC) theory postulates that CSCs are the source of tumor chemoresistance. We enrich laryngeal CSCs to overcome chemoresistance of LSCC. A laryngeal cancer xenograft model was established, and a low dose of cisplatin was administered until chemoresistance arose. A next-generation xenograft model was established using surviving tumor cells, and the test was repeated four times to screen for CSCs. Cell function experiments were performed on each tumor cell generation (m1, m2, m3, and m4). The m3 line, with the highest stemness, was selected for transcriptome sequencing. LY6D was selected for clinical sample validation and functional verification. LY6D expression was detected in 107 laryngeal cancer samples, with high expression in 91 of these samples. LY6D expression was correlated with pathological T and clinical stages, and with cervical lymph node metastasis. The siLY6D group exhibited reduced adhesion and chemoresistance to cisplatin, 5-fluorouracil, and paclitaxel. LY6D is upregulated in laryngeal cancer and may serve as a biomarker for chemoresistance in CSCs. Moreover, LY6D could serve as an alternative antigenic peptide in the targeted treatment of laryngeal cancer.

[Relationship between genetic polymorphism of MTHFR C677T and nonsyndromic cleft lip with or without cleft palate in Shanxi Province of China].

OBJECTIVE: To assess the association between polymorphism of methylenetetrahydrofolate reductase (MTHFR) gene C677T locus and nonsyndromic cleft lip with or without cleft palate (NSCL/P) in Shanxi Province of China. METHODS: Blood samples from 150 patients and their parents and 150 controls and their mothers were collected. The polymorphism of MTHFR gene C677T locus were analyzed by the methods of polymerase chain reaction and restriction fragment length polymorphism technique(PCR-RFLP). Case-control analysis, transmission-disequilibrium test(TDT) and haplotype-based haplotype relative risk analysis(HHRR) were used to study the correlation between the gene mutation and NSCL/P. RESULTS: Hardy-Weinberg equilibrium test results showed that, the offspring and mother genotype in the case group and the control group was not deviated from the Hardy-Weinberg law of genetic equilibrium (P > 0.05). The distribution of genotype CC, CT and TT in offspring had significant difference between the two groups (P < 0.05). In the offspring and mother of case group and the control group, the carriers of the TT genotype compared to CC genotype, the OR are greater than 1 and 95% CI do not contain 1 (offspring OR: 2.692, 95% CI: 1.319-5.495; mother OR: 2.469, 95% CI: 1.136-5.363). The distribution of C allele and T mutation gene were significantly different in the offspring and mother between the two groups (P < 0.05). The TDT test showed: chi2 = 4.507, P < 0.05. The HHRR test showed: P < 0.05. CONCLUSION: The single nucleotide polymorphism of MTHFR C677T locus is associated with the development of NSCL/P in Shanxi Province.

[Expression and its significance of Cyclin D1 in oral squamous cell carcinoma].

OBJECTIVE: To investigate the expression and significance of Cyclin D1 in oral squamous cell ma (OSCC). METHODS: A immunohistochemistry method, Envosion, was employed to test the manifesting Cyclin D1 in pathological slices of 50 OSCC cases and 10 normal cases, and the results was treated with statistical lysis. RESULTS: In 50 OSCC cases, Cyclin D1 mainly manifested in karyon, and a little in cytoplasm. manifesting rates of Cyclin D1 in the samples was 80.0%, which was significantly higher than the manifesting of 20.0% in normal oral mucous membrane (P < 0.01). The manifestation of Cyclin D1 was correlated with rent pathological grades, clinical phases and lymph node metastasis (P < 0.05). CONCLUSION: The abnormal tation of Cyclin D1 is closely related with the occurrence and development of OSCC. Therefore, it can subsidiary index for OSCC treatment and prognosis.

[Expression and significance of angiopoietins in oral squamous cell carcinoma].

OBJECTIVE: To observe the expression of angiopoietins in oral squamous cell carcinoma and relationship between the expression of angiopoietins and pathologic classification. METHODS: The expression of Ang-1 and Ang-2 protein in samples from 42 oral squamous cell carcinoma and 16 oral normal mucosa were detected by immunohistochemical technique. RESULTS: The expression of Ang-1 and Ang-2 was found in both oral squamous cell carcinoma and normal control. It was found Ang-1 expression alike in oral squamous cell carcinoma and control (P > 0.05). Ang-2 expressed at low level in control while strongly positive in oral squamous cell caroinoma and the level of Ang-2 expression in oral squamous cell caroinoma was related to pathologic classification (P < 0.05). CONCLUSION: Ang-2 expression in oral squamous cell carcinoma is related to angiogenesis and pathological classification, which is probably involved in angiogenesis regulation, promotes the development and metastasis of oral squamous cell carcinoma.