CXCL12/CXCR4 facilitates perineural invasion via induction of the Twist/S100A4 axis in salivary adenoid cystic carcinoma.

The activation of CXCL12/CXCR4 axis participated in the progression of multiple cancers, but potential effect in terms of perineural invasion (PNI) in SACC remained ambiguous. In this study, we identified that CXCL12 substantially expressed in nerve cells. CXCR4 strikingly expressed in tumour cells, and CXCR4 expression was closely associated with the level of EMT-associated proteins and Schwann cell hallmarks at nerve invasion frontier in SACC. Activation of CXCL12/CXCR4 axis could promote PNI and up-regulate relative genes of EMT and Schwann cell hallmarks both in vitro and in vivo, which could be inhibited by Twist silence. After overexpressing S100A4, the impaired PNI ability of SACC cells induced by Twist knockdown was significantly reversed, and pseudo foot was visualized frequently. Collectively, the results indicated that CXCL12/CXCR4 might promote PNI by provoking the tumour cell to differentiate towards Schwann-like cell through Twist/S100A4 axis in SACC.

Interplay between cancer cells and M2 macrophages is necessary for miR-550a-3-5p down-regulation-mediated HPV-positive OSCC progression.

BACKGROUND: Human papillomavirus (HPV)-positive oral squamous cell carcinoma (OSCC) is increasing worldwide with typically higher grade and stage, while better prognosis. microRNAs (miRNAs) has been shown to play a critical role in cancer, however, their role in HPV-positive OSCC progression remains unclear. METHODS: miRNA microarray was performed to identify differentially expressed miRNAs. qRT-PCR and FISH were performed to determine the relative expression of miR-550a-3-5p. CCK-8, Flow cytometry, Wound healing, Cell invasion assays and xenograft experiments were conducted to analyze the biological roles of miR-550a-3-5p. Tumor-associated macrophages (TAMs) generation, co-culturing of cancer cells with TAMs, Western blot, Dual-luciferase reporter gene assay, Immunohistochemistry and animal studies were performed to explore the mechanisms underlying the functions of miR-550a-3-5p. RESULTS: We identified 19 miRNAs differentially expressed in HPV-positive OSCC specimens and miR-550a-3-5p was down-regulated. The low expression of miR-550a-3-5p correlated with higher tumor size and nodal metastasis of HPV-positive OSCC patients. Then, we found that miR-550a-3-5p suppressed the migration, invasion and EMT of HPV-positive OSCC cells dependent on decreasing M2 macrophages polarization. Moreover, miR-550a-3-5p, down-regulated by E6 oncoprotein, inhibited M2 macrophages polarization by YAP/CCL2 signaling, which in turn abrogating EMT program in HPV-positive OSCC cells. In addition, in both xenografts and clinical HPV-positive OSCC samples, miR-550a-3-5p levels were inversely associated with YAP, CCL2 expressions and the number of M2 macrophages. CONCLUSIONS: E6/miR-550a-3-5p/YAP/CCL2 signaling induces M2 macrophages polarization to enhance EMT and progression, revealing a novel crosstalk between cancer cells and immune cells in HPV-positive OSCC microenvironment.

Myeloid derived suppressor cells contribute to the malignant progression of oral squamous cell carcinoma.

PURPOSE: The tumor-related myeloid derived suppressor cells (MDSCs), important immunosuppressive cells in tumor microenvironment, play an important role in the cancer progression. This study is aimed to investigate the crosstalk between MDSCs and oral squamous cell carcinoma (OSCC) cells and their role in the malignant progression of OSCC. METHODS: Immunochemistry (IHC) was used to investigate the expression of CD33 in 200 OSCC, 36 premalignant. CD33+ MDSCs were sorted and enriched via magnetic-activated cell sorting (MACS) from OSCC patients or health donor, and their phenotypes were identified by flow cytometry. With a co-culture system of MDSCs and OSCC, the effects of MDSCs on OSCC proliferation, apoptosis, migration invasion, epithelial-mesenchymal transition (EMT), and vasculogenic mimicry formation (VM) formation were assessed, respectively. Besides, peripheral blood mononuclear cells (PBMCs) from health donor were cultured with OSCC supernatant, the level of MDSCs and expressions of Arginase (Arg-1) and inducible nitric oxide synthase (iNOS) were measured. RESULTS: The number of MDSCs was increased in tumor tissues of OSCC patients, and was positively related to the T stage, pathological grade, lymph node metastasis and poor prognosis. Tumor-related MDSCs of the co-culture system promoted OSCC progression by contributing to cell proliferation, migration and invasion as well as inducing EMT and VM. In turn, OSCC cells had potential to induce MDSCs differentiation from PBMCs and increase the expression of Arg-1 and iNOS. CONCLUSION: These indicated that the crosstalk between MDSCs and tumor cells facilitated the malignant progression of OSCC cells and the immune suppressive properties of MDSCs, which may provide new insights into tumor treatment on targeting tumor-associated immunosuppressive cells.

Distinguishable Prognostic miRNA Signatures of Head and Neck Squamous Cell Cancer With or Without HPV Infection.

Since their discovery in the 1990's, microRNAs (miRNA) have opened up new vistas in the field of cancer biology and are found to have fundamental roles in tumorigenesis and progression. As head and neck squamous cell carcinoma (HNSCC) with positive human papillomavirus (HPV+) is significantly distinct from its HPV negative (HPV-) counterpart in terms of both molecular mechanisms and clinical prognosis, the current study aimed to separately develop miRNA signatures for HPV+ and HPV- HNSCC as well as to explore the potential functions. Both signatures were reliable for the prediction of prognosis in their respective groups. Then Enrichment analysis was performed to predict the potential biological functions of the signatures. Importantly, combining previous studies and our results, we speculated that HPV+ HNSCC patients with low signature score had better immunity against the tumors and enhanced the sensitivity of therapies leading to improved prognosis, while HPV- HNSCC patients with high signature score acquired resistance to therapeutic approaches as well as dysregulation of cell metabolism leading to poor prognosis. Hence, we believe that the identified signatures respectively for HPV+ and HPV- HNSCC, are of great significance in accessing patient outcomes as well as uncovering new biomarkers and therapeutic targets, which are worth further investigation through molecular biology experiments.

STAT3 Promotes Invasion and Aerobic Glycolysis of Human Oral Squamous Cell Carcinoma via Inhibiting FoxO1.

Signal transducer and activator of transcription 3 (STAT3), a previously accepted tumor-promoting protein in various malignancies, plays a key role in the process of cancer glycolysis. However, the role and potential mechanism of STAT3 in aerobic glycolysis and progression of oral squamous cell carcinoma (OSCC) has not been explored. In the present study, we demonstrated that STAT3 knockdown remarkably inhibited migration, invasion, expressions of epithelial-mesenchymal transition (EMT) markers, and aerobic glycolysis of OSCC cells by up-regulation of FoxO1. Consistently, the expression of nuclear Tyr705-phosphorylated STAT3, an active form of STAT3, was significantly elevated in OSCC tissues compared with adjacent normal tissues, and increased nuclear staining of Tyr705-phosphorylated STAT3 was associated with metastasis and shorter overall survival. Moreover, FoxO1, which was also mainly expressed in OSCC specimens, decreased in poorly-differentiated tissues compared with the relatively well-differentiated ones, and inversely correlated with the expression of nuclear Tyr705-phosphorylated STAT3 from patients with OSCC. Hence, our findings collectively characterized the contributing role of STAT3/FoxO1 in invasion and aerobic glycolysis of OSCC cells, which may lead to the worse clinical outcome.

Non-coding RNAs as Regulators of Lymphangiogenesis in Lymphatic Development, Inflammation, and Cancer Metastasis.

Non-coding RNAs (ncRNAs), which do not encode proteins, have pivotal roles in manipulating gene expression in development, physiology, and pathology. Emerging data have shown that ncRNAs can regulate lymphangiogenesis, which refers to lymphatics deriving from preexisting vessels, becomes established during embryogenesis, and has a close relationship with pathological conditions such as lymphatic developmental diseases, inflammation, and cancer. This review summarizes the molecular mechanisms of lymphangiogenesis in lymphatic development, inflammation and cancer metastasis, and discusses ncRNAs' regulatory effects on them. Therapeutic targets with regard to lymphangiogenesis are also discussed.

EZH2 promotes invasion and tumour glycolysis by regulating STAT3 and FoxO1 signalling in human OSCC cells.

The enhancer of zeste homolog 2 (EZH2), known as a member of the polycomb group (PcG) proteins, is an oncogene overexpressed in a variety of human cancers. Here, we found that EZH2 correlated with poor survival of oral squamous cell carcinoma (OSCC) patients using immunohistochemistry staining. EZH2 overexpression led to a significant induction in tumour glycolysis, Epithelial-mesenchymal transition (EMT), migration and invasion of OSCC cells. Conversely, silencing of EZH2 inhibited tumour glycolysis, EMT, migration and invasion in OSCC cells. Ectopic overexpression of EZH2 increased phosphorylation of STAT3 at pY705 and decreased FoxO1 expression, and FoxO1 expression was enhanced when inhibiting STAT3. In addition, EZH2 overexpression led to a significant decrease in FoxO1 mRNA levels in nude mice xenograft. These results indicated that regulation of EZH2 might have the potential to be targeted for OSCC treatment.

Cathepsin B defines leader cells during the collective invasion of salivary adenoid cystic carcinoma.

Cathepsin B (CTSB) has been reported to be involved in cancer metastasis by altering extracellular matrix (ECM) remodeling and facilitating invasion. However, the contribution of CTSB to collective cell invasion in salivary adenoid cystic carcinoma (SACC) and the underlying mechanisms remain unclear. The present study demonstrated that collective cell invasion is commonly observed in SACC without a complete epithelial­mesenchymal transition signature. CTSB was found to be overexpressed in the invasive front of SACC compared to the tumor center, and was associated with a poor prognosis of patients with SACC. Subsequently, a 3D spheroid invasion assay was established in order to recapitulate the collective cell invasion of SACC and the results revealed that CTSB was only expressed in leader cells. The knockdown of CTSB by siRNA inhibited the migration and invasion of SACC­83 cells and impaired the formation of leader cells. CTSB knockdown also disrupted cytoskeletal organization, altered cell morphology and inhibited ECM remodeling by downregulating matrix metalloproteinase­9, focal adhesion kinase and Rho/ROCK function. Therefore, the present study provides evidence that CTSB may define leader cells in SACC and is required for collective cell invasion as a potential key regulator of ECM remodeling.

Hypoxia promotes vasculogenic mimicry formation by vascular endothelial growth factor A mediating epithelial-mesenchymal transition in salivary adenoid cystic carcinoma.

OBJECTIVES: To investigate the role of hypoxia in vasculogenic mimicry (VM) of salivary adenoid cystic carcinoma (SACC) and the underlying mechanism involved. MATERIALS AND METHODS: Firstly, wound healing, transwell invasion, immunofluorescence and tube formation assays were performed to measure the effect of hypoxia on migration, invasion, EMT and VM of SACC cells, respectively. Then, immunofluorescence and RT-PCR were used to detect the effect of hypoxia on VE-cadherin and VEGFA expression. And pro-vasculogenic mimicry effect of VEGFA was investigated by confocal laser scanning microscopy and Western blot. Moreover, the levels of E-cadherin, N-cadherin, Vimentin, CD44 and ALDH1 were determined by Western blot and immunofluorescence in SACC cells treated by exogenous VEGFA or bevacizumab. Finally, CD31/ PAS staining was performed to observe VM and immunohistochemistry was used to determine the levels of VEGFA and HIF-1α in 95 SACC patients. The relationships between VM and clinicopathological variables, VEGFA or HIF-1α level were analysed. RESULTS: Hypoxia promoted cell migration, invasion, EMT and VM formation, and enhanced VE-cadherin and VEGFA expression in SACC cells. Further, exogenous VEGFA markedly increased the levels of N-cadherin, Vimentin, CD44 and ALDH1, and inhibited the expression of E-cadherin, while the VEGFA inhibitor reversed these changes. In addition, VM channels existed in 25 of 95 SACC samples, and there was a strong positive correlation between VM and clinic stage, distant metastases, VEGFA and HIF-1α expression. CONCLUSIONS: VEGFA played an important role in hypoxia-induced VM through regulating EMT and stemness, which may eventually fuel the migration and invasion of SACC.

[A preliminary investigation on sexual arousal in blind males].

OBJECTIVE: To explore the perception of the female partner's beauty, stimulating factors for sexual arousal, and the acquisition of sexual knowledge in blind males and analyze the related influencing factors. METHODS: Using a self-designed questionnaire and with the help of the Disabled Persons' Federation and the Association for the Blind of Ganzhou, we conducted an investigation among 54 blind males at an average age of 33.5 years, who met the inclusion criteria and were enrolled according to the rosters provided by the above institutions. We convened the subjects in the appointed hospitals or other places for telephone or face-to-face question-and-answer interviews with qualified investigators trained previously. RESULTS: In the first phase, as the preliminary investigation, open-ended questions were asked of 21 of the subjects on the telephone or at a one-to-one interview. In the second phase, as the formal investigation, closed-ended questions were asked of the other 33 blind males at a face-to-face interview. The blind males perceived the beauty of their sexual partners mainly by the female's mild disposition and sweet voice (63.6% ï¼»21/33ï¼½), soft and satiny skin (51.5% ï¼»17/33ï¼½), sexy and well-shaped body (54.5% ï¼»18/33ï¼½), and intoxicating bodily scent (18.2% ï¼»6/33ï¼½). The stimulating factors for sexual arousal of the subjects included hearing the partner's soft and sweet voice (39.4% ï¼»13/33ï¼½), feeling her satiny skin (54.5% ï¼»18/33ï¼½), touching her sexy and well-shaped body (66.7% ï¼»22/33ï¼½), and the pleasant mood of the males (30.3% ï¼»10/33ï¼½). CONCLUSIONS: The blind males in Ganzhou perceive the beauty of their sexual partners mainly by their senses of hearing, touch and smell, and the stimulating factors for sexual arousal chiefly include the partner's sweet voice, satiny skin and well-shaped body as well as the pleasant mood of the males.

The crosstalk between lncRNA and microRNA in cancer metastasis: orchestrating the epithelial-mesenchymal plasticity.

Noncoding RNAs (ncRNAs) have been demonstrated to closely associate with gene regulation and encompass the well-known microRNAs (miRNAs), as well as the most recently acknowledged long noncoding RNAs (lncRNAs). Current evidence indicates that lncRNAs can interact with miRNAs and these interactions play crucial roles in cancer metastasis, through regulating critical events especially the epithelial-mesenchymal transition (EMT). This review summarizes the types of lncRNA-miRNA crosstalk identified to-date and discusses their influence on the epithelial-mesenchymal plasticity and clinical metastatic implication.

Organ-sparing Surgery in Treating Patients with Liposarcoma of the Spermatic Cord: Institutional Experience and Pooled Analysis.

BACKGROUND: Liposarcoma of the spermatic cord is rare and frequently misdiagnosed. The standard therapeutic approach has been radical inguinal orchiectomy with wide local resection of surrounding soft tissues. The current trend of organ preservation in the treatment of several cancers has started to evolve. Herein we present our testis-sparing surgery experience in the treatment of spermatic cord liposarcoma and a pooled analysis on this topic. MATERIALS AND METHODS: Clinical information from patient receiving organ-sparing surgery was described. Clinical studies evaluating this issue were identified by using a predefined search strategy, e.g., Pubmed database with no restriction on date of published papers. The literature search used the following terms: epidemiology , surgery , chemotherapy , radiotherapy , testis sparing surgery, spermatic cord sarcomas/ liposarcomas. RESULTS: Patient received a complete excision of the lesion, preserving the spermatic cord and the testis. The final pathological report showed a well differentiated liposarcoma with negative surgical margins and no signs of local invasion. After 2-year of follow-up, there was no evidence of local recurrence. Since the first case reported in 1952, a total of about 200 well-documented spermatic cord liposarcoma cases have been published in English literature. Among these patients, only three instances were reported to have received an organ-sparing surgery in the treatment of spermatic cord liposarcoma. CONCLUSIONS: Radical inguinal orchiectomy and resection of the tumor with a negative microscopic margin is the recommended treatment for liposarcoma of the spermatic cord. But for small, especially well-differentiated, lesions, testis-sparing surgery might be a good option if an adequate negative surgical margin is assured.

[Diagnosis and therapy of metastatic tumors in the adrenal gland--a report of 21 cases].

BACKGROUND & OBJECTIVE: Recently, the occurrence of metastasis to the adrenal gland is increasing, while the early and differentiated diagnosis still remains difficult. Whether metastasis to the adrenal gland needs to be resected and when and how the resection should be done are controversial. This study was to explore the surgical indications of metastasis to the adrenal gland and the role of laparoscopic adrenalectomy in the treatment of this disease. METHODS: Clinical data of 21 patients with metastatic tumors in the adrenal gland, treated in Cancer Center of Sun Yat-sen University from Mar. 1997 to Mar. 2004, were analyzed retrospectively. Literature of the diagnosis and therapy was reviewed. RESULTS: The diagnosis rates of ultrasonography and spiral or thin-cut computed tomography (CT) were 70.0% (7/10) and 84.6% (11/13). Ten patients received adrenalectomy with negative resection margins, 4 of them received laparoscopic adrenalectomy. The patients survived for 1-67 months, with a median of 18 months. One patient was still alive 67 months after the adrenalectomy, and 2 was lost. Eleven patients received palliative operation or no treatment, 10 of them survived for 5-28 months with a median of 13 months, while 1 was lost during follow-up. The difference in survival rates between the 2 groups was not significant (P=0.346). CONCLUSIONS: Ultrasonography and CT are important diagnosis methods for metastatic adrenal cancer. No evidence of tumor invasion revealed by preoperative imaging studies, no adjacent lymphadenopathy and no extraladrenal metastasis are indications of adrenalectomy. Laparoscopic adrenalectomy is safe and effective for those well-selected patients.

[Diagnostic value of urodynamic parameters multianalysis in the evaluation of bladder outlet obstruction resulting from benign prostatic hyperplasia].

OBJECTIVE: To construct a function model that can be used in the diagnosis bladder outlet obstruction (BOO) resulting from benign prostatic hyperplasia, and to develop a diagram allowing the judgement of bladder outlet for patients with different detrusor contractility, especially with impaired one. METHODS: Urodynamic and clinical data of 131 men were analyzed retrospectively. By Logistic analysis, a function model was constructed. Based on the model, a diagram allowing the evaluation of bladder outlet was drawn. The cutoff point for diagnosing BOO with the function model and the curve was confirmed by ROC curve analysis. RESULTS: The function model (BOOI) was obtained by the formula 5.03 x residual fraction + 0.04 x PdetatQmax - 0.20 x Qmax - 0.91 + alpha (alpha = 0 for those with low pressure-low flow on P-FS, alpha = 1.42 for high pressure-low flow, alpha = -7.30 for high pressure-high flow). The cutoff point for BOOI diagnosing BOO was 0.36. When validated, the sensitivity, specificity, positive predictive value, and negative predictive value were 85.7%, 91.7%, 96.0% and 73.3% respectively. CONCLUSION: The BOOI, with an easy calculation mode, could predict the probability of BOO. The sensitivity and specificity of the criterion for the diagnosis of BOO were satisfactory. The curve we drew could help to differentiate the obstructed men with low pressure-low flow and thus benefit them by surgical relief of their obstruction.