Photothermolysis Mediated by Gold Nanorods Conjugated with Epidermal Growth Factor Receptor (EGFR) Monoclonal Antibody Induces Apoptosis via the Mitochondrial Apoptosis Pathway in Laryngeal Squamous Cell Cancer.

Gold nanorods (AuNRs) have unique optical properties and biological affinity and can be used to treat tumors when conjugated with other protein molecules. Our previous studies have shown that EGFR monoclonal antibody (EGFRmAb)-modified AuNRs exert strong antitumor activity in vitro by inducing apoptosis. In this study, we tested the effects of EGFRmAb-modified AuNRs on laryngeal squamous cell cancer (LSCC) in vitro and in vivo. The in vitro results showed that EGFRmAb-modified AuNRs inhibited NP-69, BEAS-2B and Hep-2 cell growth and induced mitochondria-dependent apoptosis. The mitochondrial membrane potential was reduced, leading to the release of cytochrome C (Cyt C) and consequent activation of the intrinsic mitochondrial apoptosis pathway. Moreover, we observed that the occurrence of mitochondrial apoptosis is related to the destruction of the lysosome-mitochondria axis. To verify the effects in vivo, we also established a laryngeal tumor model in nude mice by subcutaneous transplantation. In model mice treated with EGFRmAb-modified AuNRs and irradiated with an NIR laser, tumor cell apoptosis and tumor growth were inhibited. These results suggest that EGFRmAb-modified AuNRs induced apoptosis through the intrinsic mitochondrial apoptotic pathway and are a potential candidate for cancer therapy.

Optimization of endothelial growth factor receptor monoclonal antibody-gold nanorods photothermal therapy for laryngeal squamous cell carcinoma.

It reported that heat generated by near-infrared laser irradiation of gold nanorods (AuNRs) effectively inhibited tumor cells, and the conjugate of epidermal growth factor receptor monoclonal antibody (EGFRmAb) and gold nanorods could selectively binded to the surface of cancer cell membrane expressing EGFR. However, there are few research reports on EGFRmAb-AuNRs in laryngeal squamous cell carcinoma. Therefore, our study aimed to investigate the photothermal effect of EGFRmAb modified AuNRs in inducing tumor cell death in an animal model of laryngeal squamous cell carcinoma. We showed that the conjugates of EGFRmAb and AuNRs selectively entered laryngeal squamous cell carcinoma cells. We analyzed the parameters of laser irradiation by controlling the near-infrared to optimize the condition and procedure of targeted treatment in nude mice treated with EGFRmAb and AuNRs. In addition, we examined the safety of the combined therapy. Test results showed that EGFRmAb-AuNRs inhibited the growth of Hep-2 and CNE-2 cells but not normal epithelial cells, and the semi-inhibitor concentration of EGFRmAb in Hep-2 and CNE-2 cells was 4 pmol/ml and 2 pmol/ml, respectively. AuNRs injected into the tumor and irradiated by near-infrared laser effectively inhibited tumor growth in nude mice without toxic effect in mice. We further confirmed that the apoptosis and necrosis rates of tumor cells in mice were highest under 3 W/cm2 near-infrared laser irradiation and AuNRs minimum concentration of 280 µg/kg. In conclusion, we developed a new method of targeting EGFRmAb combined with AuNRs to achieve photothermal effect in the treatment of laryngeal squamous cell carcinoma.

A Novel Prognostic Prediction Model for Colorectal Cancer Based on Nine Autophagy-Related Long Noncoding RNAs.

INTRODUCTION: Colorectal cancer (CRC) is the most common gastrointestinal cancer and has a low overall survival rate. Tumor-node-metastasis staging alone is insufficient to predict patient prognosis. Autophagy and long noncoding RNAs play important roles in regulating the biological behavior of CRC. Therefore, establishing an autophagy-related lncRNA (ARlncRNA)-based bioinformatics model is important for predicting survival and facilitating clinical treatment. METHODS: CRC data were retrieved from The Cancer Genome Atlas. The database was randomly divided into train set and validation set; then, univariate and multivariate Cox regression analyses were performed to screen prognosis-related ARlncRNAs for prediction model construction. Interactive network and Sankey diagrams of ARlncRNAs and messenger RNAs were plotted. We analyzed the survival rate of high- and low-risk patients and plotted survival curves and determined whether the risk score was an independent predictor of CRC. Receiver operating characteristic curves were used to evaluate model sensitivity and specificity. Then, the expression level of lncRNA was detected by quantitative real-time polymerase chain reaction, and the location of lncRNA was observed by fluorescence in situ hybridization. Additionally, the protein expression was detected by Western blot. RESULTS: A prognostic prediction model of CRC was built based on nine ARlncRNAs (NKILA, LINC00174, AC008760.1, LINC02041, PCAT6, AC156455.1, LINC01503, LINC00957, and CD27-AS1). The 5-year overall survival rate was significantly lower in the high-risk group than in the low-risk group among train set, validation set, and all patients (all p < 0.001). The model had high sensitivity and accuracy in predicting the 1-year overall survival rate (area under the curve = 0.717). The prediction model risk score was an independent predictor of CRC. LINC00174 and NKILA were expressed in the nucleus and cytoplasm of normal colonic epithelial cell line NCM460 and colorectal cancer cell lines HT29. Additionally, LINC00174 and NKILA were overexpressed in HT29 compared with NCM460. After autophagy activation, LINCC00174 expression was significantly downregulated both in NCM460 and HT29, while NKILA expression was significantly increased. CONCLUSION: The new ARlncRNA-based model predicts CRC patient prognosis and provides new research ideas regarding potential mechanisms regulating the biological behavior of CRC. ARlncRNAs may play important roles in personalized cancer treatment.

The Yunnan national medicine Maytenus compound inhibits the proliferation of hepatocellular carcinoma (HCC) by suppressing the activation of the EGFR-PI3K-AKT signaling pathway.

Objective: To investigate the effects of Maytenus compound on the proliferation of hepatocellular carcinoma (HCC) cells in vitro and in vivo and to explore the underlying mechanism. Methods: The half maximal inhibitory concentration (IC50) values of Maytenus compound in HepG2 and BEL-7402 cells were determined by the MTS assay. HepG2 and BEL-7402 cells were treated with different concentrations of Maytenus compound. MTS assays, colony formation assays and cell cycle analyses were performed to clarify the inhibitory effect of Maytenus compound on the proliferation of HepG2 and BEL-7402 cells in vitro. After subcutaneous injection of HepG2 cells, nude mice were randomly divided into a vehicle control group and a drug intervention group, which were intragastrically administered ddH2O or Maytenus compound, respectively. The inhibitory effect of Maytenus compound on the proliferation of HepG2 cells in vivo was analyzed using subcutaneous tumor growth curves, tumor weight, the tumor growth inhibition rate and the immunohistochemical detection of BrdU-labeled cells in S phase. The organ toxicity of Maytenus compound was initially evaluated by comparing the weight difference and organ index of the two groups of nude mice. The main proteins in the EGFR-PI3K-AKT signaling pathway were detected by Western blot after Maytenus compound intervention in vivo and in vitro. Results: Maytenus compound showed favorable antiproliferation activity against HepG2 and BEL-7402 cells with IC50 values of 79.42±11.71 µg/mL and 78.48±8.87 µg/mL, respectively. MTS assays, colony formation assays and cell cycle analyses showed that Maytenus compound at different concentration gradients within the IC50 concentration range significantly suppressed the proliferation of HepG2 and BEL-7402 cells in vitro and inhibited cell cycle progression from G1 to S phase. Additionally, Maytenus compound, at an oral dose of 2.45 g/kg, dramatically inhibited, without obvious organ toxicity, the proliferation of subcutaneous tumors formed by HepG2 cells in nude mice. In addition, the tumor growth inhibition rate for Maytenus compound was 66.94%. Furthermore, Maytenus compound inhibited the proliferation of liver orthotopic transplantation tumors in nude mice. Western blot analysis showed that Maytenus compound significantly downregulated the expression of p-EGFR, p-PI3K, and p-AKT and upregulated the expression of p-FOXO3a, p27, and p21 in vivo and in vitro. Conclusion: Maytenus compound significantly inhibited the proliferation of HCC cells in vitro and in vivo. The downregulation of the EGFR-PI3K-AKT signaling pathway and subsequent inhibition of cell cycle progression from G1 to S phase is one of the possible mechanisms. Maytenus compound has a high tumor growth inhibition rate and has no obvious organ toxicity, which may make it a potential anti-HCC drug, but the results from this study need to be confirmed by further clinical trials in HCC patients.

Long non-coding (lnc)RNA profiling and the role of a key regulator lnc-PNRC2-1 in the transforming growth factor-ß1-induced epithelial-mesenchymal transition of CNE1 nasopharyngeal carcinoma cells.

OBJECTIVES: To identify key long non-coding (lnc)RNAs responsible for the epithelial-mesenchymal transition (EMT) of CNE1 nasopharyngeal carcinoma cells and to investigate possible regulatory mechanisms in EMT. METHODS: CNE1 cells were divided into transforming growth factor (TGF)-ß1-induced EMT and control groups. The mRNA and protein expression of EMT markers was determined by real-time quantitative PCR and western blotting. Differentially expressed genes (DEGs) between the two groups were identified by RNA sequencing analysis, and DEG functions were analyzed by gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses. EMT marker expression was re-evaluated by western blotting after knockdown of a selected lncRNA. RESULTS: TGF-ß1-induced EMT was characterized by decreased E-cadherin and increased vimentin, N-cadherin, and Twist expression at both mRNA and protein levels. Sixty lncRNA genes were clustered in a heatmap, and mRNA expression of 14 dysregulated lncRNAs was consistent with RNA sequencing. Knockdown of lnc-PNRC2-1 increased expression of its antisense gene MYOM3 and reduced expression of EMT markers, resembling treatment with the TGF-ß1 receptor inhibitor LY2109761. CONCLUSION: Various lncRNAs participated indirectly in the TGF-ß1-induced EMT of CNE1 cells. Lnc-PNRC2-1 may be a key regulator of this and is a potential target to alleviate CNE1 cell EMT.

AFF4 facilitates melanoma cell progression by regulating c-Jun activity.

Melanoma is characterized by high mortality and poor prognosis due to metastasis. AFF4 (AF4/FMR2 family member 4), as a scaffold protein, is a component of the super elongation complex (SEC), and is involved in the progression of tumors, e.g., leukemia, head and neck squamous cell carcinoma (HNSCC). However, few studies on AFF4 have focused on melanoma. Here, AFF4 expression levels and clinicopathological features were evaluated in melanoma tissue samples. Then, we performed cell proliferation, migration and invasion assays in A375 and A2058 cells lines in vitro to evaluate the role of AFF4 in melanoma. The effects of AFF4 knockdown in vivo were characterized via a xenograft mouse model. Finally, the correlation between c-Jun and AFF4 protein levels in melanoma was analyzed by rescue assay and immunohistochemistry (IHC). We found that AFF4 expression was upregulated in melanoma tumor tissues and that AFF4 protein expression was also closely related to the prognosis of patients with cutaneous melanoma. Moreover, AFF4 could promote the invasion and migration of melanoma cells by mediating epithelial to mesenchymal transition (EMT). AFF4 might regulate c-Jun activity to promote the invasion and migration of melanoma cells. Importantly, c-Jun was regulated by the AFF4 promoted melanoma tumorigenesis in vivo. Taken together, AFF4 may be a novel oncogene that promotes melanoma progression through regulation of c-Jun activity.

FAM136A immunoreactivity is associated with nodal involvement and survival in lung adenocarcinoma in a Chinese case series.

Lung cancer patients with lymph node metastasis usually had short overall survival and occurred distant metastases at the early stage. However, some of these people did have more prolonged survival. The underlying reason is still unclear. In this study, we found a novel molecule, family with sequence similarity 136, member A gene (FAM136A). First, we performed immunohistochemistry for FAM136A in 177 lung carcinoma tissues. Second, we carried out in vitro studies by using A549 and PC-9. We detected FAM136A immunoreactivity in 79 out of 177 (44.6%) lung carcinoma tissues, and the FAM136A status was significantly associated with tumor T stage, lymph node metastasis, and the Tumor-Node-Metastasis (TNM) staging system in these cases. Importantly, it was significantly associated with the overall survival of the patients with lymph node metastasis, especially FAM136A positive patients, who had worse outcomes. Subsequent in vitro experiments revealed that the proliferation activity and migration property decreased both A549 and PC-9 lung carcinoma cells transfected with siRNA-FAM136A, and apoptosis reduced. Meanwhile, the expression of CDK4 and CDK6 decreased. FAM136A status would be a potent, worse prognostic factor in lung cancer patients with lymph node metastasis. It would play a vital role in the proliferation, apoptosis, and migration properties of A549 and PC-9. In the future, We will focus on the uncovered signal mechanism between FAM136A and lung cancer.

CD19 and CD30 CAR T-Cell Immunotherapy for High-Risk Classical Hodgkin's Lymphoma.

BACKGROUND: In clinical applications of CAR T-cell therapy, life-threatening adverse events including cytokine release syndrome and neurotoxicity can lead to treatment failure. Outcomes of patients treated with anti-CD30 CAR T- cell have been disappointing in relapsing/refractory (r/r) classical Hodgkin's Lymphoma (cHL). METHODS: In order to understand the applicable population of multiple CAR T-cell therapy, we examined the expression of CD19, CD20, and CD30 by immunohistochemistry (IHC) in 38 paraffin-embedded specimens of cHL. In the past two years, we found only one patient with cHL who is eligible for combined anti-CD19 and CD30 CAR T-cell treatment. This patient's baseline characteristics were prone to severe adverse events. We treated this patient with low doses and multiple infusions of anti-CD19 and CD30 CAR T-cell. RESULTS: The positive expression of CD19+ + CD30+ in Reed-Sternberg (RS) cells is approximately 5.2% (2/38). The patient we treated with combined anti-CD19 and CD30 CAR T-cell did not experience severe adverse events related to CAR T-cell therapy and received long term progression-free survival (PFS). CONCLUSION: For high risk r/r cHL patients, low doses of CAR T-cell used over different days at different times might be safe and effective. More clinical trials are warranted for CD19 and CD30 CAR T-cell combination therapy.

GLT8D1 overexpression as a novel prognostic biomarker in human cutaneous melanoma.

Aberrant glycosylation plays a major role in the progression of melanoma, but little is known about glycosyltransferases. Glycosyltransferase 8 domain containing 1 (GLT8D1) is located in the Golgi apparatus and is related to transferase activity in mammals. However, its role in cancer remains unclear. The aim of this study was to investigate the expression of GLT8D1 in human melanoma and explore the relationship between GLT8D1 expression and the clinicopathological characteristics of melanoma patients via GEO data analysis combined with clinical patient data. The analysis of 45 malignant melanoma samples and 18 benign nevus samples from the GEO database was performed. Moreover, 67 patients with cutaneous melanoma and 38 patients with mucosal melanoma as well as 40 benign nevus samples were collected for our study. Immunohistochemistry analyses were implemented to evaluate GLT8D1 expression at protein level. The GEO data analysis exhibited that the GLT8D1 mRNA expression was upregulated in the melanoma samples compared with the benign nevus samples. Likewise, GLT8D1 protein expression in the cutaneous melanoma and mucosal melanoma samples was significantly higher than that in the benign nevus tissue samples (P = 0.001 and 0.046, respectively). Furthermore, the GLT8D1 protein expression in cutaneous melanoma was higher than that in mucosal melanoma (P = 0.001). The high GLT8D1 protein expression was remarkably correlated with Clark level (P = 0.027), AJCC stage (P = 0.003), ulceration status (P = 0.041), Ki-67 expression (P = 0.030) and especially with histopathological type (P = 0.001). The results of the Kaplan-Meier survival and Cox regression analyses revealed that cutaneous melanoma patients with high GLT8D1 expression (P = 0.036), Clark level (P = 0.018) and advanced AJCC stage (P = 0.003) encountered poor overall survival. Overall survival (P = 0.040) and progression-free survival (P = 0.019) were worse for the patients with high GLT8D1 expression than for the patients with low expression. These data implied that GLT8D1 could be an independent prognostic factor for an unfavorable prognosis in cutaneous malignant melanoma patients and that GLT8D1 overexpression might serve as a novel prognostic biomarker.

WD repeat and SOCS box containing protein 2 in the proliferation, cycle progression, and migration of melanoma cells.

BACKGROUND: WD repeat and SOCS box containing protein (WSB) molecules have important roles in tumorigenesis. WSB1 is dysfunctional in many malignancies. However, the effects of WSB2 in tumors, including melanoma, have not been reported. Here, we investigated the effects of WSB2 in melanoma cell proliferation, cycle progression, and migration, and the underlying mechanisms. METHODS: First, WSB2 expression levels and their association with clinicopathological features were evaluated in human melanoma tissue samples. Then, WSB2 was knocked down, using specific shRNA, in melanoma A375 and G361 cells. Proliferation, cycle progression, and migration of A375 and G361 cells were evaluated by 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-etrazolium, inner salt (MTS), colony formation, 5-ethynyl-2'-deoxyuridine (EdU), cycle, and transwell assays. The effects of WSB2 knockdown on melanoma in vivo were determined using a xenograft mouse model. To investigate the underlying mechanisms, levels of c-Myc, ß-catenin, phosphorylated retinoblastoma (p-Rb), cyclin-dependent kinase 4 (CDK4), and Cyclin D3 proteins were determined by western blotting in melanoma A375 cells with WSB2 knocked down. Furthermore, ß-catenin agonism, SKL2001, was used to evaluate the mechanisms by which knockdown of WSB2 regulated cell proliferation. RESULTS: WSB2 levels were high and they were associated with clinicopathological features in patients with melanoma. shRNA-mediated knockdown of WSB2 could inhibit proliferation, both in vivo and in vitro. Cycle progression and migration of A375 and G361 cells were also significantly inhibited by WSB2 knockdown. Moreover, down-regulation of WSB2 decreased the levels of c-Myc, ß-catenin, p-Rb, Cyclin-dependent kinase 4 (CDK4), and Cyclin D3 in melanoma G361 cells with WSB2 knocked down. Moreover, SKL2001 could effectively rescue WSB2 knockdown-mediated inhibition of cell proliferation in melanoma. CONCLUSION: This is the first report to demonstrate the effects of WSB2 on melanoma cell function. WSB2 has potential to become a new therapeutic target in patients with melanoma.

Anticancer effect of the traditional Chinese medicine herb Maytenus compound via the EGFR/PI3K/AKT/GSK3ß pathway.

BACKGROUND: Cancer is a leading cause of death worldwide; folk anticancer medicinal plants have applied for cancer treatment. The Maytenus compound tablet as traditional Chinese compound medicine has been approved for alleviating hyperplasia of mammary glands, whether it can inhibit cancer still unknown. The study was to evaluate the anticancer activity of the Maytenus compound tablet. METHODS: MTS assay evaluated the anti-proliferation effect of the Maytenus compound on H226, A2058, 786O and HeLa cancer cells and two normal epithelial cell lines, namely, 16HBE and Hecate. Nude mouse xenograft tumor model using H226 and HeLa cells examined the drug's anticancer effect in vivo. Western blot assay studied the possible mechanism. RESULTS: The Maytenus compound indicated obvious ability to against proliferation in four strains of cancer cells, particularly against H226 cells by an IC50 of 85.47±10.06 µg/mL and against HeLa cells by an IC50 of 128.74±17.46 µg/mL. However, it had a low cytotoxicity in human normal epithelial cell lines 16HBE with an IC50 of 4,555.86±25.21 µg/mL and Hecate with an IC50 of 833.56±181.88 µg/mL. The Maytenus compound at the 2.45 g/kg oral dosages inhibited the proliferation of H226 cells and HeLa cells in nude mouse with inhibitory rates of 36.06% and 26.45%, respectively, and no organ toxicity. The Maytenus compound could significantly downregulate the expression of pEGFR, pPI3K, pAKT, pGSK3ß, ß-catenin, and c-MYC and upregulate the protein expression of GSK3ß. CONCLUSIONS: The Maytenus compound has significant anticancer activities against human cancer H226 and HeLa cells both in vitro and in vivo, highlighting it may be an anticancer medicine.

Combination of IFITM1 knockdown and radiotherapy inhibits the growth of oral cancer.

This research aimed to analyze the effect of IFITM1 on the radioresistance of oral neoplasm. Using a multi-group heat map from GSE9716 analysis of the GEO database, IFITM1 was determined to be a relevant radioresistance gene. The TCGA database was analyzed before the expression of IFITM1 was analyzed. IFITM1 expression was quantified by quantitative RT-PCR and immunohistochemistry in 19 paired oral neoplasm cases. The effects of time and dose of radiation on IFITM1 expression level in CAL27 and TSCC1 cell lines were tested by quantitative RT-PCR. Oral neoplasm cells were transfected with siRNA after radiotherapy to disturb IFITM1 expression. After this, the survival rates, cell apoptosis, caspase-3 viability, expression and γ-H2AX were detected using colony formation, flow cytometry, western blot and immunofluorescence, respectively. Western blot was used for STAT1/2/3/p21-related protein and phosphorylation changes. Finally, an in vivo nude mice tumor model was established to verify the effect of IFITM1 on oral neoplasm cells radioresistance. Through microarray analysis, the head and neck neoplasm radioresistance-related gene IFITM1 was found to be overexpressed. IFITM1 overexpression was verified not only using the TCGA database but also in 19 paired cases of oral neoplasm tissues and cells. With increases of dose and time of radiation, the expression of IFITM1 was increased in CAL27 and TSCC1 cell lines. Furthermore, si-IFITM1 may restrain cell proliferation, DNA damage and cell apoptosis in oral neoplasm cell lines. Finally, pSTAT1/2/p21 was found to be upregulated while pSTAT3/p-p21 was downregulated due to IFITM1 inhibition after radiotherapy. The evidence suggested that IFITM1 in combination with radiotherapy can inhibit oral neoplasm cells.

Viral IL-10 down-regulates the "MHC-I antigen processing operon" through the NF-κB signaling pathway in nasopharyngeal carcinoma cells.

The HLA-I antigen processing machinery (APM) plays a crucial role in the anticancer immune response. The loss of surface expression of HLA-I molecules is particularly important as this enables tumor cells to evade recognition and lysis by cytotoxic T-lymphocytes. Transcriptional control of the APM genes is regulated by the nuclear factor kappa B (NF-κB). BCRFl is an Epstein-Barr virus homologue of human IL-10 (hIL-10) and is known as viral IL-10 (vIL-10). vIL-10 shares many immunosuppressive effects with hIL-10 but lacks the immunostimulatory effect of hIL-10. The aim of this study was to assess whether vIL-10 inhibits APM components (TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I) through the NF-κB signaling pathway in nasopharyngeal carcinoma. This work demonstrated that vIL-10 inhibited NF-κB activation by blocking IKK phosphorylation and promoting the expression of IKB. TNF-α treatment led to a strong translocation of NF-κB p65, whereas pretreatment with vIL-10 before TNF-α treatment blocked NF-κB p65 translocation. vIL-10 also inhibited TNF-α-induced DNA-binding of NF-κB p65 in the nucleus. Furthermore, chromatin immunoprecipitation analysis demonstrated that NF-κB p65 could bind to the TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I gene promoters, and after TNF-α stimulation, the down-regulation of TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I transcription by vIL-10 correlated with the suppression of NF-κB in CNE-2 cells. Surprisingly, vIL-10 inhibits only TAP-1 and LMP-7 transcription in CNE-1 cells. Taken together, these results suggest that the inhibition of NF-κB activity may be an important mechanism for vIL-10 suppression of APM (TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I) gene transcription in CNE-2 cells.

Downregulation of expression of transporters associated with antigen processing 1 and 2 and human leukocyte antigen I and its effect on immunity in nasopharyngeal carcinoma patients.

The human leukocyte antigen (HLA)-I and antigen-processing machinery (APM) are crucial in the anti-cancer immune response. The aim of this study was to assess the clinical significance of the APM components [transporters associated with antigen processing (TAP)-1 and -2 and HLA-I] in nasopharyngeal carcinoma (NPC). A total of 58 NPC specimens and 20 healthy specimens used as control were evaluated by semiquantitative immunohistochemistry for three APM components (TAP-1, TAP-2 and HLA-I). The expression of the APM components in NPC was downregulated. CD4+ and CD8+ T cells were measured by flow cytometry and IL-10 was measured by ELISA. The number of CD8+ T cells and the expression of IL-10 were higher and the number of CD4+ T cells was lower in NPC, compared to the controls. The number of CD8+ T cells and the expression of IL-10 were negatively correlated with TAP-1, TAP-2 and HLA-I expression. The clinical phase, lymph node metastasis, distant metastasis, pathological type, TAP-1 expression, TAP-2 expression and HLA-I expression were identified as prognostic factors by the Kaplan-Meier analysis. A multivariate analysis using a Cox regression model indicated that distant metastasis and the downregulation of HLA-I expression were independent unfavorable prognostic factors. In conclusion, the lower expression of HLA-I induced immunosuppression in NPC patients and was associated with a poor prognosis.

[Combined application the siRNA target for Hif-lα and Survivin gene to human NPC CNE-2 cell and its effects on the proliferation and cycle of this cell].

OBJECTIVE: To study the combined transfection of the siRNA target for Hif-lα and Survivin gene to human NPC CNE-2 cell and its effects on the proliferation and cycle of this cell. METHOD: Combined transfection of the siRNA target for Hif-lα and Survivin gene to human NPC CNE-2 cell, these plasmids were respectively transfected into the same cells. Cell proliferation was detected with MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western Blot at the levels of mRNA and protein, respectively. RESULT: MTT showed that CNE-2 cell proliferation in multi-gene plasmid group was more significantly inhibited than a single gene. The expression of mRNA and protein of two different genes were both decreased in HS group, and the interference effect of multiple genes was better than the single-gene(P<0.01). CONCLUSION: HS group could restrain cell proliferation and interference the mRNA and protein expression in nasopharyngeal carcinoma CNE-2 cell, which was better than the other groups.

Clinical significance of joint detection of CD44v6 and CD62P in nasopharyngeal carcinoma.

AIM: The aim of this study was to investigate the expression and clinical prognostic significance of adhesion molecules in nasopharyngeal carcinoma (NPC) tissues and peripheral blood. MATERIALS AND METHODS: Flow cytometry assays for the expression levels of CD44v6 and CD62P protein in peripheral blood and tissues from controls and NPC patients were performed. Clinical and pathological features were reported and analyzed, and a survival study was carried out. RESULTS: The expression of CD44v6 and CD62P in NPC tissues and peripheral blood was higher than that of the control group (p < 0.05). Expression levels in peripheral blood of stage III/IV NPC patients was markedly higher than that of patients in stage I/II (p < 0.05), while it had no statistically significant difference in tissues (p > 0.05). The expression levels of CD44v6 and CD62P in the lymph gland metastasis and distant metastasis group were higher than groups without such metastasis (p < 0.05), and there was no statistical difference in NPC tissues (p > 0.05). The survival rates of NPC groups with low expression in the peripheral blood were higher than those of high-expression groups (p < 0.05). CONCLUSION: Joint detection of CD44v6 and CD62P in the peripheral blood or tissues of NPC patients has diagnostic and prognostic value as a marker of poor clinical outcome.

[Effects of the inhibition and apoptosis by shRNA mediated survivin gene silencing in xenograft tumor of nasopharyngeal carcinoma cell CNE-2].

OBJECTIVE: To observe wether pSIREN-survivin/shRNA can induce the apoptosis and inhibit it's growth of nude mice xenograft tumor of nasopharyngeal carcinoma cell. METHOD: Subcutaneous tumors in athymic mice were induced by inoculation of NPC and divided into the blank group A, the negative group B,the experimental group C randomly. PBS,the negative and positive pSIREN-survivin/shRNA were injected into the subcutaneous tumors poly site. The inhibition ratio was measured by the tumor size calculation after inoculation. The expression of survivin in the xenograft tumor was observed by RT-PCR and immunohistochemistry. The cell apoptosis in tumor tissues was detected by TEM. The liver and kidney function was tested by blood routine test. RESULT: The inhibition ratio of group C and group B was (52. 11 +/- 1. 03)%, (2. 15 0. 11)% respectively, the inhibition rate in expression level of survivin mRNA was (77. 5+/-2. 03) %, (1. 39+/-0. 14) % respectively. The dyeing of survivin was more shallow in group C, the intensity is also less than group B. Nuclear chromatin was deepened, split into pieces, flake, and nuclear membrane was surrounded to edge. Cytoplasmic color and density were deepened, and organelles such as mitochondria was disappeared, the microscopic changes of cellular apoptosis at the earlier stage were watched, the difference of the function in liver and kidney was not statistically in statistical. CONCLUSION: The expression of survivin in xenograft tumor was significantly inhibited by pshRNA-survivin/shRNA,the apoptosis of tumor cells was accelerated and the growth speed of NPC cells in xenograft tumor was retarded. The high expression of nasopharyngeal carcinoma's gene could significantly be silenced by using technology of RNAi, the growth of tumors could be inhibited also. Its a novel treatment that have a good prospect.

[Anatomical study of improved lateral arm free flap].

OBJECTIVE: To study the anatomy and preparation methods of an improved lateral arm free flap (LAFF) for the future clinical application. METHODS: Twenty-two adult upper extremities from cadavers after injected with red latex through common carotid arteries were used. The course, branches, distribution and variations of the blood vessels and nerves of the improved LAFF were observed. The outer diameters of the vessels were measured. RESULTS: The mean length of vascular pedicle of the improved LAFF was (14.85 ± 1.28) cm, significantly more than that (5.46 ± 2.60) of traditional LAFF (t = -8.483, P < 0.001). The mean outer diameters of pedicle arteries and veins in the improved LAFF were (2.24 ± 0.66) mm and (2.22 ± 0.52) mm, significantly more than those (1.15 ± 0.21 and 1.26 ± 0.23) in traditional LAFF (t = -8.690, P < 0.001; t = -15.057, P < 0.001), respectively. CONCLUSION: The improved LAFF has a longer vascular pedicle and larger artery and vein in diameter than conventional LAFF, and is more suitable for the repair of the small and medium-sized defects of the head and neck.

[Treatment and prognosis of sinonasal mucosal melanoma].

OBJECTIVE: To evaluate the treatment and prognosis of sinonasal mucosal melanoma (SMM). METHOD: Clinicopathological data of SMM patients from January 1976 to December 2005 were analyzed retrospectively. Survival analysis was performed and Kaplan-Meier analysis was used to compare the effect of clinicopathological factors on survival using SPSS 18.0 software. A Cox model was applied for multivariate analysis. RESULT: The 3-year and 5-year overall survival (OS) rates of 68 cases of SMM were 36.1% and 29.4%, respectively. The 3-year and 5-year OS of patients who underwent surgery or biotherapy were significantly higher than that of patients who underwent other therapeutic regimens without surgery or without biotherapy, respectively. Multivariate analysis showed the patients with distant metastasis at first present or residual/recurrence had a worse prognosis than that without distant metastasis or residual/recurrence, respectively. Surgery and biotherapy were effective treatments for SMM. CONCLUSION: SMM has a poor prognosis, especially in the patients with distant metastasis or residual/recurrence. Surgery or biotherapy may improve the prognosis of patients with SMM.