Nasopharyngeal Cells Research Articles

PP-22 promotes autophagy and apoptosis in the nasopharyngeal carcinoma cell line CNE-2 by inducing endoplasmic reticulum stress, downregulating STAT3 signaling, and modulating the MAPK pathway.

Paris polyphylla var. yunnanensis, named Chong Lou, is considered an antitumor substance. In this study, we investigated the effect of PP-22, a monomer purified from P. polyphylla var. yunnanensis, on the nasopharyngeal carcinoma cell line CNE-2 in vitro. The results showed that PP-22 could inhibit the proliferation of CNE-2 cells via the induction of apoptosis, with evidence of the characteristic morphological changes in the apoptosis in the nucleus and an increase in Annexin V-positive cells. In addition, we found that PP-22 could activate the p38 mitogen-activated protein kinase (MAPK) pathway and that this activation was reversed by SB203580, a specific inhibitor of the p38 MAPK pathway. In contrast, PP-22 promoted apoptosis via an intrinsic pathway, including the endoplasmic reticulum stress pathway, in a caspase-dependent manner. A further study showed that PP-22 also induced apoptosis by downregulating the signal transducers and activators of transcription 3 (STAT3) pathway, and the inhibitory effect was also confirmed by STAT3 small interfering RNA. In addition, PP-22 could promote autophagy by inhibiting the extracellular regulated protein kinases (ERK) pathway. And autophagy plays a protective role against apoptosis. Together, these data show that PP-22 promotes autophagy and apoptosis in the nasopharyngeal carcinoma CNE-2 cell line.

Targeted regulationof STAT3 by miR-29a in mediating Taxol resistance of nasopharyngeal carcinoma cell line CNE-1.

STAT3 is an important molecule in Janus kinase (JAK) signal transducer and activator of transcription (STAT) signal pathway, and facilitates expression of various oncogenic genes such as Bcl-2, thus is correlated with tumor onset, progression and drug resistance. MiR-29a down-regulation is associated with the pathogenesis of nasopharyngeal carcinoma. Bioinformatics analysis demonstrated a complementary binding between miR-29a and 3'-UTR of STAT3. This study aims to investigate the role of miR-29a in regulating STAT3, as well as in Taxol resistance of nasopharyngeal carcinoma CNE-1 cells. Dual luciferase reporter gene assay showed a regulatory relationship between miR-29a and STAT3. Rhodamine 123 repository in CNE-1 and CNE1/Taxol drug resistant cells was measured together with the expression of miR-29a, STAT3, and p-STAT3. Flow cytometry was used to measure cell apoptosis and PCNA expression under Taxol treatment. CNE-1/Taxol cells were treated with miR-29a mimic and or si-STAT3, followed by measuring the expression of miR-29a, STAT3, and p-STAT3 and cell apoptosis. CCK-8 assay was performed to evaluate cell proliferation. MiR-29a inhibited STAT3 expression. Significantly lower Rhodamine 123 repository, miR-29a expression and apoptosis and higher expression of STAT3, p-STAT3 and PCNA were observed in CNE-2/ Taxol cells than those in CNE-1 cells. Transfection of miR-29a mimic and/or si-STAT3 decreased STAT3, p-STAT3 and PCNA expression, inhibited proliferation and promoted cell apoptosis. MiR-29a down-regulation is correlated with drug resistance of nasopharyngeal carcinoma cell line CNE-1 and MiR-29a up-regulation decreases Taxol resistance of nasopharyngeal carcinoma CNE-1 cells possibly via inhibiting STAT3 and Bcl-2 expression.

Association between high survivin expression and late clinical stage of nasopharyngeal non-keratinizing squamous cell carcinoma

Association between high survivin expression and late clinical stage of nasopharyngeal non-keratinizing squamous cell carcinoma

Berberine Hydrochloride-Loaded Chitosan Nanoparticles Effectively Targets and Suppresses Human Nasopharyngeal Carcinoma.

Nasopharyngeal carcinoma (NPC) is a common epithelial malignancy that occurs in the nasopharynx and it is one of the high incidences of malignant tumors in China. As previous reports, berberine hydrochloride (BH) possesses a repressive effect on the proliferation of various cancer types. But the application of BH was hampered for a long time due to its hydrophobic properties, along with poor stability and bioavailability. In this study, folate acid modified chitosan nanoparticles loaded berberine hydrochloride (BH/FA-CTS NPs) were prepared by the ionic cross-linking technique, the physicochemical properties and the influence of BH/FA-CTS NPs on proliferation, migration and apoptosis in human nasopharyngeal carcinoma CNE-1 cells were studied in vitro and in vivo. Results of scanning electron microscopy (SEM) images showed that BH/FA-CTS NPs were sphere and the diameter was 258.2±9.1 nm. The drug loading and encapsulation efficiency were 8.17±1.12% and 87.73±4.21%, respectively. The drug release kinetics exhibited a slower and sustained release over a period of 48 h. Moreover, results of MTT assay and scratch assay displayed that BH/FA-CTS NPs inhibited proliferation and migration of CNE-1 cells remarkably. BH/FA-CTS NPs promoted apoptosis and necrosis of CNE-1 cells. And BH/FA-CTS NPs displayed distinguished higher tumor inhibition than control group, free BH and BH/CTS NPs in vivo. Therefore, as a nanocomposite, BH/FA-CTS NPs provide a new method and option for the treatment of nasopharyngeal carcinoma patients.

X线诱导人鼻咽鳞状细胞癌CNE1细胞株MDR-1及P-gp的表达研究

Objective To examine the expressions of multidrug resistance gene (MDR)-1 and P-glycoprotein (P-gp) in nasopharyngeal squamous cell carcinoma CNE1 cell line before and after X-ray exposure. Methods CNE1 cells were exposed to X-ray. After the irradiation, the CNE1 cells were cultured for 24 hours and tested. The mRNA expressions of MDR-1 in CNE1 cells were measured by semi-quantitative real-time polymerase chain reaction (RT-PCR) before and after X-ray exposure, and the protein expressions of P-gp in CNE1 cells were detected by Western blotting. The protein expressions of P-gp in CNE1 cells were observed by confocal microscope before and after X-ray exposure. Results The results of RT-PCR showed that the absorbance (A) values of MDR-1 mRNA in CNE1 cells were 0.17±0.01 and 0.34±0.03 before and after irradiation, and the difference was statistically significant (t=16.541, P<0.001). The results of Western blotting showed that the A values of P-gp protein in CNE1 cells were 0.02±0.01 and 0.04±0.01, and the difference was statistically significant (t=4.612, P=0.016). The green fluorescence intensity of P-gp protein in CNE1 cells after X-ray irradiation was higher than that before X-ray irradiation by confocal microscope. Conclusion X-ray irradiation can cause the high expressions of MDR-1 and P-gp in CNE1 cells, suggesting that X-ray irradiation can induce the occurrence of multidrug resistance in CNE1 cells. Key words: Nasopharyngeal neoplasms; CNE1 cells; Drug resistance, multiple; X-ray irradiation

Long chain non-coding RNA MALAT-1 gene knockdown inhibits growth and migration and promotes apoptosis of human laryngeal squamous cell carcinoma Hep-2 cells in vitro

To investigate the effect of knocking down long chain non-coding RNA MALAT-1 gene on the biologicalbehaviors of human laryngeal squamous cell carcinoma Hep-2 cells. With immortalized nasopharyngeal epithelial(NPE) cell line NP-69 as the reference, MALAT1 expression in FaDu, Hep-2 and nasopharyngeal carcinoma CNE-2Z cells weredetected using real-time PCR. Hep-2 cells were transfected with shmalat1 lentivirus and the expression of MALAT1 wasdetected. MTT assay, flow cytometry, Transwell assay and M Atrigel invasiveness test were used to evaluate the effect ofMALAT-1 knockdown on the proliferation, cell cycle, cell apoptosis, migration, and invasiveness of Hep-2 cells. Compared with NP-69 cells, Hep-2 cells, FaDu cells, and CNE-2Z cells all showed significantly increased MALAT-1expression. In Hep-2 cells, knockdown of MALAT-1 significantly inhibited the cell proliferation, increased the cell percentagein S phase (P < 0.01), decreased the cell percentage in G2/M phase (P < 0.01), and attenuated the migration and invasiveness of thecells. MALAT-1 is over-expressed in laryngeal squamous cell carcinoma, and knocking down MALAT-1 gene cansignificantly suppress the proliferation, invasion and migration and promotes apoptosis of the cancer cells.

The CRISPR/Cas system in Neisseria meningitidis affects bacterial adhesion to human nasopharyngeal epithelial cells

ABSTRACTNeisseria meningitidis, a commensal β-proteobacterium of the human nasopharynx, constitutes a worldwide leading cause of sepsis and epidemic meningitis. A recent genome-wide association study suggested an association of its type II-C CRISPR/Cas system with carriage and thus less invasive lineages. Here, we show that knock-out strains lacking the Cas9 protein are impaired in the adhesion to human nasopharyngeal cells which constitutes a central step in the pathogenesis of invasive meningococcal disease. Transcriptome sequencing data further suggest that meningococcal Cas9 does not affect the expression of surface adhesins but rather exerts its effect on cell adhesion in an indirect manner. Consequently, we speculate that the meningococcal CRISPR/Cas system exerts novel functions beyond its established role in defence against foreign DNA.

Serum- and glucocorticoid-inducible kinase 3 is a potential oncogene in nasopharyngeal carcinoma

IntroductionSerum- and glucocorticoid-inducible kinase 3, a serine/threonine kinase that functions downstream of the PI3K signaling pathway, plays a critical role in neoplastic processes. It is expressed by various tumors and contributes to carcinogenesis. ObjectiveThe objective was to investigate serum- and glucocorticoid-inducible kinase 3 expression in nasopharyngeal carcinoma, to study the anti-tumor effects of serum- and glucocorticoid-inducible kinase 3 shRNA by inhibiting its expression in nasopharyngeal carcinoma cells and to discuss the potential implications of our findings. MethodsSerum- and glucocorticoid-inducible kinase 3 protein expression in nasopharyngeal carcinoma cell lines (CNE-1, CNE-2, HNE-1, HONE-1, and SUNE-1) and the human immortalized nasopharyngeal epithelium cell line NP69 were assayed by western blotting. Serum- and glucocorticoid-inducible kinase 3 expression in 42 paraffin-embedded nasopharyngeal carcinoma tissues were performed by immunohistochemistry. MTT assay, flow cytometry, and scratch tests were performed after CNE-2 cells were transfected with the best serum- and glucocorticoid-inducible kinase 3 shRNA plasmid selected by western blotting using lipofectamine to study its effect on cell proliferation, apoptosis, and migration. ResultsSerum- and glucocorticoid-inducible kinase 3 was overexpressed in human nasopharyngeal carcinoma tissues and cells. Serum- and glucocorticoid-inducible kinase 3 expression decreased markedly after CNE-2 cells were transfected with the serum- and glucocorticoid-inducible kinase 3 shRNA, leading to strong inhibition of cell proliferation and migration. In addition, the apoptosis rate increased in CNE-2 cells after serum- and glucocorticoid-inducible kinase 3 knockdown. ConclusionSerum- and glucocorticoid-inducible kinase 3 expression was more frequently observed as the nasopharyngeal epithelium progresses from normal tissue to carcinoma. This suggests that serum- and glucocorticoid-inducible kinase 3 contributes to the multistep process of NPC carcinogenesis. Serum- and glucocorticoid-inducible kinase 3 represents a target for nasopharyngeal carcinoma therapy, and a basis exists for the further investigation of this adjuvant treatment modality for nasopharyngeal carcinoma.

The apoptotic effect of Zoledronic acid on the nasopharyngeal carcinoma cells via ROS mediated chloride channel activation.

Zoledronic acid (ZA), a third-generation bisphosphonate, has been applied for treatment of bone metastases caused by malignant tumors. Recent studies have found its anti-cancer effects on various tumor cells. One of the mechanisms of anti-cancer effects of ZA is induction of apoptosis. However, the mechanisms of ZA-induced apoptosis in tumor cells have not been clarified clearly. In this study, we investigated the roles of chloride channels in ZA-induced apoptosis in nasopharyngeal carcinoma CNE-2Z cells. Apoptosis and chloride current were induced by ZA and suppressed by chloride channel blockers. After the knockdown of ClC-3 expression by ClC-3 siRNA, ZA-induced chloride current and apoptosis were significantly suppressed, indicating that the chloride channel participated in ZA-induced apoptosis may be ClC-3. When reactive oxygen species (ROS) generation was inhibited by the antioxidant N-acetyl-L-cysteine (L-NAC), ZA-induced apoptosis and chloride current were blocked accordingly, suggesting that ZA induces apoptosis through promoting ROS production and subsequently activating chloride channel.

Abstract 1108: M2-polarized macrophages increase invasiveness of EBV-associated nasopharyngeal carcinoma by inducing invadopodia formation

Abstract Cancer metastasis is a major cause of mortality, accounting for approximately 90% of cancer related deaths. Emerging evidences suggest that tumour microenvironment plays an indispensable role in cancer metastasis. Recruitment of innate and adaptive immune cells to the tumour results in chronic inflammation which favours tumour growth and progression. Particularly, tumour associated macrophages (TAMs) are found to promote the initiation, growth, and metastasis in several types of cancers. Nasopharyngeal carcinoma (NPC) is one of the highly invasive and metastatic cancers and is closely associated with EBV infection. We postulate that TAMs may increase the invasiveness of NPC by enhancing the formation of invadopodia, which are membranous actin-rich protrusions with digestive ability of extra-cellular matrix. In this study, monocyte THP-1 cells were polarized to M2-macrophages, and co-cultured with nasopharyngeal cells in a contained chamber-insert. The stimulated nasopharyngeal cells then formed massive numbers of invadopodia and digested larger area of gelatin compared with the unstimulated counterparts. A cytokine antibody array was used to identify the upregulated cytokines in the co-culture medium of the M2-macrohphages and the NPC cells. Tumour necrosis factor alpha (TNFα) was revealed to be the responsible cytokine to induce the formation of invadopodia and digestion of gelatin. Besides, western blot analysis identified that several actin-associated proteins such as Src, Erk, and cortactin were involved in the TNFα-mediated invadopodia formation. We also found that the expression of latent membrane protein 1 (LMP1), an EBV protein that has similar structure to TNF receptor was elevated in NPC cells after co-culturing with the M2-polarized macrophages. Through transient transfection and stable expression of LMP1 in the NPC cells, LMP1 could also promote the formation of invadopodia through cdc42 activation. Taken together, these data indicate macrophages can promote the invasiveness of EBV-infected NPC cells by enhancing their ability in forming invadopodia through an activated TNFα signaling axis and LMP1 upregulation. Citation Format: Wilson Wing Chung Tang, Chan Ping You, Gareth E. Jones, George Sai-Wah Tsao, Anna Chi Man Tsang. M2-polarized macrophages increase invasiveness of EBV-associated nasopharyngeal carcinoma by inducing invadopodia formation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1108.

Abstract 4563: PD-L1 expression associated with treatment responses in colorectal cancer patients with XELOX/FOLFOX chemotherapy: Potential of checkpoint blockage and natural killer cell-based immunotherapy

Abstract Chemo-resistance is a major obstacle in human cancer treatment. Immunotherapies including checkpoint blockage and cell-based therapy have been clinically validated for cancer treatment. The present study aimed to investigate the role of PD-L1 and potential of immunotherapy for chemo-resistant cancers. In a pilot study, we have examined the expression of PD-L1 in colorectal cancer (CRC) patients who received XELOX (capecitabine plus oxaliplatin) or FOLFOX (fluorouracil plus oxaliplatin) chemotherapy. Detection rate of PD-L1 expression by immunohistochemistry was higher in CRC patients who demonstrated poor response (5/10, 50%) compared to good responders (0/5, 0%) on XELOX/FOLFOX chemotherapy. Chemo-resistant CRC cell lines (HT29 and HCT116) were used to examine the role of PD-L1. Cells were plated out, selected under fluorouracil and oxaliplatin in serial, and referred as fluorouracil-oxaliplatin-resistant (FOR) cells. Half maximal inhibitory concentration (IC50) for respective drug was determined by MTT assay. HT29 FOR, compared to parental cells, showed chemo-resistance (359.4 and 10.1 fold increase to fluorouracil and oxaliplatin respectively) and similarly for HCT116 FOR (8.0 and 9.0 fold increase respectively). Hepatocellular carcinoma Hep3B and nasopharyngeal carcinoma C666-1 cells that have been selected for fluorouracil chemo-resistant (FR) were included. All the chemo-resistant cell lines showed increased expression of PD-L1 by real-time qPCR compared to their respective parental cells (HT29 FOR: 6.1 fold; HCT116 FOR: 1.9 fold; Hep3B FR: 3.1 fold; C666-1 FR: 4.4 fold). To explore the potential of immunotherapy, natural killer (NK) cells were isolated from peripheral blood PBMC of healthy donors. NK cells were expanded and activated ex vivo in stem cell growth medium supplemented with interleukins. NK co-culture assay was performed and apoptosis of cancer cells examined by flow cytometry. Parental cancer cells were sensitive to cytotoxic killing by NK cells compared to control no NK treatment (36.0% vs 2.8% apoptotic cells, P=0.001), and PD1 antibody (Nivolumab) treatment promoted NK-mediated cytotoxicity (52.9%, P=0.034). Notably, chemo-resistant cancer cells were similarly sensitive to cytotoxic killing by NK cells compared to control (27.1% vs 3.4%, P=0.002), and PD1 antibody (Nivolumab) treatment further enhanced NK-mediated cytotoxicity (52.8%, P=0.013). In summary, PD-L1 expression was associated with poor response for XELOX/FOLFOX chemotherapy in CRC patients. Chemo-resistant cancer cells showed elevated PD-L1 expression, nonetheless, susceptible to NK-mediated cytotoxicity and checkpoint blockage by PD1 antibody augmented the treatment response. Further studies are warranted to examine immunotherapy for chemo-resistant cancers. Citation Format: Elaine H.L. SIU, Simon S.M. NG, Anthony W.H. CHAN, James Y.W. LAU, Kwok Wai LO, Charing C.N. CHONG, Paul B.S. LAI, Stephen L. CHAN, Marcus H.N. LAW, Linda W.C. NG, S.T. CHEUNG. PD-L1 expression associated with treatment responses in colorectal cancer patients with XELOX/FOLFOX chemotherapy: Potential of checkpoint blockage and natural killer cell-based immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4563.

Abstract 1913: Investigation of targeting SKP2 to improve the treatment response in glioma

Abstract Glioma is the most frequent tumor in central nervous system (CNS), constituting about 30% of CNS tumors and 80% of malignant brain tumors. The current standard strategy for glioma treatment is surgery plus adjuvant chemotherapy and concurrent radiotherapy. However, the prognosis of glioma is still poor. It has been proved that the existence of glioma stem cells and the timely repair of DNA damages are the main causes of treatment failure. Skp2 (S-phase kinase associated protein-2), a member of F-box proteins, forms the Skp2 SCF complex with Skp1, Cullin-1, and Rbx1, serves as one of the important E3-ligases in the process of ubiquitination, and is responsible for substrate recognition. Skp2 is overexpressed in a variety of human cancers and promotes cancer progression by inducing p27 degradation. Importantly, Skp2 deficiency profoundly restricts cancer progression in multiple genetic mouse tumor models. Our group has demonstrated that Skp2 is involved in the regulation of pool size and self-renewal ability of hematopoietic stem cells, as well as breast cancer stem cells and nasopharyngeal carcinoma stem cell-like cells. However, the exact role of Skp2 in the regulation of glioma stem cells and DNA damage repair in glioma has not been elucidated yet. We proposed that targeting Skp2 could promote radio- and chemotherapy sensitivity through suppressing function of glioma stem cells. We first detected the expression of SKP2 in glioma tumor tissues and found that SKP2 was highly expressed in glioma and related to the prognosis of glioma patients. We also adopted gene knockdown methods and inhibitor pretreatment with small molecules in order to suppress the level of Skp2, and then checked the sphere formation and self-renewal ability of glioma stem cell upon radio/chemo treatments. In this way we evaluated the role of Skp2 in repressing the glioma cell proliferation and subsequently triggering apoptosis. Our project will clarify the novel mechanisms about how Skp2 is involved in the glioma radio- and chemotherapy resistance, which will provide a potential target to improve the radio/chemo therapeutic effect through targeting one single target. Citation Format: Jing Wang, Hong-Kai Su, Zhong-Ping Chen, Yue Qu, Hai-Ping Cai. Investigation of targeting SKP2 to improve the treatment response in glioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1913.

Synthesis of cell penetrating peptide decorated magnetic nanoparticles loading cisplatin for nasopharyngeal cancer therapy

Objective:To synthesize cisplatin loaded and cell penetrating peptide TAT decorated magnetic nanoparticles and to observe the inhibiting effect in vitro on nasopharyngeal cancer therapy.Method：The aldehyde sodium alginate coated magnetic nanoparticles (ASA-MNPs) was prepared as the drug delivery system, which was covalently attached by PEGylation TAT (TAT-ASA-MNPs) via condensation of aldehyde with amino group and then coordinated with cisplatin (TAT-ASA-MNPs@CDDP). The complex was characterized by H-NMR and FT-IR. The cell penetrating ability and biocompatibility were observed by means of fluorescent tags. The inhibited effect on nasopharyngeal cancer CNE-2 cells was measured by cellular toxicity research and flow cytometry.Result：The H NMR and FT-IR of TAT-ASA-MNPs exhibited the characteristic peaks of TAT, PEG as well as ASA. The dynamic light scattering showed the hydrodynamic diameter of the complex was（145.9±1.5）nm. Zeta potential was（-21.66±1.24）mV and the drug loading rate was（25.03±3.05）%. Fluorescent labeling assay revealed that FITC marked TATASAMNPs was quickly taken up by CNE-2 cells. Cytotoxicity experiment on 293T cells displayed high survival rate (>70%) after cultured for 72h. Negative hemagglutination reflected decent biocompatibility. In vitro cytotoxicity test and cell apoptosis assay exhibited obvious inhibition on CNE-2 cell with TATASAMNPs@CDDP at low concentration of cisplatin compared to ASA-MNPs@CDDP (P<0.05).Conclusion：TAT-ASA-MNPs showed decent biocompatibility while distinctly inhibit CNE-2 cells in vitro study.

Abstract 4694: The contribution of Epstein-Barr virus to the impaired T-cell responses in nasopharyngeal carcinoma

Abstract Undifferentiated nasopharyngeal carcinoma (NPC) is a geographically skewed disease in which more than 80% cases are from southern China and Southeast Asia. NPC is consistently associated with Epstein-Barr virus (EBV) infection and characterized by a prominent lymphocyte infiltration. The consistent expression of EBV proteins in NPC cells has led to several clinical trials of adoptive T-cell therapy or vaccination to boost the immune response to EBV antigens. However, these treatments yield relatively modest clinical benefit, which may be due to varying degrees of immunosuppression in the tumor microenvironment. The lack of model systems to analyze the ability of therapeutically relevant T cells to recognize and kill NPC cells represents a fundamental limitation in our attempts to harness the immune system for clinical benefit. We used three NPC cell lines as targets in T-cell assays, together with a panel of established HLA-matched EBV-specific CD8 and CD4 T-cell clones. T-cell activity and killing were measured by IFN-γ ELISA and outgrowth assays, respectively. We demonstrate, for the first time, that NPC cells can be recognized and killed by EBV-specific T-cell clones in vitro, which suggests that tumor-derived factors inhibit such responses in vivo. These observations are consistent with the previous reported functional inactivation of EBV-specific cytotoxic T lymphocytes (CTL) in NPC patients. We hypothesized that EBV infection of NPC cells induces the secretion of immunosuppressive factors. The effect of conditioned media derived from an EBV-infected premalignant nasopharyngeal cell line, NP460htert, on T-cell activity was examined by treating several clones of EBV-specific CTLs in vitro. The conditioned media inhibited the secretion of IFN-γ from peptide-stimulated CTLs, indicating that secreted factors that could suppress CTL functions are induced following EBV infection in NP460htert cells. RNA sequencing analyses on NP460hTert and three NPC cell lines following EBV infection revealed a significant enrichment of genes whose proteins are located at extracellular region/matrix, including a number of immunomodulators. Our results suggest that EBV infection might induce the production of soluble molecules, which in turn suppress the antitumor T-cell responses. In conclusion, we have developed a system with which to study NPC-specific immune responses and shown that soluble factors induced by EBV infection can suppress T-cell activity. These model systems will be critical in the future to examine the effects of checkpoint inhibitors, secreted factors from cancer-associated fibroblasts or NPC cells upon T-cell activity, which will ultimately enhance the success of EBV-targeted immunotherapy strategies. Citation Format: Hui Min Lee, Tracey Haigh, George SW Tsao, Paul G. Murray, Graham Taylor, Ian C. Paterson, Lee Fah Yap. The contribution of Epstein-Barr virus to the impaired T-cell responses in nasopharyngeal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4694.

Lanyamycin, a macrolide antibiotic from Sorangium cellulosum, strain Soce 481 (Myxobacteria).

Lanyamycin (1/2), a secondary metabolite occurring as two epimers, was isolated from the myxobacterium Sorangium cellulosum, strain Soce 481. The structures of both epimers were elucidated from HRESIMS and 1D and 2D NMR data and the relative configuration of their macrolactone ring was assigned based on NOE and vicinal 1H NMR coupling constants and by calculation of a 3D model. Lanyamycin inhibited HCV infection into mammalian liver cells with an IC50 value of 11.8 µM, and exhibited a moderate cytotoxic activity against the mouse fibroblast cell line L929 and the human nasopharyngeal cell line KB3 with IC50 values of 3.1 and 1.5 μM, respectively, and also suppressed the growth of the Gram-positive bacterium Micrococcus luteus.

Cofilin-2 Acts as a Marker for Predicting Radiotherapy Response and Is a Potential Therapeutic Target in Nasopharyngeal Carcinoma.

BackgroundThe purpose of this study was to determine whether cofilin-2 could serve as a protein marker for predicting radiotherapy response and as a potential therapeutic target in nasopharyngeal carcinoma (NPC).Material/MethodsCofilin-2 protein levels in serum and tissue samples from patients with NPC were assessed by sandwich ELISA and IHC. In vitro, cofilin-2 levels in CNE-2R cells were significantly higher than those of CNE-2 cells. Meanwhile, CNE-2R cells were silenced for cofilin-2 to obtain a stable cofilin-2-RNAi-LV3 cell line. Then, cell proliferation, radiosensitivity, invasion and migration abilities, cell cycle, and apoptosis were evaluated by Cell Counting Kit 8 assay (CCK-8), flow cytometry (FCM), clone formation assay, and in vitro.ResultsThe secreted levels of the cofilin-2 protein in radioresistant NPC patients were significantly higher than those of radiosensitive cases. After cofilin-2 knockdown in nasopharyngeal carcinoma CNE-2R cells, proliferation was decreased, while apoptosis and radiosensitivity were enhanced; cell cycle distribution was altered, and the transplanted tumors in nude mice grew significantly less.ConclusionsOverall, our findings suggest that cofilin-2 acts as a marker for predicting radiotherapy response and is a potential therapeutic target in nasopharyngeal carcinoma.

Association Between Dose and Duration of Cisplatin Exposure with Cytotoxicity Effect on Nasopharyngeal Carcinoma Stem Cell.

Nasopharyngeal carcinoma (NPC) is ranked 6th of malignant tumors in Indonesia. To analyze the correlation of dose and duration of cisplatin exposure with cytotoxic effects on nasopharyngeal carcinoma stem cells. The biopsy NPC tissue was cultured and processed to obtain NPC stem cells to be treated with cisplatin different doses and durations (24 and 48h). The number of dead cells after exposure will be calculated using a hemocytometer. Death stem cell density of NPC at exposure of 2μg/ml cisplatin dose was 81.37%, while the smallest death cell density a dose of 0.05μg/ml after a 72-h observation was 21.3%. The coefficient correlation 0.827 and value p = 0.000 (p < 0.05). The analysis of the correlation between cisplatin exposure duration and death cell was also significant with the correlation coefficient -0.357 and the value p = 0.001 (p < 0.05). There was a correlation between the increased dose of cisplatin with the cytotoxicity effects on NPC stem cell.

Submandibular Gland Transfer: A Potential Imaging Pitfall.

The Seikaly and Jha submandibular gland transfer surgery is performed to facilitate gland shielding during radiation therapy for head and neck tumors to circumvent radiation-induced xerostomia. It results in an asymmetric postsurgical appearance of the submandibular and submental spaces. Our purpose was to characterize the morphologic and enhancement characteristics of the transferred submandibular gland and identify potential pitfalls in postoperative radiologic interpretation. This retrospective study identified patients with head and neck cancer who had undergone the submandibular gland transfer procedure at our institution. Chart reviews were performed to identify relevant oncologic histories and therapies. CT and MR neck imaging was reviewed to characterize morphologic and enhancement characteristics of the pre- and postoperative submandibular glands, as well as interpretive accuracy. Eleven patients with oropharyngeal and nasopharyngeal squamous cell carcinomas who underwent submandibular gland transfer were identified. The transferred glands were significantly lengthened in the anteroposterior dimension compared with contralateral glands (P < .001) and displaced anteriorly and inferiorly within the submandibular and submental spaces. Enhancement patterns of the transferred submandibular glands varied, depending on the time of imaging relative to the operation and radiation therapy. Submandibular gland transfer was acknowledged in the postoperative report in 7/11 cases. Errors in interpretation were present in 2/11 reports. After the submandibular gland transfer procedure, the submandibular and submental spaces lose their symmetric appearances as the transferred submandibular glands become lengthened and located more anteriorly and inferiorly, with variable enhancement characteristics. Familiarity with the postsurgical appearance of the transferred submandibular glands is key to accurate imaging interpretation.

Quantitative analysis of γ-H2AX foci formation and dynamic changes in DNA double-strand breaks induced by X-ray radiation

Objective To quantitatively compare the γ-H2AX foci formation between DNA-PKcs+ /+ and DNA-PKcs-/-mouse embryonic fibroblast (MEF) cells, and to investigate the dynamic changes in DNA double-strand breaks (DSBs) in human nasopharyngeal carcinoma SUNE-1 cells exposed to X-ray radiation. Methods The expression of DNA-PKcs was determined by Western blot. The γ-H2AX foci formation induced by 5 Gy X-ray radiation was detected by cell immunofluorescence. The ImageJ software was used to quantitatively analyze the γ-H2AX foci formation. Results The expression of DNA-PKcs was silenced in DNA-PKcs-/-MEF cells and normal in DNA-PKcs+ /+ MEF cells. According to the dynamic analyses of the numbers of γ-H2AX foci/cell and γ-H2AX foci/mm2, a similar tendency was observed in DSB formation in DNA-PKcs+ /+ MEF cells, DNA-PKcs-/-MEF cells, and SUNE-1 cells exposed to X-ray radiation. A large number of γ-H2AX foci formed at 0.5-1.0 h after radiation. DSBs were repaired at 6 h after radiation in DNA-PKcs+ /+ MEF cells and 24 h after radiation in DNA-PKcs-/-MEF cells and SUNE-1 cells. The peak values of γ-H2AX foci/cell and γ-H2AX foci/mm2 were observed at 1.0 and 0.5 h after radiation, respectively. Compared with DNA-PKcs+ /+ MEF cells, DNA-PKcs-/-MEF cells had different numbers of γ-H2AX foci/cell at 0.5, 1.0, 3.0, 6.0, and 12.0 h after radiation, as well as different numbers of γ-H2AX foci/mm2 at 3.0, 6.0, and 12.0 h after radiation. Conclusions Quantitative measurement of the number of γ-H2AX foci/cell or γ-H2AX foci/mm2 by cell immunofluorescence provides new insights into the quantitative and dynamic study of DSB damage and repair. Key words: Non-homologous terminal connection; DNA dependent protein kinase catalytic subunit; γH2AX foci formation; SUNE-1 cell line

Induction of apoptosis by Streptomyces strain CH54-4 extract through activation of caspase-3 in human nasopharyngeal cells

Induction of apoptosis by Streptomyces strain CH54-4 extract through activation of caspase-3 in human nasopharyngeal cells