Nasopharyngeal Cell Line Research Articles

Detection of cytotoxic effects of carboplatin in human nasopharyngeal carcinoma epithelioid cell line CNE-1 by Raman spectroscopy combined with multivariate statistical analysis

Raman spectroscopy combined with multivariate analysis was used to examine the molecular mechanism of carboplatin toxicity in human nasopharyngeal carcinoma epithelioid cell line CNE-1. Multivariate analysis was performed through principal component analysis combined with linear discriminant analysis, through which a diagnostic algorithm that could discriminate between the control and treatment groups was generated. Compared with the control group (0 μmol/L, 0 hour), the 2 protein Raman peaks (1002 cm−1 and 1659 cm−1) and the 2 deoxyribonucleic acid Raman peaks (1303 cm−1 and 1338 cm−1) in the treatment group decreased with increasing carboplatin concentrations as well as with increasing treatment durations. Using principal component analysis combined with linear discriminant analysis, the discrimination accuracy, sensitivity, and specificity between the control group and the cell groups treated at 12 and 16 μmol/L carboplatin for 24 hours were all 100%, suggesting that 16 μmol/L is suitable as the therapeutic dosing. Using the same method, the discrimination accuracy, sensitivity, and specificity between the control group and the cell groups that were treated with 12 μmol/L carboplatin for 24 and 36 hours were all 100%, indicating that 24 hours could be suitable treatment duration. These results suggest that Raman spectroscopy combined with multivariate statistical analysis could be a useful tool for assessing carboplatin-induced cytotoxicity in human nasopharyngeal carcinoma cells.

Suppression of Nasopharyngeal and Gastric Tumor Growth in a Mouse Model by Antibodies to Epstein-Barr Virus LMP1 Protein.

The study aimed to investigate the antitumor efficacy of anti-LMP1 antibodies in EBV-positive nasopharyngeal and stomach cell lines and xenograft models. The study also examined the NF-κB expression and cell cycle activation of NPC-serum-exosome-associated LMP1. Anti-LMP1 antibody treatment before or during cell implantation prevented tumor growth in nude mice. A small dose of antibodies resulted in complete tumor regression for at least three months after the tumors had grown in size. The consumption of antigen-antibody complexes by tumor cells limited tumor growth. In vitro experiments showed that anti-LMP1 antibodies killed EBV-positive NPC- or GC-derived epithelial cell lines and EBV-positive human B-cell lines but not EBV-negative cell lines. Treatment with anti-LMP1 reduced NF-κB expression in cells. The animal model experiments showed that anti-LMP1 inhibited and prevented NPC- or GC-derived tumor growth. The results suggest that LMP1 antibody immunotherapy could cure nasopharyngeal cancer, EBV-positive gastric carcinoma, and EBV-associated lymphomas. However, further validation of these findings is required through human clinical trials.

Detection of nasopharyngeal cancer cells using the laser tweezer Raman spectroscopy technology.

Nasopharyngeal cancer (NPC), which arises from the nasopharyngeal epithelial lining, is one of the common malignant otorhinolaryngological tumors in China. Due to its insidious anatomical location and highly invasive and metastatic features, it is challenging to detect NPC at early stages. In this work, a rapid laser tweezer Raman spectroscopic (LTRS) system was built and used to trap and characterize single NPC cells. Using LTRS, high-quality Raman signals of the normal nasopharyngeal cell line (NP69) and NPC cells could be successfully obtained. By analysing the Raman peaks, some unique changes were found in components, such as DNA, amide I and amide III, in NPC cells compared with normal cells. In addition, we also used a multivariate statistical algorithm to establish a diagnostic model for identifying NPC cells with an accuracy of 90.0%. These results demonstrate that LTRS in combination with the multivariate statistical analysis is a convenient and high-efficiency cell identification technology, providing a novel and rapid methodology for NPC detection at the single cell level.

Fluorescence lifetime imaging of water-soluble porphyrin in human nasopharyngeal cells under two-photon excitation

A comparative study of the two-photon absorption and emission properties of two free-based porphyrins (H2TMPyP and H2TPPS) and their zinc(II) complexes (ZnTMPyP and ZnTPPS) in three polar solvents has been carried out. The presence of the central zinc(II) ion leads to smaller two-photon absorption cross sections (δTPA), while the introduction of methylpyridinium substituents (N•+–CH3) results in larger ones. The δTPA values of H2TMPyP and ZnTMPyP are affected by both the solvent polarity and the hydrogen bonding between porphyrin and solvent molecules. In this work, the strongest two-photon fluorescence is given by H2TMPyP, which may efficiently get into four kinds of nasopharyngeal cell lines (NP69, SUNE1, CNE2, and CNE1) and prefer to localize in the nucleus. Two-photon fluorescence lifetime images of all the cell lines treated with H2TMPyP consist of a short- (τ1) and a long-lived component (τ2). The τ2 distribution of normal cell line NP69 shows one peak lifetime (∼2.12 ns), while those of the cancerous cell lines (SUNE1, CNE2, and CNE1) have two peak lifetimes (∼2.14 and ∼3.77 ns). The τ1 distribution around 0.68 ns is higher for the well differentiated nasopharyngeal cancerous cell line CNE1 compared with the poorly differentiated ones (SUNE1 and CNE2).

Evaluation of nitroreductase activity in nasopharyngeal carcinoma progression by an activatable two-photon fluorescent probe

Nasopharyngeal carcinoma (NPC) originating from the epithelium cells is the most common malignant tumor of the head and neck. Small-molecule fluorescent probes for early diagnosis of NPC can effectively improve the 5-year survival rate of patients, which makes it become a research hotspot in recent years. Previous studies have suggested the expression levels of NTR in hypoxic tissues or cells and tumors increased relative to the normal state and were positively correlated with the degree of hypoxia. Regarding the mentioned above, we designed a two-photon fluorescent probe NaT-NTR for the detection of NTR in nasopharyngeal cell lines and tissues at different hypoxia levels. NaT-NTR showed high selectivity and sensitivity toward NTR in a complex physiological environment. Furthermore, imaging NTR in different cell lines revealed that the level of intracellular NTR might be positively correlated with the malignancy of nasopharyngeal carcinoma. More importantly, NaT-NTR was successfully applied to detect and image NTR in human nasopharyngeal carcinoma with a penetration depth of 100 µm. On this basis, NaT-NTR might be a powerful chemical tool for the early diagnosis of nasopharyngeal carcinoma.

Abstract 6002: Invasiveness-dependent mechanical plasticity of cancer cells

Abstract Introduction: The capability of living cells to undergo deformations is critical for them to perform different biological functions, such as migration, spreading and endocytosis. Interestingly, recent evidence has also suggested that the mechanical response of cancer cells is intimately related to their pathological state. Physically, a reduced modulus will make it easier for cancer cells to change their morphology when squeezing through tight endothelial cell-cell junctions or pores in the extracellular matrix. However, the deformed cell will fully restore to its original shape if the response is purely elastic or viscoelastic. In this regard, the capability of cells to retain irreversible deformations, and hence effectively “memorize” the characteristics of the physical confinement that they have encountered, could greatly facilitate their subsequent passage through similar barriers, which is a common scenario in tumor cell invasion and metastasis. Here, we developed a method, for the first time to the best of our knowledge, to quantitatively measure the apparent plastic response of tumor cells and examine its correlation with their invasiveness. Materials and Methods: A microfluidic device was designed to deform the cell and then extract its viscoelastic and plastic characteristics from its shape recovery. The chip was casted by molding of silicon wafer which was fabricated using photo etching method. During the test, cells were pushed to squeeze through the deformation channel of the chip. Deformations of the cell and its nucleus were monitored under a fluorescent microscope. Two breast cancer cell lines (MDA-MB-231 and MCF-7), one nasopharyngeal cell line (NP69), and one carcinoma counterpart (HONE-1) were used. To verify the correlation of plastic response with cytoskeleton damage, Latrunculin A was added to block F-actin formation. SYTO fluorescent dye was used for live imaging of the cell nucleus. Results: After being deformed in the narrow channel for a few seconds, cells were released free and recovered the deformation. Interestingly, the highly invasive MDA-MB-231 and HONE-1 cells underwent irreversible deformation even after 10 minutes. However, NP69 and the poorly invasive MCF-7 cells fully recovered to the original round shape in a few minutes. But after blocking the F-actin formation with Latrunculin A, MCF-7 cells retained partial plastic strain. The nucleus elongated when the cell deformed, but fully recovered in 5 minutes. The results indicated the irreversible deformation of cells originated from cytoskeleton damage, but not the nucleus deformation. Conclusions: No apparent irreversible deformation was observed in the less invasive cell lines, suggesting that the plastic response of tumor cells might be correlated with their invasiveness. In addition, it was found that the plastic deformation of highly invasive cancer cells is caused by their cytoskeleton damage rather than plasticity of the nucleus. Citation Format: Xingyu Xia, Zishen Yan, William C. Cho, Yuan Lin. Invasiveness-dependent mechanical plasticity of cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6002.

Investigations on the Role of the MicroRNA-338-5p/Wnt Family Member 2B (WNT2B) Axis in Regulating the Pathogenesis of Nasopharyngeal Carcinoma (NPC).

BackgroundThe involvement of microRNA-338-5p in modulating NPC pathogenesis is still largely unknown, and this study aimed to investigate this issue.MethodsThe expressions of cancer associated genes were determined by Real-Time qPCR and Western Blot, and cell apoptosis was determined by flow cytometer (FCM). CCK-8 assay and colony formation assay were respectively used to determine cell proliferation and colony formation abilities. Transwell assay was used to evaluate cell migration. The expression levels of Ki67 protein in mice tissues were measured by Immunohistochemistry (IHC) assay.ResultsThe present study found that microRNA-338-5p suppressed NPC progression by degrading its downstream target, Wnt family member 2B (WNT2B). Specifically, microRNA-338-5p tended to be low-expressed in NPC tissues and cell lines, compared to the non-tumor nasopharyngeal mucosa tissues and normal nasopharyngeal cell line (NP69). Upregulation of microRNA-338-5p inhibited proliferation, mobility, and epithelial-mesenchymal transition (EMT) in NPC cells in vitro, while silencing of microRNA-338-5p had opposite effects. Consistently, microRNA-338-5p suppressed tumorigenesis of NPC cells in vivo. In addition, microRNA-338-5p targeted WNT2B for degradation and inhibition, and the inhibiting effects of microRNA-338-5p overexpression on NPC development were reversed by upregulating WNT2B.ConclusionsTaken together, we concluded that microRNA-338-5p targeted WNT2B to hinder NPC development.

Epstein-Barr Virus LMP1 Induces Soluble PD-L1 in Nasopharyngeal Carcinoma.

Nasopharyngeal carcinoma (NPC) is an Epstein–Barr virus (EBV)-associated malignancy. The principal oncogene of EBV, latent membrane protein 1 (LMP1), induces the expression of programmed death-ligand 1 (PD-L1), which is an immunosuppressive transmembrane protein and a promising therapeutic target for various malignancies. Recent studies have revealed an association between the level of soluble PD-L1 (sPD-L1) and disease progression. However, the role of sPD-L1 in NPC or its relevance to LMP1 has not been elucidated. This study aimed to examine whether LMP1 induces sPD-L1 in vitro and analyze the clinical relevance of LMP1, PD-L1, and sPD-L1 in NPC patients. Analysis of nasopharyngeal cell lines revealed that LMP1 induces both cellular PD-L1 and sPD-L1. Analysis of biopsy specimens from 32 NPC patients revealed that LMP1 expression was significantly correlated with PD-L1 expression. Finally, the serum sPD-L1 level in NPC patients was higher than that in the controls. Moreover, the sPD-L1 level in the advanced stage was higher than that in the early stage. However, LMP1 expression, PD-L1 expression, and sPD-L1 levels were not associated with prognosis. These results suggest that LMP1 induces both sPD-L1 and PD-L1, which are associated with NPC progression.

Surface modification of gold nanoparticles by cetirizine through surface plasmon resonance and preliminary study of the in vitro cellular cytotoxicity

In this study, gold nanoparticles are synthesized through a conventional redox reaction in water and characterized by means of TEM, DLS, XRD and UV–visible. The average diameter of gold nanoparticles is attained as 18 nm. Then, cetirizine drug is loaded on the as-prepared gold nanoparticles in an aqueous colloidal solution. This investigation shows that the peak of plasmon resonance is sensitive to the concentration of drug, therefore a tentative description is disclosed for this purpose. In addition, a green light source is adopted with the wavelength of plasmon peak of gold nanoparticles to excite the surface plasmons and induce release of the drug molecules. Then, the release profile is studied by UV–visible spectroscopy. It is found that the peak of surface plasmons shows a distinct blue shift, dependent upon the time of light irradiation. Plotting numerical data to the experimental curves for the absorption peaks gives interesting information on the loading/release behavior of cetirizine on the gold nanoparticles. The experimental data were fitted well with an exponential fucnction and we found that ~63% of the drug was released into the solution after about 7 min radiation. These results clearly show that the emitting green light can release the drug molecules, as can be detected by UV–visible absorption experiment. Outputs of this study offer a new insight on the investigation of drug delivery by using excitation of surface plasmon resonance from gold nanoparticles. Finally, the performed cellular cytoxicity and inhibitory dose IC50 of Au@cetirizine nanoparticles on nasopharyngeal carcinoms cell line (C666–1) showed that this nanoparticle has an acceptable cytotoxicity toward the examined cell line and a decrease in the MTT signal was attained compared to the untreated cells.

Long intergenic non-protein coding RNA 02570 promotes nasopharyngeal carcinoma progression by adsorbing microRNA miR-4649-3p thereby upregulating both sterol regulatory element binding protein 1, and fatty acid synthase

ABSTRACT Our previous studies have elucidated a possible connection between long intergenic non-protein coding RNA 2570 (LINC02570) and nasopharyngeal carcinoma (NPC). However, the precise mechanism by which LINC02570 promotes NPC remains unknown. We used quantitative polymerase chain reaction (qPCR) to detect LINC02570 expression in nasopharyngeal cell lines, NPC tissues, and chronic rhinitis tissues. Subcellular LINC02570 localization was confirmed by fluorescence in situ hybridization (FISH). The effects of LINC02570 stable knockdown and overexpression on viabillity, proliferation, migration, and invasion were analyzed using 3-(4,5-Dimethyl-2-Thiazolyl)-2,5-Diphenyl-2-H-Tetrazolium bromide (MTT), a colorimetric focus-formation assay, a wound healing assay, and transwell assays. RNA crosstalk analysis in silico predicted microRNA-4649-3p (miR-4649-3p) binding to LINC02570 or sterol regulatory element binding transcription factor 1 (SREBF1). A dual luciferase reporter assay was used to confirm potential interactions. Sterol regulatory element binding protein 1 (SREBP1) and fatty acid synthase (FASN) expression were detected by western blotting. The results suggest that LINC02570 is upregulated in late clinical stage NPC patients, and promotes NPC progression by adsorbing miR-4649-3p to up-regulate SREBP1 and FASN. This study elucidates a potential chemotherapeutic target involved in lipid metabolism in NPC.

Oridonin inhibits epithelial-mesenchymal transition of human nasopharyngeal carcinoma cells by negatively regulating AKT/STAT3 signaling pathway.

Oridonin, derived from Rabdosia rubescens, has exhibited anticancer activity in a variety of cancers. However, few studies have explored the effect of oridonin (ORI) on migration, invasion and epithelial-mesenchymal transition (EMT) in nasopharyngeal carcinoma. In our study, the results demonstrated that oridonin significantly inhibited migration and invasion of human nasopharyngeal carcinoma CNE-2Z and HNE-1 cell lines, as depicted by wound healing and Transwell assays. In addition, oridonin increased the expression of E-Cadherin while decreased the expressions of vimentin and twist1 at the mRNA and protein levels in a dose-dependent manner. Interestingly, oridonin also decreased cell mobility in nasopharyngeal carcinoma. The subsequent results of western blotting uncovered that the phosphorylation levels of AKT and signal transducer and activator of transcription 3 (STAT3) were decreased upon oridonin treatment. Furthermore, co-treatment with the AKT activator SC-79 attenuated the anti-metastatic effect of oridonin on nasopharyngeal carcinoma and partially abolished the high expression of E-cadherin and the low expression of twist1 mediated by oridonin. In conclusion, the results revealed that oridonin could repress metastatic phenotype and reverse epithelial-mesenchymal transition (EMT) in nasopharyngeal carcinoma by negatively regulating AKT/STAT3 signaling pathway, suggesting that AKT/STAT3 signaling may be the potential therapeutic target of oridonin against nasopharyngeal carcinoma.

Single cell detection using intracellularly-grown-Au-nanoparticle based surface-enhanced Raman scattering spectroscopy for nasopharyngeal cell line classification.

The aim of this study was to evaluate the feasibility of applying intracellularly-grown-Au-nanoparticle (IGAuNP)-based surface-enhanced Raman scattering (SERS) technology to classify two types of nasopharyngeal cancer (NPC) cell lines (CNE2 and CNE1). The IGAuNP technology provides excellent delivery efficiency of Au NPs to the cytoplasm and nucleus, thus leading to an extraordinary enhancement of the Raman signals of cells. Compared with normal Raman scattering (NRS) spectra of cells, IGAuNP-based SERS spectra not only have a high signal-to-noise ratio, but also can detect more characteristic Raman peaks, which can be used to explore more differences when comparing the biochemical components of different nasopharyngeal carcinoma cell lines. Based on the linear discriminant analysis (LDA) and support vector machine (SVM) analysis of SERS spectral data, an exciting result with a diagnostic sensitivity of 100%, specificity of 100%, and accuracy of 100%, could be achieved to differentiate CNE2 and CNE1 cells, which is better than the result obtained by NRS spectroscopy. This exploratory study indicated that the SERS technology based on IGAuNPs in conjunction with multivariate statistical analysis methods has great potential in the identification of nasopharyngeal carcinoma cell lines.

Anticancer and antimicrobial peptides from medicinal plants of Borneo island in Sarawak

The interest in drug discovery from plants-based metabolites has been of interests to researchers, especially for health well-being and for therapeutic reasons. The work described here was to explore the in vitro anticancer and antimicrobial peptides from six traditional medicinal plants commonly used in Sarawak, Malaysian Borneo. Proteins were extracted from plants with a common protein extraction buffer. Evaluation for in vitro anticancer activity was done against normal and carcinomas nasopharyngeal cell line; NP69 and HK-1 respectively by using established MTT microtiter plate assays. Antimicrobial activity was tested against Staphylococcus aureus and Escherichia coli, a Gram-positive and Gram-negative respectively, by agar well diffusion method. The plant extracts were fractionated and in vitro anticancer activity of the fractionated extracts were repeated. Piper sarmentosum and Orthosiphon aristatus extracts did not show any distinguishable inhibition towards nasopharyngeal cell line with the cell viability above 90%. The antimicrobial activity was exhibited by P. sarmentosum, Piper betle and Senna alata extract with more than 1 mm inhibition zones observed. Also, fraction 1 from fractionated P. sarmentosum extracts and fraction 2 from fractionated O. aristatus extracts showed noticeable inhibition towards carcinomas nasopharyngeal cell line with the cell viability below 80%. These results showed that local medicinal plants could be promising sources of natural products with potential anticancer and antimicrobial activity. The results can be used as a guide for plants selection for further pharmacological and phytochemical investigations.

EBV-LMP1 induces APOBEC3s and mitochondrial DNA hypermutation in nasopharyngeal cancer.

An Epstein‐Barr virus (EBV)—encoded latent membrane protein 1 (LMP1) is a principal oncogene that plays a pivotal role in EBV‐associated malignant tumors including nasopharyngeal cancer (NPC). Recent genomic landscape studies revealed that NPC also contained many genomic mutations, suggesting the role of LMP1 as a driver gene for the induction of these genomic mutations. Nonetheless, its exact mechanism has not been investigated. In this study, we report that LMP1 alters the expression profile of APOBEC3s(A3s), host deaminases that introduce consecutive C‐to‐U mutations (hypermutation). In vitro, LMP1 induces APOBEC3B (A3B) and 3F(A3F), in a nasopharyngeal cell line, AdAH. Overexpression of LMP1, A3B, or A3F induces mtDNA hypermutation, which is also detectable from NPC specimens. Expression of LMP1 and A3B in NPC was correlated with neck metastasis. These results provide evidence as to which LMP1 induces A3s and mtDNA hypermutation, and how LMP1 facilitates metastasis is also discussed.

MiR-34c downregulation leads to SOX4 overexpression and cisplatin resistance in nasopharyngeal carcinoma

BackgroundA major cause of disease-related death in nasopharyngeal carcinoma (NPC) is the development of distant metastasis (DM) despite combination chemoradiotherapy treatment. We previously identified and validated a four microRNA (miRNA) signature that is prognostic for DM. In this study, characterization of a key component of this signature, miR-34c, revealed its role in chemotherapy resistance.MethodsTwo hundred forty-six NPC patient biopsy samples were subject to comprehensive miRNA profiling and immunohistochemistry (IHC). Two human normal nasopharyngeal cell lines (immortalized; NP69 and NP460), as well as the NPC cell line C666–1, were used for miR-34c gain-of-function and loss-of-function experiments. Signaling pathways were assessed using quantitative real-time PCR (qRT-PCR) and Western blot. Cell viability was measured using the ATPlite assay.ResultsMiR-34c was downregulated in NPC patient samples, and confirmed in vitro to directly target SOX4, a master regulator of epithelial-to-mesenchymal transition (EMT). MiR-34c downregulation triggered EMT-representative changes in NP69 and NP460 whereby Snail, ZEB1, CDH2, and SOX2 were upregulated, while Claudin-1 and CDH1 were downregulated. Phenotypically, inhibition of miR-34c led to cisplatin resistance, whereas miR-34c over-expression sensitized NPC cells to cisplatin. TGFβ1 decreased miR-34c and increased SOX4 expression in vitro. The TGFβ receptor 1 inhibitor SB431542 reduced SOX4 expression and increased cisplatin sensitivity. Finally, IHC revealed that lower SOX4 expression was associated with improved overall survival in chemotherapy-treated NPC patients.ConclusionmiR-34c is downregulated in NPC. Repression of miR-34c was shown to increase SOX4 expression, which leads to cisplatin resistance, while TGFβ1 was found to repress miR-34c expression. Taken together, our study demonstrates that inhibition of the TGFβ1 pathway could be a strategy to restore cisplatin sensitivity in NPC.

MiR-154-5p Suppresses Cell Invasion and Migration Through Inhibiting KIF14 in Nasopharyngeal Carcinoma.

BackgroundMounting evidence has reported that microRNA-154-5p (miR-154-5p) is involved in the development of multiple cancers, but its function in nasopharyngeal carcinoma (NPC) remains not well investigated.MethodsReal-time quantitative PCR (qRT-PCR) was used to detect miR-154-5p expression in NPC tissues and cells. CCK8, colony formation, wound healing and transwell assays were performed to assess cell proliferation, migration and invasion. Dual-luciferase reporter assays and Western blots were performed to confirm the target gene of miR-154-5p. Rescue experiments were conducted to explore the influence of target gene KIF14 on the functions of miR-154-5p. Xenograft tumor model was conducted to detect the effect of miR-154-5p in vivo.ResultsqRT-PCR results revealed that the expression of miR-154-5p was down-regulated in NPC tissues and cell lines compared to normal nasopharyngeal tissues and cell line. Overexpression of miR-154-5p inhibited cell migration and invasion. However, miR-154-5p had no influence on the proliferation of NPC cells. MiR-154-5p overexpression suppressed xenograft tumor metastasis in vivo. Dual-luciferase reporter analysis identified KIF14 as a target gene of miR-154-5p. Rescue experiments showed that knockdown of KIF14 reversed the effect of inhibiting miR-154-5p expression on NPC cell migration and invasion.ConclusionTaken together, miR-154-5p suppresses tumor migration and invasion by targeting KIF14 in NPC. The newly identified miR-154-5p/KIF14 interaction offers further insights into the progression of NPC, which may represent a novel target for NPC diagnosis and treatment.

Dysregulated NF-κB signal promotes the hub gene PCLAF expression to facilitate nasopharyngeal carcinoma proliferation and metastasis

BackgroundNasopharyngeal carcinoma (NPC) is common in Southern China. The molecular mechanism underlying NPC genesis and progression has been comprehensively investigated, but the key gene (s) or pathway (s) pertaining to NPC are unidentified. MethodsWe explored some key genes and pathways involved in NPC through using meta-analysis of deposited expression of microarray data of NPC. The expression of proliferating cell nuclear antigen clamp associated factor (PCLAF) was determined by real-time PCR and western blots. CCK-8 assay, colony formation assay, transwell migration assay, cell wound healing assay, cell cycle analysis and cell apoptosis were carried out to assess biological behaviors caused by downregulation and overexpression of PCLAF in vitro. CHIP was utilized to determine the direct upstream regulatory transcription factors of PCLAF. ResultsPCLAF was the key gene of NPC, which was significantly up-regulated in NPC cell line compared to the normal nasopharyngeal cell line. Additionally, in vitro assay has demonstrated the down-regulation and overexpression of PCLAF, resulted in significantly suppressed and enhanced NPC proliferation, metastasis and invasion respectively. Furthermore, the up-regulation of PCLAF in NPC is induced by direct binding of dysregulated NF-κB p50/RelB complex to the promoter of PCLAF. ConclusionOur results offer a strategy for re-using the deposited data to find the key genes and pathways involved in pathogenesis of cancer. Our study has provided evidence of supporting the role of PCLAF in NPC genesis and progression.

Inhibition of PTEN Gene Expression by Small Interfering RNA on PI3K/Akt/FoxO3a Signaling Pathway in Human Nasopharyngeal Carcinoma.

The objective of this article is to study the effect of inhibiting phosphatase and tensin homolog deleted chromatosome 10 gene on phosphoinositide 3-kinase/protein kinase B (Akt)/Forkhead homeobox O3a signaling pathway in human nasopharyngeal carcinoma HK-1 cells. Nasopharyngeal carcinoma HK-1 cell lines were divided into PTEN gene interference group (siPTEN), nonspecific small interfering RNA group (siNC), empty vector group (Vector), and no transfection control group (Normal). The mRNA and protein expression levels of PTEN, PI3K, p-Akt, and FoxO3a were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot. Immunofluorescence was used to detect the subcellular localization of PTEN, PI3K, p-Akt, and FoxO3a in HK-1 cells. The proliferation of HK-1 cells was detected by MTT assay, and the apoptosis of HK-1 cells was detected by flow cytometry. Compared with the siNC group, the expression levels of PTEN, FoxO3a messenger RNA, and protein in the siPTEN group were significantly decreased (P < .05), while the expression levels of PI3K, p-Akt messenger RNA, and protein were significantly increased (P < .05). The growth rate of HK-1 cells in the siPTEN group was significantly higher than the siNC group (P < .05), while the apoptosis rate was significantly lower than that of the siNC group (P < .05). Small interfering RNA can inhibit the expression of PTEN in HK-1 cells, and PTEN can participate in the development of NPC by affecting PI3K/Akt/FoxO3a signaling pathway.

Inhibition of nasopharyngeal cells mediated via G2/M cell cycle arrest, blocking PI3K/AKT/mTOR signalling pathway and suppression of cell migration and invasion

Nasopharyngeal carcinoma (NPC) is a less common but destructive cancer. Because of distant metastasis and deleterious side effects, there is persistent need to identify effective treatment options for nasopharyngeal cancer. In the current research work, the anticancer effects of Abietic acid were evaluated against the cisplatin resistant nasopharyngeal cancer cells. MTT (3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide) assay was executed for monitoring cell proliferation rate. Annexin V/propidium iodide and acridine orange/ethidium bromide assays were executed to investigate effects on cell apoptosis. Flow cytometry was employed to observe the effects on different phases of cell cycle. Cancer Cell migration and cell invasion was checked by transwell assays. Western blotting was done to evaluate impact on protein expression. It was found that Abietic acid exerts potent antiproliferative against the nasopharyngeal cancerous cell line and exhibited an IC50 of 20 μM. However the toxic effects of abietic acid in comparison to cancerous cells were seen to be insignificant against the normal cells. The antitumor impact of abietic acid was exerted through apoptosis induction and alteration of apoptosis related proteins (Bax, caspase 3 and 9). Abietic acid also caused (ROS) reactive oxygen species arbitrated changes in the MMP (mitochondrial transmembrane potential) and triggered cell cycle arrest at G2/M phase. Finally, Abietic acid could also inhibit the invasiveness of cancer cells and migration of the NPC cells. Abietic acid could also block the PI3K/AKT/mTOR biochemical signalling cascade in the HNE1 cells which remains a significant therapeutic objective of anticancer drugs. In conclusion, the results indicate that abietic acid might be a promising drug candidate and promising treatment option for nasopharyngeal carcinoma.

M87 FAMILIAL CO-AGGREGATION OF SCHIZOPHRENIA AND EATING DISORDERS IN SWEDEN AND DENMARK

Nasopharyngeal carcinoma (NPC) is a less common but destructive cancer. Because of distant metastasis and deleterious side effects, there is persistent need to identify effective treatment options for nasopharyngeal cancer. In the current research work, the anticancer effects of Abietic acid were evaluated against the cisplatin resistant nasopharyngeal cancer cells. MTT (3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide) assay was executed for monitoring cell proliferation rate. Annexin V/propidium iodide and acridine orange/ethidium bromide assays were executed to investigate effects on cell apoptosis. Flow cytometry was employed to observe the effects on different phases of cell cycle. Cancer Cell migration and cell invasion was checked by transwell assays. Western blotting was done to evaluate impact on protein expression. It was found that Abietic acid exerts potent antiproliferative against the nasopharyngeal cancerous cell line and exhibited an IC50 of 20 μM. However the toxic effects of abietic acid in comparison to cancerous cells were seen to be insignificant against the normal cells. The antitumor impact of abietic acid was exerted through apoptosis induction and alteration of apoptosis related proteins (Bax, caspase 3 and 9). Abietic acid also caused (ROS) reactive oxygen species arbitrated changes in the MMP (mitochondrial transmembrane potential) and triggered cell cycle arrest at G2/M phase. Finally, Abietic acid could also inhibit the invasiveness of cancer cells and migration of the NPC cells. Abietic acid could also block the PI3K/AKT/mTOR biochemical signalling cascade in the HNE1 cells which remains a significant therapeutic objective of anticancer drugs. In conclusion, the results indicate that abietic acid might be a promising drug candidate and promising treatment option for nasopharyngeal carcinoma.