epidermal-growth-factor and Nasopharyngeal-Neoplasms

Oncogenic viruses may have a significant impact on the therapeutic management of several malignancies besides their well-known role in tumor pathogenesis. Epstein-Barr virus (EBV) induces neoplastic transformation of epithelial cells of the nasopharynx by various molecular mechanisms mostly involving activation of oncogenes and inactivation of tumor-suppressor genes. EBV infection can also induce the expression of several immunogenic peptides on the plasma membrane of the infected cells. Importantly, these virus-related antigens may be used as targets for antitumor immunotherapy-based treatment strategies. Two different immunotherapy strategies, namely adoptive and active immunotherapy, have been developed and strongly improved in the recent years. Furthermore, EBV infection may influence the use of targeted therapies for nasopharyngeal carcinoma (NPC) considering that the presence of EBV can induce important modifications in cell signaling. As an example, latent membrane protein type 1 is a viral transmembrane protein mainly involved in the cancerogenesis process, which can also mediate overexpression of the epidermal growth factor receptor (EGFR) in NPC cells, rendering them more sensitive to anti-EGFR therapy. Finally, EBV may induce epigenetic changes in the infected cells, such as DNA hypermethylation and histone deacetylation, that can sustain tumor growth and can thus be considered potential targets for novel therapies. In conclusion, EBV infection can modify important biological features of NPC cells, rendering them more vulnerable to both immunotherapy and targeted therapy.

Topics: Antigens, Viral; Carcinoma; Epidermal Growth Factor; Epigenesis, Genetic; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Humans; Immunotherapy, Adoptive; Molecular Targeted Therapy; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Viral Matrix Proteins

To explore the effect of quadruple fluid aerosol inhalation on oral mucositis induced by radiotherapy in nasopharyngeal carcinoma (NPC) and its mechanism.. One hundred and twenty NPC patients were divided into a treatment group (n=60) and a control group (n=60). Radiation therapy was administered conventionally over 7 weeks with 2.0 Gy daily fractions for 5 days each week at the 70 Gy. Patients in the control group accepted oral care conventionally, and patients in the treatment group used quadruple fluids aerosol inhalation before the radiotherapy. The saliva volume was measured by a nurse twice a week and the concentration of epidemal growth factor (EGF) was examined by ELISA.. The number of patients with mucositis in the treatment group was significantly fewer than that in the control group when the total doses were up to 30 Gy and 60 Gy (P<0.05). The concentration of EGF in the control group significantly decreased when total doses were up to 30 Gy and 60 Gy (P<0.01). The concentration of EGF in the treatment group was not significantly different compared with pre-radiotherapy when the total doses were up to 30 Gy, but when total doses were up to 60 Gy, the concentration of EGF in the treatment group obviously decreased compared with that of pre-radiotherapy.. The quadruple fluid aerosol inhalation is effective to reduce oral mucositis resulted from radiotherapy in NPC patients, which is related to the delay of the EGF decrease in the saliva.

Topics: Administration, Inhalation; Adult; Aerosols; Aged; Drug Combinations; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Female; Gentamicins; Humans; Male; Menthol; Middle Aged; Nasopharyngeal Neoplasms; Procaine; Radiotherapy; Saliva; Stomatitis; Treatment Outcome; Vitamin B 12; Vitamins

Nasopharyngeal carcinoma (NPC) is a unique head and neck malignancy with highly metastatic cell-biological characteristics, for which latent EBV-infection is responsible. Our earlier studies showed that EGF-stimulated Ca

Topics: Animals; Calcium Signaling; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epidermal Growth Factor; Epithelial-Mesenchymal Transition; Epstein-Barr Virus Infections; Female; Humans; Mice; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Neoplasm Metastasis; Neoplasm Proteins; Neoplasm Transplantation; Stromal Interaction Molecule 1; Up-Regulation; Zebrafish

Store-operated Ca(2+) entry (SOCE) mediates Ca(2+) responses evoked by extracellular signaling molecules to promote increases in cytosolic Ca(2+), thereby triggering downstream signal transduction. Here we demonstrated that either the pharmacological blockage of Ca(2+) influx through SOCE or the knockdown of Orai1, a key molecule of SOCE, suppressed the epidermal growth factor-induced migration by disturbing Ca(2+) signaling in nasopharyngeal carcinoma (NPC) cell. Furthermore, Orai1 depletion led to a delayed cell attachment to the extracellular matrix surface in vitro and eliminated the extravasation of microinjected cells from vasculature in a zebrafish hematogenous metastasis model. Our findings thus indicate that SOCE acts as a predominant Ca(2+) signaling involved in NPC cell metastasis, and may serve as a candidate target for anti-metastasis therapy in NPC.

Topics: Animals; Animals, Genetically Modified; Calcium; Calcium Channel Blockers; Calcium Signaling; Carcinoma; Cell Line, Tumor; Cell Movement; Epidermal Growth Factor; Humans; Intracellular Calcium-Sensing Proteins; Ion Transport; Membrane Proteins; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Signal Transduction; Zebrafish

Dysregulation of E-cadherin and β-catenin function in cell-cell adhesion is common in nasopharyngeal carcinoma (NPC) and correlates with metastatic disease. In this study, we examined the role of EGF-activated phosphatidylinositol 3-kinase (PI3K)-Akt signaling in E-cadherin and β-catenin regulation. We found that reduced membranous E-cadherin and β-catenin expression was positively correlated with Akt phosphorylation in NPC tissues. EGF treatment disrupted cell-cell adhesion and resulted in mesenchymal morphological features in NPC cell lines (TW01, TW04, and TW06). Western blot analysis showed that the E-cadherin protein level was partially reduced in TW04 cells only and the β-catenin levels were not considerably affected upon EGF treatment. In contrast, quantitative real-time RT-PCR showed that the E-cadherin, but not β-catenin, mRNA levels were markedly reduced by EGF in all cell lines. Immunofluorescent staining revealed that E-cadherin and β-catenin appeared to be markedly reduced on the cell surface and more localized in the cytoplasm. Inhibition of PI3K by LY294002 did not abolish the EGF-induced downregulation of E-cadherin protein or mRNA in TW04 cells but moderately increased the β-catenin protein level in TW01 cells and mRNA level in TW06 cells. However, LY294002 substantially restored or increased cell surface E-cadherin and β-catenin in all EGF-treated cell lines, in concordance with the inhibition of cell morphological changes. Moreover, LY294002 significantly blocked EGF-driven cell invasion, correlating with the elevation of membranous E-cadherin and β-catenin levels. In conclusion, EGF-induced epithelial-to-mesenchymal transition may not be only dependent on downregulation of E-cadherin protein/mRNA but also on mislocalization of E-cadherin and β-catenin. The mechanisms involved may be related, at least in part, to the PI3K-Akt pathway.

Topics: Base Sequence; beta Catenin; Blotting, Western; Cadherins; Cell Line, Tumor; DNA Primers; Enzyme Activation; Epidermal Growth Factor; Humans; Immunohistochemistry; Microscopy, Fluorescence; Nasopharyngeal Neoplasms; Neoplasm Invasiveness; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Real-Time Polymerase Chain Reaction; Signal Transduction

Nasopharyngeal cancer (NPC) is an Epstein-Barr virus (EBV)-associated head and neck cancer prevalent in Asia. Although with reasons not fully understood, the intrinsic invasiveness of NPC is believed to be EBV-linked. Recently, EBV was found to induce STAT3 activation. Constitutive STAT3 activation correlated with advanced clinical staging in NPC. We hypothesized that STAT3 activation by EBV directly contributes to the intrinsic invasiveness of NPC cells. Phospho-STAT3-Tyr705 was detected in high percentage of NPC tumors (7/10 cases). Using a paired NPC cell line model, HONE-1 and the EBV-infected counterpart, HONE-1-EBV, we found that HONE-1-EBV expressed a higher level of phospho-STAT3-Tyr705 and was approximately 11-fold more invasive than HONE-1. In HONE-1-EBV, STAT3 siRNA targeting inhibited both spontaneous and serum-induced invasion, as well as cell growth. Conversely, activation of STAT3 (by expressing an activated STAT3 mutant, namely STAT3C) in the parental HONE-1, mimicking EBV-induced STAT3 activation, significantly enhanced its invasiveness and proliferation, which was accompanied by increased expression of markers of mesenchymal status, proliferation and anti-apoptosis. Our results demonstrated that EBV-induced STAT3 activation is responsible for NPC cell proliferation and invasion. This was further confirmed by a small molecule inhibitor of JAK/STAT3, JSI-124. JSI-124 inhibited STAT3 activation in HONE-1-EBV, with subsequent growth inhibition, induction of PARP cleavage, abrogation of anchorage-independent growth and invasion. We found that EBV-independent activation of STAT3 by a growth factor, EGF, also contributed to NPC invasion. In conclusion, EBV-induced STAT3 activation directly contributes to the intrinsic invasiveness of NPC cells and STAT3 targeting may be beneficial in treating aggressive NPC.

Topics: Blotting, Western; Cell Adhesion; Cell Movement; Cell Proliferation; Collagen; Colony-Forming Units Assay; Drug Combinations; Epidermal Growth Factor; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Humans; Laminin; Nasopharyngeal Neoplasms; Neoplasm Invasiveness; Proteoglycans; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; STAT3 Transcription Factor; Tumor Cells, Cultured

Human Cripto-1, a member of the EGF-CFC family, is indispensable for early embryonic development. Cripto-1 plays an important oncogenic role during tumorigenesis and is overexpressed in a wide range of epithelial carcinomas, yet little is known about Cripto-1 in nasopharyngeal carcinoma (NPC). The aim of this study was to analyze the roles of Cripto-1 in the progression and clinical characteristics in NPC clinical samples and cell lines.. The expression of Cripto-1 at mRNA level was detected by the reverse transcription-polymerase chain reaction (RT-PCR) and real time RT-PCR, and western blot was used to examine the protein expression. Cripto-1 expression and its clinical characteristics were investigated by performing immunohistochemical analysis on a total of 37 NPC clinical tissue samples. Lentiviral vectors were constructed to get an efficient expression of anti-Cripto-1 siRNA in CNE-2 and C666-1 cells, with invalid RNAi sequence as control. After the inhibition of the endogenous Cripto-1, the growth, cell cycle and invasion of cells were detected by MTT, FACS and Boyden chamber assay respectively. Moreover, in vivo, the proliferation of the tumor cells was evaluated in xenotransplant nude mice model with whole-body visualizing instrument.. The results of real-time RT-PCR and western blot showed that the expression level of Cripto-1 was markedly higher in NPC cell lines than that in the immortalized nasopharyngeal epithelial cell at both mRNA and protein levels. RT-PCR of 17 NPC tissues showed a high expression rate in 76.5% (13/17) cases. In an immunohistochemical study, Cripto-1 was found to express in 54.1% (20/37) cases of NPC. In addition, Cripto-1 overexpression was significantly associated with N classification (p = 0.034), distant metastasis (p = 0.036), and clinical stage (p = 0.007). Inhibition of endogenous Cripto-1 by lentivirus-mediated RNAi silencing technique suppressed NPC cell growth and invasion in vitro. In vivo, the average weight (p = 0.026) and volume (p = 0.044) of tumor in CNE-2/GFP+/Cripto-1- xenotransplant mice group were significantly lower than those in the control group. The Ki67 index was obviously lower in Cripto-1 RNAi treated tumors (p < 0.01).. Data of this study suggest that Cripto-1 overexpression is connected with the tumorigenesis and progression of NPC, lentivector-mediated RNAi might be feasible for the inhibition of the growth and invasion of NPC.

Topics: Adult; Aged; Aged, 80 and over; Animals; Carcinoma; Cell Line, Tumor; Epidermal Growth Factor; Female; Gene Expression Regulation, Neoplastic; GPI-Linked Proteins; Humans; Intercellular Signaling Peptides and Proteins; Male; Membrane Glycoproteins; Mice; Middle Aged; Nasopharyngeal Neoplasms; Neoplasm Proteins; Neoplasm Transplantation; Neoplastic Processes

Numerous candidate genes have been proposed as susceptibility factors for the development of nasopharyngeal carcinoma (NPC). Epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) interaction plays a pivotal role in cell proliferation, differentiation, and tumourigenesis of epithelial tissues. To our knowledge, however, no study has examined the relationship between the EGF/EGFR and NPC. The aim of this study is to investigate the potential association between single-nucleotide polymorphisms of EGF +61 G/A and EGFR +2073 A/T and NPC. A total of 173 patients with NPC and 206 age- and sex-matched controls were the participants. Genotypes were determined using a polymerase chain reaction-restriction fragment length polymorphism strategy and DNA sequencing. There were no significant differences in the genotype and allele frequencies of EGF +61 G/A and EGFR +2073 A/T polymorphisms between the group of patients with NPC and the control group in a Chinese population (for EGF +61 G/A: OR=1.29, 95% CI: 0.95-1.74; for EGFR +2073 A/T: OR=0.91, 95% CI: 0.67-1.23). Further studies are still needed to explore the complicated interaction between environmental factors and EGF +61 G/A and EGFR +2073 A/T polymorphisms in the risk of NPC, particularly in ethnically different populations.

Topics: Adult; Alleles; Base Sequence; Carcinoma, Squamous Cell; Case-Control Studies; DNA Primers; Epidermal Growth Factor; ErbB Receptors; Female; Gene Frequency; Humans; Male; Middle Aged; Nasopharyngeal Neoplasms; Polymorphism, Genetic

To investigate the inter-relationship of the expressions of cyclooxygenases-2 (COX-2), vascular endothelial growth factors (VEGF), and epidermal growth factor receptor (EGFR) in nasopharyngeal cancer (NPC) cells, and their clinical significance in association with the extent of disease at diagnosis.. Prospective.. Expressions of COX-2, VEGF, and EGFR protein were detected using immunohistochemistry in 111 patients with pathologically confirmed stage II to IV nasopharyngeal carcinoma. The correlation between the expressions of the three tumor markers and the stages of disease at diagnosis were investigated.. COX-2, VEGF, and EGFR were over-expressed in 76.6, 66.7, and 73.9% of NPC cells, respectively. The staining patterns was cytoplasmic for VEGF, membranous for EGFR, and both cytoplasmic and membranous for COX-2 in tumor cells. Linear associations were observed between the intensity of the expressions of COX-2 vs. VEGF, COX-2 vs. EGFR, or VEGF vs. EGFR. Furthermore, the intensity of the expressions of all three markers was significantly associated with the extent of the tumor measured by the Tumor, Node, Metastasis classification and staging grouping of the American Joint Committee on Cancer/International Union Against Cancer staging system.. COX-2, VEGF, and EGFR expressions in NPC cells were interrelated, and the intensity of the expressions of all three markers were significantly associated with the stage of the disease at diagnosis. Further investigation is needed to determine the clinical applications of COX-2, VEGF, and EGFR in predicting the long-term outcome of NPC after definitive therapy.

Topics: Biomarkers, Tumor; Biopsy; Cyclooxygenase 2; Diagnosis, Differential; Epidermal Growth Factor; Female; Follow-Up Studies; Humans; Immunohistochemistry; Magnetic Resonance Imaging; Male; Middle Aged; Nasopharyngeal Neoplasms; Neoplasm Staging; Prognosis; Retrospective Studies; Severity of Illness Index; Tomography, X-Ray Computed; Vascular Endothelial Growth Factor A

The epidermal growth factor (EGF)-like domain is involved in receptor-ligand interactions, extracellular matrix formation, cell adhesion and chemotaxis. Nasopharyngeal carcinoma associated gene 6 (NGX6) is a novel EGF-like domain-containing gene located at the high frequent loss of heterozygosity (LOH) region 9p21-22 associated with nasopharyngeal carcinoma (NPC). It is down-regulated in NPC and its over-expression can delay the cell cycle G(0)-G(1) progression in NPC cells. In the present study, in situ hybridization analysis, using NPC tissue microarrays, showed that loss of NGX6 expression was associated with NPC lymph node metastasis. The Tet-on gene expression system and cDNA array techniques were used to profile the potential targets of NGX6. We found that NGX6 can influence the expression of some cell adhesion molecules in NPC cells. NGX6 can associate with ezrin, a linkage between the cell membrane and cytoskeleton. The NGX6 protein was expressed on the cell surface as a glycoprotein. Ectopic induction of NGX6 can impair NPC cell migration and invasive ability as well as improve cell adhesion and gap junctional intercellular communication, and can suppress tumor formation in vivo. The data revealed that NGX6 plays a role in cell adhesion modulation in NPC cells.

Topics: Animals; Carcinoma; Cell Adhesion; Cell Adhesion Molecules; Cell Cycle; Cell Membrane; Cell Movement; Chlorocebus aethiops; COS Cells; Cytoskeletal Proteins; Cytoskeleton; Epidermal Growth Factor; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Membrane Proteins; Nasopharyngeal Neoplasms; Phosphoproteins; Tumor Cells, Cultured; Tumor Suppressor Proteins

To elucidate the regulation of the phosphorylation of epidermal growth factor receptor (EGFR) by the EB virus encoded latent membrane protein 1 (LMP1) in nasopharyngeal carcinoma cell line.. The levels of EGFR expression and phosphorylation in pTet-on LMP1 HNE2 cell, a nasopharyngeal carcinoma (NPC) cell line, in the dynamic expression of LMP1 induced by different concentrations of doxycycline (Dox) were observed. The EGFR dominant negative mutant and LMP1 antisense expression plasmid were transiently transfected into pTet-on LMP1 HNE2 cells by lipofectamine, and the changes in EGFR phosphorylation were observed by immunocoprecitation and Western blot. The changes in EGFR phosphorylation were observed after EGF treatment.. In pTet-on LMP1 HNE2 cells, Dox-induced LMP1 upregulated EGFR expression and phosphorylation in a dose-dependent manner. After EGFR dominant negative mutant was transfected into pTet-on LMP1 HNE2 cells, the increase of EGFR phosphorylation was inhibited completely. When LMP1 antisense expression plasmid was transfected into pTet-on LMP1 HNE2 cells, the levels of EGFR phosphorylation were also inhibited significantly. Meanwhile, after EGF had been added into pTet-on LMP1 HNE2 cells, increase of EGFR phosphorylation was induced, but it was completely blocked by EGFR dominant negative mutant and the introduction of LMP1 antisense.. EB virus encoded LMP1 not only induces the dose-dependent expression of EGFR, but also the dose-dependent phosphorylation of EGFR. The phosporylation of EGFR may play a vital role in the development of nasopharyngeal carcinoma.

Topics: Blotting, Western; Epidermal Growth Factor; ErbB Receptors; Herpesvirus 4, Human; Humans; Nasopharyngeal Neoplasms; Phosphorylation; Tumor Cells, Cultured; Viral Matrix Proteins

To examine the IP3 amounts, invasive potentials, and response to LN and EGF of the substrains with high or low metastatic ability from two kinds of carcinoma cell lines (MFC and CNZ-2Z).. Radio-ligand binding assay and matrigel invasive assay.. The low metastatic substrains from the two lines had higher amounts of IP3 and stronger response to EGF than their responsive high ones, but high metastatic substrains from CNZ-2Z had stronger response to LN than the low ones while MFC substrains had not significant difference in response to LN.. There is internal difference in signal transduction between the high and low metastatic cancer cells, and it is significant to study the difference in detail.

Topics: Animals; Epidermal Growth Factor; Humans; Inositol 1,4,5-Trisphosphate; Laminin; Mice; Nasopharyngeal Neoplasms; Neoplasm Metastasis; Signal Transduction; Stomach Neoplasms; Tumor Cells, Cultured

EBV-LMP gene transfection, radioimmuno-binding assay (RIA) and colorimetric cell viability assay (MTT) were used to investigate the effect and relationship of EBV-LMP and EGF autocrine on the growth and proliferation of well differentiated nasopharyngeal carcinoma cell line (CNE1). Autocrine EGF was detected in the supernant of CNE1 cells and CNE1 cells could grow in serum-free medium, implicating that the cell proliferation-promoting effect of EGF autocrine was present in the CNE1 cell line. Nevertheless the positive signals of LMP gene polymerase chain reaction and LMP Mab immunohistochemical staining in pCMV alpha-LMP DNA transfected cells, confirmed the successful transfection of LMP gene. EGF autocrine amount and the proliferation of CNE1 cells in serum-free medium were more obvious in post-transfected cells than those in untransfected cells. The present study represents the first report on promotion of EGF autocrine in EBV-LMP-transfected cells, thus promoting cell proliferation.

Topics: Autocrine Communication; Cell Transformation, Viral; Epidermal Growth Factor; Herpesvirus 4, Human; Humans; Nasopharyngeal Neoplasms; Oncogene Proteins, Viral; Tumor Cells, Cultured; Viral Matrix Proteins

Epidermal growth factor (EGF), amylase and haptocorrin are molecules produced in the salivary glands. The aim of the present study was to determine immunohistochemical and quantitative alterations in EGF as compared with haptocorrin and amylase following radiotherapy for oral cancer. Changes in the salivary secretion of EGF are of interest because of the importance of EGF in mucosal regeneration. Immunohistochemical studies on normal tissue from parotid and submandibular glands have demonstrated EGF in the serous acini with a tendency to single cell expression in the parotid gland. Amylase has been found in the serous acini of both the submandibular and parotid glands. Haptocorrin was localized in the duct system of both glands. In the submandibular glands with radiotherapy induced sialoadenitis only very few acini with weak or no staining for EGF and amylase were demonstrated, while no changes were observed in the staining for haptocorrin. Analysis on stimulated whole saliva samples collected from 20 healthy individuals and from 20 patients prior to, and 1, 2 and 3 weeks following radiotherapy showed significant reduction in salivary contents of EGF and amylase after treatment as expressed per g protein (p < 0.0002). The salivary content of haptocorrin increased significantly after treatment (p < 0.002). These alterations may be explained by the different cellular sites of the molecules studied, the serous acini being more sensitive to ionising radiation than the duct system. The concentration of EGF in saliva before treatment was significantly higher in patients than in the control group (p < 0.02), which may indicate that the tumors induce increased secretion of salivary EGF, or alternatively that the oral tumors contribute with EGF to the saliva. In conclusion we have demonstrated a reduction in the mitogenic peptide EGF both immunohistochemically and quantitatively following irradiation for oral cancer, results which may contribute to the understanding of the clinical signs of mucositis.

Topics: Aged; Amylases; Carcinoma, Squamous Cell; Epidermal Growth Factor; Female; Humans; Immunohistochemistry; Male; Middle Aged; Mouth; Mouth Neoplasms; Nasopharyngeal Neoplasms; Nasopharynx; Salivary Glands

CG-1 human nasopharyngeal carcinoma cells in monolayer culture formed both cohesive, epithelial-like colonies and scattered, fibroblastic-like colonies in mixed proportions. In the presence of exogenously added bFGF (4 ng/ml), about 85% of the colonies formed were fibroblastic-like. CG-1 cells were capable of synthesizing and releasing bFGF, and, when compared by the immunological method, cells in fibroblastic-lke colonies were found to contain higher levels of endogenous bFGF than cells in the epithelial-like colonies. Furthermore, cells in the peripheral region of the epithelial-like colonies, which were fibroblastic-like in morphology, also appeared to contain higher levels of endogenous bFGF. In addition, in the presence of suramin, neutralizing antibody to bFGF, or neutralizing antibodies to bFGF and EGF, the number of cohesive colonies formed was greatly increased. Moreover, addition of the 2 M NaCl-eluted heparin-Sepharose fraction of the CG-1 cell-coditioned medium promoted the formation of dispersed colony in a dose-dependent manner. The results suggest that bFGF can regulate CG-1 cell phenotype in an autocrine manner.

Topics: Antibodies; Carcinoma; Epidermal Growth Factor; Epithelial Cells; Fibroblast Growth Factor 2; Fibroblasts; Humans; Mesoderm; Nasopharyngeal Neoplasms; Neoplastic Stem Cells; Suramin; Tumor Cells, Cultured

The growth of a recently established human nasopharyngeal carcinoma cell line, CG-1, and 5 randomly selected, single-cell-derived sub-lines in serum-free medium and in fetal-bovine-serum(FBS)-containing medium was investigated. In basal medium supplemented with insulin, transferrin, fibronectin and high-density lipoprotein, cell growth was moderately stimulated by aFGF and EGF in a dose-dependent manner. In contrast, in medium containing as little as 0.5% FBS, most of the stimulatory effect of the aforementioned growth factors observed was masked. Western blotting analysis of the cell lysates and conditioned media showed that CG-1 and sub-line cells were all capable of synthesizing and releasing aFGF- and EGF-immunoreactive proteins. The amounts of these 2 growth factors synthesized and released appeared to vary among the parental cell line and sub-line cells. Moreover, the rate of basal proliferation of these cells appeared to be positively correlated with the amounts of aFGF- and EGF-immunoreactive proteins produced. Addition of the neutralizing antibodies to aFGF and EGF exerted a dose-dependent suppression on cell growth in medium containing 0.5% FBS. The results suggest a role of aFGF and EGF in autocrine growth stimulation of CG-1 and sub-line cells, and may explain the moderate response of these cells to exogenously added aFGF and EGF.

Topics: Carcinoma; Cell Division; Culture Media, Serum-Free; Dose-Response Relationship, Drug; Epidermal Growth Factor; Fibroblast Growth Factor 1; Humans; Nasopharyngeal Neoplasms; Tumor Cells, Cultured