Growth Of C6 Cells Research Articles

Eslicarbazepine induces apoptosis and cell cycle arrest in C6 glioma cells in vitro and suppresses tumor growth in an intracranial rat model

BackgroundGlioblastoma multiforme (GBM) is the most malignant brain tumor, with a poor prognosis and life expectancy of 14–16 months after diagnosis. The standard treatment for GBM consists of surgery, radiotherapy, and chemotherapy with temozolomide. Most patients become resistant to treatment after some time, and the tumor recurs. Therefore, there is a need for new drugs to manage GBM. Eslicarbazepine (ESL) is a well-known antiepileptic drug belonging to the dibenzazepine group with anticancer potentials. In this study, for the first time, we evaluated the potential effects of ESL on C6 cell growth, both in vitro and in vivo, and examined its molecular effects.MethodsTo determine the effect of ESL on the c6 cell line, cell viability, proliferation, and migration were evaluated by MTT assay, colony formation, and wound healing assay. Also, apoptosis and cell cycle were examined by flow cytometry, qRT-PCR, and western blotting. In addition, an intracranial model in Wistar rats was used to investigate the effect of ESL in vivo, and the tumor size was measured using both Caliper and MRI.ResultsThe obtained results are extremely consistent and highly encouraging. C6 cell viability, proliferation, and migration were significantly suppressed in ESL-treated C6 cells (p < 0.001), as determined by cell-based assays. ESL treatment led to significant enhancement of apoptosis (p < 0.01), as determined by flow cytometry, and upregulation of genes involved in cell apoptosis, such as the Bax/Bcl2 ratio at RNA (p < 0.05) and protein levels (5.37-fold). Flow cytometric analysis of ESL-treated cells revealed G2/M phase cell cycle arrest. ESL-treated cells demonstrated 2.49-fold upregulation of p21 alongside, 0.22-fold downregulation of cyclin B1, and 0.34-fold downregulation of cyclin-dependent kinase-1 at the protein level. Administration of ESL (30 mg/kg) to male rats bearing C6 intracranial tumors also suppressed the tumor volume and weight (p < 0.01).ConclusionsBased on these novel findings, ESL has the potential for further experimental and clinical studies in glioblastoma.Graphical abstract

Stimuli-Responsive Micelles with Detachable Poly(2-ethyl-2-oxazoline) Shell Based on Amphiphilic Polyurethane for Improved Intracellular Delivery of Doxorubicin.

Polyurethanes (PUs) have various biomedical applications including controlled drug delivery. However, the incompletely release of drug at tumor sites limits the efficiency of these drug loaded polyurethane micelles. Here we report a novel polymer poly(2-ethyl-2-oxazoline)-SS-polyurethane-SS-poly(2-ethyl-2-oxazoline) triblock polyurethane (PEtOz-PU(PTMCSS)-PEtOz). The hydrophilic pH-responsive poly(2-ethyl-2-oxazoline) was used as an end-block to introduce pH responsiveness, and the hydrophobic PU middle-block was easily synthesized by the reaction of poly (trimethylene carbonate) diol containing disulfide bonds (PTMC-SS-PTMC diol) and bis (2-isocyanatoethyl) disulfide (CDI). PEtOz-PU(PTMCSS)-PEtOz could self-assemble to form micelles (176 nm). The drug release profile of PEtOz-PU(PTMCSS)-PEtOz micelles loaded with Doxorubicin (DOX) was studied in the presence of acetate buffer (10 mM, pH 5.0) and 10 mM dithiothreitol (DTT). The results showed that under this environment, DOX-loaded polyurethane micelles could release DOX faster and more thoroughly, about 97% of the DOX was released from the DOX-loaded PEtOz-PU(PTMCSS)-PEtOz micelle. In addition, fluorescent microscopy and cell viability assays validated that the DOX-loaded polyurethane micelle strongly inhibits the growth of C6 cells, suggesting their potential as a new nanomedicine against cancer.

Natural product-derived icaritin exerts anti-glioblastoma effects by positively modulating estrogen receptor β.

Glioblastoma is the most common malignancy of the central nervous system, and patients typically have a poor prognosis. Previous studies indicate a gender bias in the development of glioblastoma; women are at a lower risk compared with men, suggesting that estrogen may confer protective effects. Icaritin, a prenylflavonoid derivative from a Chinese herb of the Epimedium genus, selectively regulates the estrogen receptor (ER) and possesses anti-cancer properties. The aim of the present study was to investigate the protective effects of icaritin on glioblastoma and its underlying mechanisms, with a particular focus on its association with the ER. The results demonstrated that icaritin inhibited the growth of C6 and U87-MG glioblastoma cells in a dose- and time-dependent manner. At a concentration of 12.5 µM, icaritin induced apoptosis, which was characterized by the increased expression of the cleaved forms of caspases 3, 7, 8 and 9 and poly (ADP-ribose) polymerase, downregulation of BCL2 apoptosis regulator and upregulation of BCL2-associated X, apoptosis regulator expression. Additionally, icaritin inhibited the migration of C6 and U87-MG cells. The protein expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 were also downregulated following icaritin treatment. Furthermore, icaritin treatment increased the expression of estrogen receptor (ER)β and the phosphatase and tensin (PTEN) homolog oncoprotein, thus reducing the expression of downstream targets of PTEN; protein kinase B (Akt) and phosphorylated Akt. Subsequent experiments demonstrated that icaritin cooperates with 17β-estradiol to inhibit the growth of glioblastoma cells, and the inhibition of ERβ with the ERβ-specific antagonist ICI 182,780, attenuated the anti-glioblastoma effects of icaritin. In conclusion, the results of the present study demonstrate that the anti-glioblastoma effects of icaritin may be mediated by its modulation of ERβ.

XCHD Inhibits C6 Cell Growth Primarily via the p53/Caspase Pathway.

The effects of XCHD on the proliferation of C6 cells and on factors associated with the microRNA-34a (miR-34a)/p53/caspase-3 signaling pathway in vitro were investigated. Methods. XCHD was purchased too much to complete the study. CCK-8 assay was used to measure the XCHD concentration, and qPCR was used to quantify miR-34a expression at the mRNA level. Apoptosis was assessed using TUNEL. Western blots were used to determine the p53, caspase-3, caspase-8, and Bcl-2 expression levels. Results. The optimal XCHD concentration and time effect for C6 cells were observed after 36 h of exposure to a concentration of 100 µg/ml XCHD. miR-34a expression increased 8 and 12 h after the addition of XCHD. The presence of XCHD decreased Bcl-2 expression but increased p53, cleaved caspase-3, Bax, and caspase-8 expression. When p53 was inhibited, miR-34a expression was unaffected by the addition of XCHD, Bcl-2 expression was low, and cleaved caspase-3, Bax, and caspase-8 expression increased. The inhibition of p53 promoted C6 cell growth when compared with C6 cells exposed to XCHD and with no inhibition of p53. Conclusions. XCHD inhibits C6 cell growth which was influenced by the p53/caspase pathway.

Synergistic Anticancer Effects of Formononetin and Temozolomide on Glioma C6 Cells

Temozolomide (TMZ) is currently the first-line drug used for clinical postoperative or non-surgical chemotherapy for glioma, but acquired and intrinsic resistance to TMZ limits its application. The anti-proliferative effect of formononetin on human glioma cells had been confirmed. To improve therapeutic effects of TMZ, we studied the effect of formononetin in combination with TMZ on C6 glioma cells. The anti-proliferative effect of C6 cells was tested by 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. The synergy was evaluated by Chou-Talalay method. Morphological changes were observed by hematoxylin-eosin (HE) staining. The effect of formononetin in combination with TMZ on apoptosis of C6 cells was investigated by flow cytometry. The effect of formononetin in combination with TMZ on migration of cells was investigated by wound healing assay and transwell assay. The expression of proteins related to apoptosis and migration were detected by Western blot. These results showed that formononetin or TMZ alone could inhibit the growth of C6 cells in dose-dependent manner and formononetin in combination with TMZ had synergy effect on C6 cells. Further changes in cell morphology could be observed in drug combination by HE staining. Drug combination enhanced the expression of Bax, Cleaved Caspase-3, Cleaved Caspase-9, decreased the expression of Bcl-2, and promoted tumor cells apoptosis. In addition, the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) were down-regulated via drug combination which resulted into inhibiting migration of C6 cell. In conclusion, formononetin in combination with TMZ can play a synergistic role in anti-C6 cells, the mechanisms of synergy depended on multiple pathways.

Temozolomide inhibits cellular growth and motility via targeting ERK signaling in glioma C6 cells.

Temozolomide (TMZ) is an alkylating agent used for the treatment of aggressive forms of brain tumor based on its antitumor actions. However, the exact effect on cancer and the underlying anticancer molecular mechanism of TMZ remain to be elucidated. In the present study, the effects of TMZ on the growth and motility of glioma C6 cells were investigated. MTT and Transwell assays were used to detect cellular growth and motility. The results showed that TMZ inhibited the proliferation, migration and invasion of the glioma C6 cells invitro, western blot analysis determined that the phosphorylation of extracellular signal‑regulated protein kinase (ERK)1/2 was decreased in the TMZ‑treated cells, compared with the untreated control cells. The ERK1/2 specific inhibitor, U0126, augmented the inhibitory effects of TMZ on the proliferation, migration and invasion of the glioma C6 cells, and the mitogen‑activated protein kinase kinase/ERK pathway activator, curcumin, attenuated the inhibitory effects of TMZ on the proliferation and motility of the glioma C6 cells. Additionally, the western blotting in the present study demonstrated that TMZ and U0126 decreased the expression of vascular endothelial growth factor-C (VEGF-C), and the expression level was restored by curcumin, suggesting that VEGF‑C may be the downstream effector of ERK1/2. Furthermore, the overexpression of VEGF‑C enhanced the growth, migration and invasion of the TMZ-treated cells. These results suggested that TMZ suppressed glioma C6 cell development, at least in part, and downregulated the expression of VEGF‑C by inhibiting the ERK signaling pathway. The results of the present study provides the foundation for a combinational therapeutic strategy to improve the efficacy of TMZ.

The flavonoid apigenin from Croton betulaster Mull inhibits proliferation, induces differentiation and regulates the inflammatory profile of glioma cells.

This study aimed to investigate the antitumor and immunomodulatory properties of the flavonoid apigenin (5,7,4'-trihydroxyflavone), which was extracted from Croton betulaster Mull, in glioma cell culture using the high-proliferative rat C6 glioma cell line as a model. Apigenin was found to have the ability to reduce the viability and proliferation of C6 cells in a time-dependent and dose-dependent manner, with an IC50 of 22.8 µmol/l, 40 times lower than that of temozolomide (1000 µmol/l), after 72 h of apigenin treatment. Even after C6 cells were treated with apigenin for 48 h, high proportions of C6 cells entered apoptosis (39.56%) and autophagy (22%) as shown by flow cytometry using annexin V/propidium iodide and acridine orange staining, respectively. In addition, the flavonoid apigenin induced cell accumulation in the G0/G1 phase of the cell cycle and inhibited glioma cell migration efficiently. Moreover, apigenin induced astroglial differentiation and morphological changes in C6 cells, characterized by increased expression of glial fibrillary acidic protein and decreased expression of nestin protein, a typical marker of neuronal precursors. The immunomodulating effects of apigenin were also characterized by a change in the inflammatory profile as evidenced by a significant decrease in interleukin-10 and tumor necrosis factor production and increased nitric oxide levels. Because apigenin can induce differentiation, apoptosis, and autophagy, can alter the profile of cytokines involved in regulating the immune response, and can reduce the survival, growth, proliferation, and migration of C6 cells, this flavonoid may be considered a potential antitumor drug for the adjuvant treatment of malignant gliomas.

Inhibiting the growth of tumor cells by ruthenium(II) complexes [Ru(phen)2L] (L = o-TFMPIP and p-CPIP) through DNA-binding

[Ru(phen)2o-TFPIP]2+ (1) and [Ru(phen)2p-CPIP]2+ (2) have been synthesized and demonstrated to inhibit the growth of tumor cells. The inhibitory activities (IC50) of 1 against the growth of C6, MDA-MB-231 and HepG2 cells were about 24.5, 36.7, and 36.1 μM, respectively. Studies show that both complexes bind to CT-DNA, explained by using density functional theory calculations. The LogP calculated for 1 and 2 are −0.4859 and −1.279, respectively. These complexes, especially 1, can be used as promising inhibitors in chemotherapy, and their DNA-binding behaviors play a key role.

Ozone Dosage Effect on C6 Cell Growth: in Vitro and in Vivo Tests.

C6 rat glioma cells are one of the most aggressive carcinogenic tumors, due to its high mortality rate in human beings and animals. The current treatment for this illness includes surgery, radio and chemotherapy, showing relapse in patients treated with those therapies. Since the ozone was found to be an effective bioreactive to inhibit growth of several carcinoma cells in vitro and in vivo. In this research, therapeutic peritoneum insufflation of ozone/oxygen dissolved in the physiological solution of NaCl 0.9% was dosed for fifteen days on different female mice groups in an advanced stage of C6 tumor (n=6). The first of them was the control group which had no treatment, the second group was dosage with oxygen every second day, the third group was dosed with ozone every second day, and finally the fourth group was dosed with ozone dissolved every fifth day. The size of the tumor was higher in both groups dosage by ozone, nevertheless tumor activity measured by microPET was 98% less in the fourth group compared with the control group. That result proves that ozone provokes an increase in the tumor volume even though the decrease of the cell activity. Those results were confirmed by the quantification of hydroperoxides, total cholesterol and total triglycerides.

Abstract 2528: Vardenafil is superior to idarubicin in treatment of glioma

Abstract Idarubicin (IDA) is a very potent second generation anthracycline that has shown high efficacy against glioma in vitro. However, this agent has failed to exert satisfactory therapeutic response in patients with brain tumors. Phosphodiesterase (PDE-5), an enzyme which hydrolyzes intracellular cyclic guanosine monophosphate (cGMP), is highly expressed in many brain tumor cell lines. We proposed that increasing cGMP in the brain by the potent PDE-5 inhibitor vardenafil (VAR) will augment the antitumor efficacy of IDA against glioma. We also proposed that VAR will induce apoptosis of glioma cells. Here we show for the first time that VAR (10 mg/kg) enhanced the antitumor activity of IDA (0.1mg/kg). VAR alone inhibited tumor growth and prolonged survival of glioma bearing rats. Histopathological examination revealed that tumors from glioma bearing rats treated with VAR alone or VAR+ IDA presented a significant reduction in mitotic index, indicating less proliferative tumors, as compared to those which received PBS (control) or IDA. Real time PCR results demonstrated that VAR inhibited mRNA expression of PDE-5 in the brains of animals treated with VAR, compared to control-treated group, implicating an important role for the inhibition of PDE-5 in the antitumor activity of VAR. Interestingly, the antitumor activity of VAR was similar to that of IDA. Moreover, animals receiving either VAR alone or VAR+IDA demonstrated statistically significant improvement in overall survival when compared with control (PBS) or IDA groups. In order to explain such an unexpected finding, we examined the antiproliferative, pro-apoptotic and anti-invasive activities VAR in vitro. MTT assay results showed that VAR significantly inhibited the growth of rat C6 glioma cells. Annexin V propidium iodide assay results revealed that the inhibition of C6 cell growth is mediated at least in part by inducing C6 apoptosis. Wound healing and soft agar colony formation assays showed that VAR inhibited the migration and anchorage-independent growth of C6 cells, respectively. This suggests that the enhanced antitumor effect of VAR could be due to a combined antiproliferative, pro-apoptotic and anti-invasive activities. Together, our data support anti-glioma properties of VAR, and provide a proof-of-concept basis for further development of VAR as potential adjuvant for treatment of brain tumors. Citation Format: Abdelkader E. Ashour, Abdulrahman Z. Alzahrani, Hala E. Abdel-Hamied, Khairy M. Zoheir, Sheikh F. Ahmed, Salem S. Al-Rejaie, Adel R. Abd-Allah. Vardenafil is superior to idarubicin in treatment of glioma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2528. doi:10.1158/1538-7445.AM2015-2528

Mechanical properties and permeability of porous chitosan–poly(p-dioxanone)/silk fibroin conduits used for peripheral nerve repair

Some poly(p-dioxanone) (PDO) homopolymers were first synthesized and the selected PDO was conjugated onto chitosan using a group-protecting method to produce chitosan–poly(p-dioxanone) (CH–PDO) copolymers with various PDO percentages changing from around 30 to 60wt%. The CH–PDO with the PDO content of around 42wt% was used to blend with prescribed amounts of silk fibroin (SF) to build porous single-lumen conduits that are intended to be used for long-gap peripheral nerve repair. Some genipin-crosslinked CH–PDO/SF conduits were endowed with an average porosity of around 60% in their porous wall, and with changed pore-sizes varying from around 10 to ca. 70μm using optimized processing conditions. After being degraded in a PBS medium containing a certain amount of lysozyme for various periods up to 8 weeks, some optimal CH–PDO/SF conduits were able to retain their compressive load and deformation recovery at around 59N/m and 73% in wet state, respectively. In addition, the achieved CH–PDO/SF conduits allowed the permeation of nutritional molecules with various molecular weights while showing a certain ability to prevent cells from infiltrating through the conduit wall. Cell culture confirmed that the optimized CH–PDO/SF conduits were able well supported the growth of rat glioma C6 cells. These results suggest that presently developed CH–PDO/SF conduits have promising potential for long-gap peripheral nerve repair.

Fatsioside A, a Rare Baccharane-Type Glycoside Inhibiting the Growth of Glioma Cells from the Fruits of Fatsia japonica

A novel baccharane-type triterpenoid glycoside named fatsioside A (1), together with ten oleanane glycosides, were isolated from the fruits of Fatsia japonica. The structure of fatsioside A was assigned as 3β,15α,18α-trihydroxy-18,19-secolupane-12,19-dione 3-O-β-D-glucopyranosyl-(1 → 2)-β-D-glucopyranoside by extensive NMR and HRESIMS analyses. F. japonica is the third baccharane glycoside-containing species reported to date in the plant kingdom, while fatsioside A represents the first baccharane glycoside found in the Araliaceae family. Fatsioside A inhibited the growth of rat glioma C6 cells and human glioma U251 cells with IC50 values of 33.48 ± 2.01 µM and 77.58 ± 6.19 µM, respectively. Further investigation indicated that fatsioside A induced apoptosis and necrosis in glioma cells, and arrested the cell cycle at the G0/G1 phase.

Ozone dosage effect on C6 cell growth, in vitro and in vivo tests: double bond index for characterization

Ozone dissolved in a saline solution applied as a medical therapy promoted an 85% decrement in C6 tumor activity.

Human umbilical cord mesenchymal stem cells inhibit C6 glioma growth via secretion of dickkopf-1 (DKK1)

Mesenchymal stem cells (MSCs) represent a potential therapeutic target for glioma. We determined the molecular mechanism of inhibitory effect of human umbilical cord-derived MSCs (hUC-MSCs) on the growth of C6 glioma cells. We demonstrated that hUC-MSCs inhibited C6 cell growth and modulated the cell cycle to G0/G1 phase. The expression of β-catenin and c-Myc was downregulated in C6 cells by conditioned media from hUC-MSCs, and the levels of secreted DKK1 were positively correlated with concentrations of hUCMSCs-CM. The inhibitory effect of hUC-MSCs on C6 cell proliferation was enhanced as the concentration of DKK1 in hUCMSCs-CM increased. When DKK1 was neutralized by anti-DKK1 antibody, the inhibitory effect of hUC-MSCs on C6 cells was attenuated. Furthermore, we found that conditioned media from hUC-MSCs transfection with siRNA targeting DKK1 mRNA or pEGFPN1-DKK1 plasmid lost or enhanced the abilities to regulate the Wnt signaling in C6 cells. Therefore, hUC-MSCs inhibited C6 glioma cell growth via secreting DKK1, an inhibitor of Wnt pathway, may represent a novel therapeutic strategy for malignant glioma.

Inhibitory effect of pseudolaric acid B on proliferation of rat glioma C6 cells

Objective To observe the inhibitory effects of pseudolaric acid B(PAB) on the proliferation of rat glioma C6 cells in vivo and in vitro.Methods MTT assay was used to examine the inhibitory effect of 0,0.1,0.2,0.4,0.8,1.6,8,and 16 μg/ml PAB on C6 cells;the distribution of cell cycle was determined by flow cytometry.C6 cells were injected subcutaneously to establish glioma-bearing rat model and the tumor volume was measured.The expression of proliferating cell nuclear antigen(PCNA) was detected immunohistochemically in the implanted glioma.Results In vitro results showed that PAB inhibited C6 cell growth in a dose-dependent manner,with the IC50 being 0.03 μg/ml.It also arrested the cells in G2/M phase.In vivo results showed that the tumor size of PAB group was decreased compared with the control group;moreover,PCNA expression was also decreased by PAB.Conclusion PAB can inhibit rat glioma C6 cell growth,which might be associated with cell arrest in G2/M phase and down-regulation of PCNA expression.

In vivo magnetic resonance imaging tracking of C6 glioma cells labeled with superparamagnetic iron oxide nanoparticles

The aim of the current study was to monitor the migration of superparamagnetic iron oxide nanoparticle (SPION)-labeled C6 cells, which were used to induce glioblastoma tumor growth in an animal model, over time using magnetic resonance imaging (MRI), with the goal of aiding in tumor prognosis and therapy. Two groups of male Wistar rats were used for the tumor induction model. In the first group (n=3), the tumors were induced via the injection of SPION-labeled C6 cells. In the second group (n=3), the tumors were induced via the injection of unlabeled C6 cells. Prussian Blue staining was performed to analyze the SPION distribution within the C6 cells in vitro. Tumor-inducing C6 cells were injected into the right frontal cortex, and subsequent tumor monitoring and SPION detection were performed using T2- and T2*-weighted MRI at a 2T field strength. In addition, cancerous tissue was histologically analyzed after performing the MRI studies. The in vitro qualitative evaluation demonstrated adequate distribution and satisfactory cell labeling of the SPIONs. At 14 or 21 days after C6 injection, a SPION-induced T2- and T2*-weighted MRI signal reduction was observed within the lesion located in the left frontal lobe on parasagittal topography. Moreover, histological staining of the tumor tissue with Prussian Blue revealed a broad distribution of SPIONs within the C6 cells. MRI analyses exhibit potential for monitoring the tumor growth of C6 cells efficiently labeled with SPIONs.

Inhibitive effect of C-21 steroidal glycosides of Cynanchum auriculatum on rat glioma cells in vitro

To evaluate the inhibitive effect of C-21 steroidal glycosides from the root of Cynanchum auriculatum (CGB) on rat glioma C6 cells. C6 cells were treated with CGB for 24, 48,72 h at concentration of 30, 60, 120 mg/L, respectively. MTT assay was used for evaluating cell viability; fluorescence-activated cell sorting analysis after Annexin V/propidium iodide staining or single propidium iodide staining was used to test cell apoptosis and cell cycle. CGB at 30, 60, 120 mg/L concentration-dependently decreased C6 cell viability (P<0.001). CGB at 60 and 120 mg/L induced C6 cell apoptosis and cell cycle arrest. The fraction of G0/G1 cells was increased (P<0.05) and that of S phase cells was decreased (P<0.01). CGB can inhibit the growth of rat glioma C6 cells, and induce apoptosis and G0/G1 cell cycle arrest.

Somatostatin receptor 1,2 and 5 activation leads to C6 glioma growth arrest in vitro and in vivo; analysis of the intracellular pathways involved

We report that somatostain receptor (SSTR) 1,2 and 5 activation by selective agonista, cause C6 cell growth arrest through PTPn-dependent dephosphorylation of ERK1/2 in vitro and after xenografting in nude mice. Individual SSTR agonist desplayed different efficacy and potecy showing partial synergism by combined treatment. Since most tumor cells express multiple SSTRs, the activtion of all the subtypes may grant a better control of cancer growth.

Naringenin, a flavanone alters the tumorigenic features of C6 glioma cells

The effect of naringenin on the growth of C6 glioma cells was investigated in the current study. Suppression of cell growth was assessed by MTT assay. Induction of cell cycle arrest was assessed by propidium iodide staining using flowcytometric analysis. Induction of DNA damage was assessed by agarose gel electrophoresis. The antiproliferative effect of NGEN was also assessed and confirmed by cyclooxygenase-2 (COX-2) activity assay. Gap junctional analysis was also done using dye transfer technique. The results showed that NGEN inhibited the growth of C6 cells in a dose dependent manner, compared to the control. At 20 and 30 μg concentrations, NGEN induced cell cycle arrest, induced DNA damage in C6 cells, reduced the activity of COX-2 and increased the gap junction intracellular communication (GJIC). Thus NGEN, a component of citrus fruit, suppresses the growth of C6 cells through the inhibition of proliferation.

Naringenin, a flavanone alters the tumorigenic features of C6 glioma cells

The effect of naringenin on the growth of C6 glioma cells was investigated in the current study. Suppression of cell growth was assessed by MTT assay. Induction of cell cycle arrest was assessed by propidium iodide staining using flowcytometric analysis. Induction of DNA damage was assessed by agarose gel electrophoresis. The antiproliferative effect of NGEN was also assessed and confirmed by cyclooxygenase-2 (COX-2) activity assay. Gap junctional analysis was also done using dye transfer technique. The results showed that NGEN inhibited the growth of C6 cells in a dose dependent manner, compared to the control. At 20 and 30 μg concentrations, NGEN induced cell cycle arrest, induced DNA damage in C6 cells, reduced the activity of COX-2 and increased the gap junction intracellular communication (GJIC). Thus NGEN, a component of citrus fruit, suppresses the growth of C6 cells through the inhibition of proliferation.