[Relationship between P65 and radiotherapy-induced oral squamous cell carcinoma cell line apoptosis].

OBJECTIVE: To investigate the effects of different X-ray doses on the expression of nuclear factor-kappaB (NF-kappaB) P65 in human oral squamous cell carcinoma cell (OSCC) line and the relationship between NF-kappaB P65 and radiation-induced OSCC cell line apoptosis. METHODS: The squamous cell carcinoma of Tca8113 cell was cultivated in the 37 degrees C, 5% CO2 incubator after recovery. The experiment samples were divided into six groups (control group, 2, 4, 6, 8, 10 Gy). After growing to logarithm period, Tca8113 cells were irradiated using above-mentioned X-ray doses. The immunocyteochemistry and Western blot were used to detect the expression of NF-kappaB P65 after irradiation in various times (1, 3, 6, 10, 24, 48 h). The apoptosis rates under different radiotherapy dose were detected by flow cytometer and TDT-mediated dUTP-biotin nick end labeling (TUNEL). RESULTS: Compared with the control group, cytoplasm expression of P65 under different X-ray doses had statistically significant differences (P < 0.05). While the cytoplasm P65 protein expression at different time were compared each other, the 3 h group demonstrated significant difference (P < 0.05). Apoptosis rates in various groups, compared with control group, had statistically significant differences (P < 0.05). While the groups at different time points were compared each other, the apoptosis rates of 3 h group had significant differences (P < 0.05). CONCLUSION: X-ray can activate the NF-kappaB P65 in oral squmaous cell carcinoma cell lines. The correlation between expressional quantity of P65 and radiotherapy induced apoptosis of oral squamous cell carcinoma cell lines possesses positive correlation. The activated and intranuclear P65 may have radiotherapy resistant effect.

[Experimental study on the expression of NF-κB-P65 protein and apoptosis of adenoid cystic carcinoma cells after radiation treatment].

PURPOSE: To detect the expression of p65, a subunit of NF-κB proteins, and apoptosis after adenoid cystic carcinoma cells(ACC-2) irradiated by high energy X-ray, and to investigate the interaction between them. METHODS: ACC-2 cells were cultured and then irradiated by high energy X-ray of different dose(2, 4, 6, 8,10Gy). At the next six time points(1, 3, 6, 10, 24, 48h), the expression of p65 protein in cytoplasm and nucleus was detected by immunocytochemistry and Western blotting. The apoptotic cells were counted by flow cytometry and then observed by TUNEL technique. The data of radiant intensity and apoptotic rate were statistically analyzed by Spearman method with SPSS11.5 software package. RESULTS: In ordinary condition, p65 protein seldom appeared in the nucleus, and mostly stained in the cytoplasm by immunocytochemistry. After irradiation, the protein was observed around the nuclear. Then it went through the nuclear membrane more and more as time going on, finally to the center of the nucleus. The quantity of p65 among the total protein changed gradually after radiation, rising at first, which got to a peak after about 6 to 10 hours, according to the results of Western blotting. At the same time point, p65 protein was found to have a higher expression with a higher dose of irradiation correspondingly. The proportion of apoptotic cells also varied from time to time, and an obvious valley of the apoptotic curve was at the 10th hour after radiation. Compared with the outcome of Western blotting, the results indicated a negative correlation between the apoptotic rate and the radiant intensity or p65 protein expression(P<0.05). CONCLUSIONS: The expression of p65 protein is affected by the irradiation of p65 of high energy X-ray, which is dose-time dependent. The proportion of apoptotic cells decreases as the expression increases.

Hepatocyte growth factor and granulocyte colony-stimulating factor form a combined neovasculogenic therapy for ischemic cardiomyopathy.

BACKGROUND: Myocardial infarction (MI) is a significant cause of heart failure. Current therapies are limited and, therefore, the development of novel revascularization methods is potentially important. We investigated whether hepatocyte growth factor (HGF), expressed by genetically modified mesenchymal stromal cells (MSC), in combination with granulocyte colony-stimulating factor (G-CSF), exhibited a synergistic therapeutic benefit, as measured 8 weeks after MI induction in a rat model. METHODS: Four weeks after MI, rats were randomly divided into a control group (n=11), HGF group (Adenovirus vector carrying human HGF (Ad-HGF)-transfected MSC transplanted into the infarct zone; n=11), G-CSF group (intraperitoneal injection with G-CSF; n=11), and HGF + G-CSF group (Ad-HGF-transfected MSC transplanted into the infarct zone and intraperitoneal injection with G-CSF; n=11). Four weeks later, hearts were analyzed for endothelial cell density and angiogenesis, ventricular geometry, myocardial function and levels of VCAM-1 and MMP-9 protein. RESULTS: The HGF + G-CSF group exhibited improved left ventricular systolic and diastolic function and experienced less adverse ventricular remodeling, as manifested by decreased left ventricular dilatation and increased border zone wall thickness. Angiogenesis was significantly enhanced in HGF + G-CSF rats by inducing the proliferation of endothelial cells. Furthermore, HGF induced expression of VCAM-1, and HGF treatment together with G-CSF synergistically stimulated MMP-9 expression in ischemic hearts. DISCUSSION: The combination of G-CSF and HGF exhibited a significant synergistic effect and enhanced myocardial endothelial density, angiogenesis, geometric preservation and heart function in an ischemic cardiomyopathy model.

[Relationship between nuclear factor kappa B signal transduction way and the sensitivity of tongue carcinoma cell to Pingyangmycin chemotherapy].

OBJECTIVE: To examine the relationship between NF-kappaB signal transduction way and the sensitivity of Tca8113 carcinoma cell to Pingyangmycin chemotherapy. METHODS: 2 mg/L antisense p65 oligodeoxynucle-otides (5'-GAACAGTTCGTCCATGGCCG-3') was transfected into Tca8113 cells through Lepofectin vectin, then the cells were treated with 8 mg/L Pingyangmycin. After 3 h and 6 h, the expression of p65 in nucleus was studied with immunohistochemical methods and Western blot analyses. After 48 h, the inhibitory rate of cell growth was detected with MTT assay. RESULTS: The NF-kappaB/p65 signal transduction way in Tca8113 cells was activated after treated with Pingyangmycin. But the transfection of antisense p65 oligodeoxynucle-otides inhibited the activation of the signal transduction, the p65 expression in nucleus was decreased significantly (P < 0.05) at 6 h, and the inhibitory rate of cell growth was increased significantly (P < 0.05) at 48 h. CONCLUSION: The results suggested the chemotherapeutic sensitivity of Tca8113 carcinoma cell to Pingyangmycin was improved by the inhibition of NF-kappaB/p65 signal transduction.

[High expression of p65 transcription factor and its significance in tongue carcinoma].

OBJECTIVE: To examine the correlation between p65 expression and the metastasis and prognosis in tongue carcinoma. METHODS: The immunohistochemical expression of p65 was examined in 50 cases of tongue carcinoma, and the clinical and histopathologic records were retrospectively analyzed. RESULTS: Overexpression of p65 in cytoplasma and nucleus was found in 64% cases of tongue carcinoma. There was significant difference of p65 expression among different TNM stages, and the higher expression rate of p65 was found in higher stages. Overexpression of p65 was also related to lymph node metastasis. Significantly greater increase in expression levels of p65 was found in the metastatic cases (n = 22) compared to the nonmetastatic cases (n = 10). However, there was no significant difference of p65 expression among different histopathologic grading. The 5-year survival rate of p65 positive and negative group was 59.3% and 88.9% respectively, and there was significant difference in Kaplan-Meier survival curve (chi2 = 5.6, P < 0.05). CONCLUSION: The results suggest that the expression of p65 was important to predict the metastasis and prognosis in tongue carcinoma.