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To investigate the expressions of p53 and PCNA by inhibition of hTERT with siRNA in xenografted laryngeal squamous carcinoma in nude mice.pshRNA involved fluorescein gene was synthesized according to the sequence of hTERT cDNA. The xenografted tumor in nude mice was established by using human laryngeal squamous carcinoma cell line (Hep-2). pshRNA packaged by METAFECTENE was transfected into the transplantation tumors. The volumes of the subcutaneous tumor were determined. Fluorescence expression in the tumors was detected by confocal microscopy. The expressions of p53 and PCNA were detected using immunohistochemistry.The subcutaneous tumors appeared in all the nude mice after transplantation five days later. The diameter of subcutaneous tumors become 5-7 mm after transplantation. Many green fluorescent cells were observed by confocal microscopy after transfection of pshRNA and blank plasmid. The growth of laryngeal squamous carcinoma in nude mice was significantly decreased as compared with the control group. The proliferation activity was inhibited by 76.50%. The expressions of p53 was upregulated and the expression of PCNA was downregulated.The transfection of pshRNA could significantly induce inhibitory effects on growth of laryngeal squamous carcinoma in nude mice and the expression of PCNA could be effectively downregulated. The inhibition of telomerase reverse transcriptase (hTERT) mRNA could upregulate the expression of p53 and promote the apoptosis of carcinoma cells as well as inhibit the proliferation of laryngeal squamous carcinoma cells.

To investigate the effect of inhibiting human telomerase reverse transcriptase (hTERT) on expression of C-myc protein by RNA interference (RNAi) in the larynx cancer cell line, Hep-2.The primary structures of hTERT cDNA were found in GeneBank. Then the structure analyses were done according to the strategy of RNAi, which determined the specific base sequences to design shRNA plasmid. One type of plasmid, pshRNA1, involved in fluorescein gene was synthesized based on the specific base sequence. Control pshRNA2-a random sequence-were also constructed. METAFECTENE was used as the transfect ion reagent. Cells were treated daily with pshRNA1-2 or normal culture medium respectively. After administration of pshRNA1-2, hTERT mRNA was detected by RT-PCR, hTERT protein and C-myc protein were examined by Western Blot.The expression of hTERT mRNA and protein were both significantly decreased after treated by pshRNA1 (P0.05). The expression of C-myc protein was significantly increased after treated by pshRNA1 (P0.01).The inhibition of hTERT expression could increase the expression of C-myc protein in Hep-2 cells.

To investigate nuclear factor-kappaB/p65(NF-kappaB/p65) and cyclooxygenase-2 (COX-2) gene expressions and clinical significance in human laryngeal squamous cell carcinoma, at the same time to investigate the relationship between the two gene expressions.Forty-one samples of laryngeal squamous cell carcinoma and ten samples of vocal cord polyps were assayed using immunohistochemical SP method for detection of NF-kappaB/p65 and COX-2 protein expression level.There is significant difference between the laryngeal carcinoma group and vocal cord polyps group about NF-kappaB/p65 and COX-2 protein expression level. Qualitative analysis found theres significant relationship between COX-2 and histological classification. There's significant correlation relationship between NF-kappaB/p65 and COX-2 by using spearman correlation analysis.(1)The expressions of NF-kappaB/p65 and COX-2 both raise in laryngeal carcinoma. (2) NF-kappaB/p65 perhaps promotes the expression of COX-2 in laryngeal carcinoma. (3) NF-kappaB was hoped to become the new treated target of laryngeal carcinoma.

To investigate the photodynamic therapy (PDT) effect of a new kind of cationic porphyrin on human laryngeal cancer Hep-2 cell in vitro.A new cationic porphyrin (5-(4-methoxycarbonylphenyl)-10, 15, 20-tris(4-N-methylpyridiniumyl)porphyrin triiodide ) was synthesized. The Hep-2 cells were treated with three different concentrations of porphyrin (10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L) followed by irradiation of different visible light (0, 0.5, 1.0, 1.5, 2.0 h). MTT and TUNEL were applied to measure the inhibitory effect of porphyrin on the Hep-2 cells growth, inhibitory rate and induced apoptosis rate.Under definite condition, the cantionic porphyrin concentrations had a significant inhibitory effect and induced apoptosis effect. The effect depended greatly on the doses of the concentration and the time of irradiation of light. At the highest concentration of porphyrin (10(-5) mol/L), the growth inhibitory rate were 1.18%, 35.21%, 58.59%, 70.42%, 84.34% respectively and the apoptosis rate were 3.30% +/- 0.29%, 29.41% +/- 2.12%, 32.30% +/- 2.26%, 34.52% +/- 3.11% in different visible light (0.05, 1.0, 1.5, 2.0 h), respectively. Under high irradiation energy, it can causesome Hep-2 cells death directly.Porphyrin has a distinctive killing effect on Hep-2 cells and correlated with the doses and the irradiation time.

To observe the distribution of gama-aminobutyric acid (GABA) ergic neurons in rats cochlear nuclei (CN) in pre-and post-operation of unilateral cochlea ablation.The technique of direct anti-GABA immunocytochemistry (SP) was used in this study.The GABAergic neurons existed extensively in cochlear nuclei(CN). The number was more, the volume was larger of GABAergic cells in CN and those cells were stained heavily before ablation. The number of GABAergic cells in lesion side was far less than that in intact side after unilateral cochlea ablation one to two weeks (P0.01), then the number of GABAergic cells increased after three weeks which was still less (P0.05) and almost reached normal after a month (P0.05).The changes of GABAergic neurons reflect the balance between excitatory and inhibitory of neurons. It suggests that changes in the extent of GABAergic inhibition have been shown to contribute to the reorganization of primary auditory central system after unilateral cochlea ablation in rats. The decrease and increase of the number of GABAergic-neurons are essential for this reorganization.

To observe the effects of systemic steroids on the infiltration of eosinophils and the expression of TGF beta 1 in nasal polyps.The SP immunohistochemical methods were applied to explore the expression of TGF beta 1 in nasal polyps before and after corticosteroids use; the optical density of positive cells in mucosal epithelia, eosinophils, glandular epithelium, vascular endothelial, matrix and mucous membrane was measured by using HPIAS-1,000 image-conduct system.The expression of TGF beta 1 was positive in nasal polyps. The number of eosinophils and the expression of TGF beta 1 was significantly reduced in nasal polyps after treated with corticosteroids compared with pre-treated.The effects of systemical steroid use on the nasal polyps may depend on decreasing the infiltration of eosinophils and the expression of TGF beta 1.

To study the effect of antisense oligonucleotide targeted human telomerase catalytic subunit (hTERT) on Hep-2 cell line.Cells were treated daily with oligonucleotide (ODN) directed against hTERT, or control ODN-a mismatched sequence or only Lipofectamine. At 24 h, 48 h, 72 h cell viability after administration of ODNs was determined using the MTT assay. At 72 h, the morpholog alteration of the cells was observed by HE stain. At 72 h, telomerase activity was determined.After daily transfection ASODN, some Hep-2 cells begin to stop growing up, but not in the mismatched ODN group or Lipofectamine group. Hep-2 cells treated with antisense against hTERT showed a decrease in telomerase activity.hTERT antisense ODN can inhibit telomerase activity and the growth of Hep-2 cells in short term.