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Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.
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Objective To detect 5′CpG island methylation of APC(adenomatous polyposis coli) gene promoter region in cervical carcinoma cell lines (HeLa,CaSki,SiHa), and to investigate the possibility of Hydralazine in restoring the expression of APC gene through demethylation. Methods The CpG island methylation status of APC gene promoter region of the cervical carcinoma cell lines were analyzed using methylated specific polymerase chain reaction (MSP). The mRNA expression profile of APC gene was analyzed by using RT-PCR after Hydralazine treatment. Results The CpG island of APC gene was methylated in HeLa and hemimethylated in CaSki but not in SiHa. The mRNA expression of APC gene can be detected after hydralazine treatment. Conclusion The APC gene non-expression in cervical carcinoma is associated with CpG island methylation in APC gene promoter region. Hydralazine, served as a demethylating agent, enables to restore the expression of APC gene.
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Soft environment of opening laboratory in key subject will deeply affect the construction and development of disciplines.Developing and innovative academic team;scientific and rigorous running mechanism,healthy and concordant academic atmosphere,active and positive behavior mode,all of the above are the main factors of soft environment of laboratory.From now on,we need try our best to improve human rights,design more efficient aims,constitute impellingly guarantee system and fine mechanism of consistent of regulations and environment,apply modern managing instrument scientificly and improve managing efficiency.
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Objective To investigate the expression of survivin, matrix metalloproteinase (MMP-2) and tissue inhibitor 2 of matrix metalloproteinase (TIMP-2) in cervical carcinoma and their relationship with invasion and node metastasis of the cervical cancerous tissues. Methods The expressions of survivin, MMP-2 and TIMP-2 were examined by immunohistochemical S-P method and colour pathological image computer analysis system in 10 cases of normal cervical epithelia, 10 cases of cervical carcinoma in situ, 40 cases of invasive squamous cell cervical carcinoma and 11 cases of invasive cervical adenocarcinoma. The relationship between those indexes and the factors related to clinical pathology of cervical carcinoma were analyzed statistically. Results It was found that the positive level of survivin and MMP-2 expression increased in the order of normal cervical epithelium, cervical carcinoma in situ and invasive carcinoma of cervix (P0.05). The positive expressions of survivin and MMP-2 in patients under 35 years old or with pelvic lymph node metastasis, intravascular involvement and stroma involvement were significantly higher than that in the cases without them, while TIMP-2 expression was opposite to that of MMP-2 (P