OBJECTIVETo investigate the correlation between methylation status and gene expression of APC (adenomatous polyposis coli) gene in HeLa, CaSki and SiHa cell lines of cervical carcinoma, and explore the effect of hydralazine on the transcription regulation of the 5'CpG island demethylation of APC gene and the proliferation and apoptosis of the cell lines.

METHODSMethylation status and the expression of APC gene were analyzed using methylated specific PCR, RT-PCR and FQ-PCR methods. The expression of beta-catenin protein which correlates closely with APC was detected by SP method after treatment with Hydralazine. MTT and FCM assays were used to observe the changes of proliferation activity and apoptosis of the cells after Hydralazine treatment.

RESULTS(1) APC gene was methylated or hemimethylated respectively in HeLa and CaSki cell lines, at the same time, APC gene was not methylated in SiHa cell. (2) After having been treated by 40 micromol/L Hydralazine for 72 hours, growth inhibitory ratios of HeLa, CaSki and SiHa cell lines were (52.12 +/- 3.78)%, (44.31 +/- 2.59)% and (47.73 +/- 4.73)% respectively, on the contrary, normal cell HECV's growth inhibitory ratio was only (27.18 +/- 0.79)%. APC gene in HeLa and CaSki cell lines which were treated by 40 micromol/L Hydralazine for 72 hours was demethylated and expressed positively, the expression of APC mRNA in HeLa, CaSki and SiHa cell lines increased to 10.35, 11.40 and 0.73 times respectively. (3) Hydralazine, when used at the concentration of 40 micromol/L for 72 hours, induced S phase and G2/M phase arrest and apoptosis in HeLa and CaSki cells. beta-catenin protein can be expressed in cell membrane after treatment with Hydralazine.

CONCLUSIONAPC gene methylation plays an important role in the carcinogenesis of cervical cells and can re-express after the treatment with Hydralazine which also could inhibit the growth of the cervical cancer cells.

Adenomatous Polyposis Coli Protein ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; CpG Islands ; genetics ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Genes, APC ; HeLa Cells ; Humans ; Hydralazine ; pharmacology ; RNA, Messenger ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology ; beta Catenin ; metabolism

OBJECTIVETo investigate the prevalence of high-risk human papillomavirus (HPV) and incidence of cervical intraepithelial neoplasia (CIN) in female populations in Shenzhen, Guangdong Province, China.

METHODSTotally 1137 women aged 15-59 from Shahe Community, Nanshan District, Shenzhen were investigated for cervical cancer during an population-based epidemiological screening from November 2004 to December 2004. Visual inspection with acetic acid (VIA), colposcopy, liquid-based cytology test (LCT), and hybrid capture 2 (HC-) were performed to detect the high-risk HPV types in cervical secretions. Biopsy under colposcope was performed in women who were HPV-positive with LCT >or= atypical squamous cells of undetermined sign (ASCUS) or HPV-negative with LCT >or= low grade squamous intraepithelial lesion (LSIL), with the pathological results as the golden standards.

RESULTSThe detection rate of high-risk HPV-DNA was 14.0%. HPV detection rates in 15-24, 25-29, 30-34, 35-39, 40-44, 45-49, and 50-59 age groups were 15.5%, 17.7%, 12.6%, 8.8%, 10.2%, 15.3%, and 21.0%, respectively (P < 0.05). HPV detection rates in 25-29 years group and 50-59 years group were significantly higher than those in other groups (P < 0.05) and 35-39 group had the lowest detection rate. The curve of HPV infection rates in all groups was 'V' type. The overall incidence of CIN was 4.4%. The incidences of CIN , CIN , and CIN were 3.2%, 1.0%, and 0.3%, respectively, in which the incidence of CIN was significantly higher than those of CIN and . HPV detection rates increased with cervical lesion grades, which in >or=CIN groups and normal group were 100.0% and 8.3%, respectively. No cervical cancer was identified in this research. The sensitivities of VIA, colposcopy, LCT, and HC-II for high-risk HPV screening were 35.7%, 50.0%, 92.9%,and 100%, respectively, in detecting high-grade squamous intraepithelial lesion (HSIL), the specificities of these four methods were 96.0%, 87.2%, 88.4%, and 86.9%, respectively. Satisfactory negative predictive values were obtained for all methods.

CONCLUSIONSHPV infection is the main risk factor for CIN. Cervical cancer among female populations in Shenzhen is still in early stages. Prevention of HPV infection and treatment of CIN are key for the prevention of cervical cancer.

Adolescent ; Adult ; Cervical Intraepithelial Neoplasia ; epidemiology ; Chi-Square Distribution ; China ; epidemiology ; DNA, Viral ; isolation & purification ; Female ; Humans ; Incidence ; Mass Screening ; methods ; Middle Aged ; Papillomaviridae ; genetics ; Papillomavirus Infections ; epidemiology ; Uterine Cervical Neoplasms ; diagnosis ; Young Adult

OBJECTIVETo explore the potential anti-inflammatory and analgesic activities of carboxyamidotriazole (CAI).

METHODSA variety of animal models, including the croton oil-induced ear edema, the cotton-induced granuloma, the rat adjuvant-induced arthritis, were used to evaluate anti-inflammatory effect of CAI. Vascular endothelial growth factor (VEGF)--or histamine-stimulated local vascular permeability in mouse modulated by CAI was also determined. In addition, we assessed the effect of CAI on the levels of proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-beta) at the site of inflammation and in sera. Moreover, antinociceptive effect of CAI on inflammatory pain was assessed using acetic acid-induced writhing model and the formalin test.

RESULTSCAI significantly inhibited acute and chronic phases of inflammation, reduced VEGF or histamine-induced vascular permeability, and showed marked inhibition of proinflammatory cytokines such as TNF-alpha and IL-1 beta. CAI also showed potential therapeutic effect on peripheral inflammatory pain.

CONCLUSIONCAI is a promising anti-inflammatory and analgesic agent.

Analgesics ; pharmacology ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Drug Evaluation, Preclinical ; Female ; Male ; Mice ; Mice, Inbred ICR ; Rats ; Rats, Wistar ; Triazoles ; pharmacology