ARHGAP1 overexpression inhibits proliferation, migration and invasion of C-33A and SiHa cell lines.

ARHGAP1, also known as RhoGAP, RhoGAP1, CDC42GAP and p50rhoGAP, is officially named Ras homology (Rho) GTPase-activating protein 1, which is one of the key members of RhoGAPs. Growing evidences demonstrate that several RhoGAPs are suppressed or downregulated in cancers. Thus, the aim of this study was to explore the effects of ARHGAP1 on cervical carcinoma cells. The human cervical carcinoma cells C-33A and SiHa were transduced with lentivirus targeting ARHGAP1 (lenti-ARHGAP1). Cellular proliferation, migration and invasion assays, as well as quantitative real-time polymerase chain reaction and Western blot assays, were performed in the control, negative control (infected with lentivirus) and ARHGAP1+-infected groups. Results showed that overexpression of ARHGAP1 markedly inhibited the proliferation of both C-33A and SiHa cells at 24 h, 48 h and 72 h in a time-dependent manner (n=3, P<0.01). Migration and invasion of C-33A and SiHa cells were suppressed after the transduction with lenti-ARHGAP1 compared with the controls (n=3, P<0.01). In addition, several tumor cellular process-related proteins, such as matrix metallopeptidase 2, zinc finger E-box binding homeobox 1, Cyclin B1, twist family bHLH transcription factor 1 and proliferating cell nuclear antigen, were all downregulated in ARHGAP1-overexpressed C-33A and SiHa cells and proved to be targets of ARHGAP1. This study indicated that ARHGAP1 may have a positive function on antitumor activity in the treatment of cervical cancer.

[Risk of cervical cancer and precancerous diseases in the oral HPV carriers].

OBJECTIVE: To evaluate the risk of the occurrence of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer in the oral HPV carriers through a population-based investigation in Shanghai. METHODS: A total of 1200 cases of outpatients who attended the annual cervical examination and 50 preoperational cases of inpatients with CINIII or invasive cervical cancer were enrolled from three clinical centers in Shanghai. The oral HPV infection was determined by real-time PCR. In 1200-case cross-sectional study, the incidence rate of CIN was compared between the oral HPV positive and negative cohort. In 1250-case case-control study, the positive rate of oral HPV DNA test was compared among normal control group, CINI-III, and invasive cancer case groups, and all odds ratio (OR) values were calculated, respectively. The HPV transmission-related demographic and behavioral characters of the oral HPV carriers were also analyzed. RESULTS: The oral HPV carriers accounted for 5.9% (71/1200) of the investigated outpatients. The oral HPV DNA positive rates were gradually increased with the cervical disease grades, which were 5.8% (68/1182, normal), 2/13 (CIN I), 1/5 (CINII), 31.4% (11/35, CINIII) and 5/15 (invasive cancer). In cross-sectional cohort studies, the relative risks (RR) of CINI,II were 2.9 and 4.0 for oral HPV carriers, respectively. In case-control study, the OR values for CINI-III and invasive cervical cancer were 3.1(95%CI: 1.6-10.1), 4.2(95%CI: 1.7-28.4), 7.1(95%CI: 4.8-19.8) and 10.1(95%CI: 3.2-32.1), respectively. The oral sex and multi-sexual partner were two major risk factors for the oral and cervical HPV co-infection, HPV-related cervical cancer and precancerous diseases according to behavioral analysis. CONCLUSIONS: There are complicated transmission pathways between oral and cervical HPV. Oral HPV carriers should be intensively followed up and their oral HPV infection and HPV-related cervical diseases should be treated together.

Preparation, characterization and in vitro release study of a glutathione-dependent polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan.

In this work, an amphiphilic polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan (PTA-g-CMCS) was designed and synthesized. In aqueous solution, this grafted polymer can self-assemble into spherical micelles with a size ranging from 104 to 285 nm and zeta potential ranging from -12.3 to -20.1 mV. For the release study, less than 24% of 6-Mercaptopurine (6-MP) was released from PTA-g-CMCS1 in the media containing 2 and 100 µM glutathione (GSH), whereas 37%, 54% and 75% of 6-MP was released from the media with GSH of 1, 2 and 10mM, respectively. Besides, pH and drug content of the polymeric prodrug only presented slight influence on the 6-MP release. MTT assay demonstrated that this system had higher inhibition ratio on HL-60 cells (human promyelocytic leukemia cells) in the presence of GSH and lower cytotoxicity on mouse fibroblast cell line (L929). Therefore, this nano-sized system is glutathione-dependent, and it can be employed as a potential carrier for the controlled release of 6-MP.