OBJECTIVE: Human cervical cancer is caused by the high-risk types of human papillomavirus (HPV) such as HPV16, which possess the E6 and E7 oncogenes, whose expressions are a prerequisite for cancer development. We performed this study to compare the efficacy of antitumor activity by HPV siRNA which silences only E6 or both E6/E7. METHODS: We transfected siRNA 377 (HPV16 E6 siRNA), siRNA 3 (HPV16 E6 siRNA), and siRNA 198 (HPV16 E7 siRNA) into SiHa cell line (siRNA 377 silences only E6, and siRNA 3 and siRNA 198 silence both E6 and E7). We experimented cell counts and morphologic changes 24 and 48 hours after transfection and expressions of HPV16 E6/E7 mRNA by RT-PCR. RESULTS: siRNA 377, siRNA 3, and siRNA 198 suppressed the cell growth. siRNA 3 and siRNA 198 were more potent than siRNA 377 in cell growth suppression. siRNA 377 knocked down the expression of E6 mRNA, and both siRNA 3 and siRNA 198 knocked down the expression of E6/E7 mRNA. CONCLUSION: Our results suggest that simultaneous suppression of E6 and E7 was more potent than E6-specific suppression in cancer cell growth.
Cell Count ; Cell Line ; Humans ; Oncogenes ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Uterine Cervical Neoplasms

No abstract available.

Objectives of this study were to establish a leukemia mouse model in the Balb/c mouse based upon the A20 cell line (murine B-lymphoma/leukemia cell line, H-2d). Here we demonstrate for the first time that A20 cells were infiltrated into tissue and bone marrow, thereby evaluate the feasibility of using A20 leukemic cells as a leukemia model. In the study, changes of behavior, survival rate and histological changes of major organs after intravenous injection of A20 cells (1x105, 1x106 or 1x107) into Balb/c mice were observed. After inoculation of 1x106 cells, animals survived up to 38.3 days, although there were no significant correlation between the number of injected cells and life-span. At 21 and 28 days post-injection, both hematoxylin-eosin and CD45R immunohistochemical stains showed diffuse large B-cell lymphoma in the liver. FACS analysis was performed after injection of fluorescent nanomaterial (MNPs@SiO2 RITC)-labeled A20 cells. The labeled A20 cells were detected in bone marrow from 6 hours post-inoculation, indicative of the cellular infiltration. This is the first study that demonstrated the invasion of A20 cells into the bone marrow of Balb/c model using A20 cells. With the occurrence of systemic lesions following metastasis of the cells into lymph nodes and neighboring tissues via bone marrow infiltration, it is suggested that the A20 cell-inoculated Balb/c miouse could be an animal model of acute lymphocytic leukemia.
Animals ; Bone Marrow ; Cell Line ; Coloring Agents ; Injections, Intravenous ; Leukemia ; Liver ; Lymph Nodes ; Lymphoma, B-Cell ; Mice ; Models, Animal ; Nanostructures ; Neoplasm Metastasis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Survival Rate

PURPOSE: To maintain the homeostasis of stem cells and prevent their ability to initiate tumorigenesis, it is important to identify and modify factors that prevent or accelerate stem cell senescence. We used microarrays to attempt to identify such factors in human amniotic fluid (HAF)-derived stem cells. MATERIALS AND METHODS: To identify gene expression changes over a time course, we compared gene expression profiles of HAF-derived stem cells in different passages (1st, 2nd, 4th, 6th, 8th, and 10th) using a Sentrix Human illumina microarray. RESULTS: Of the 25,804 genes in the microarray chip, 1,970 showed an over 2-fold change relative to the control (the 1st passage)-either upregulated or downregulated. Quantitative real-time PCR validated the microarray data for selected genes: markedly increased genes were CXCL12, cadherin 6 (CDH6), and folate receptor 3 (FOLR3). Downregulated genes included cyclin D2, keratin 8, insulin-like growth factor 2 (IGF2), natriuretic peptide precursor B (NPPB) and cellular retinoic acid binding protein 2 (CRABP2). The expression pattern of the selected genes was consistent with the microarray data except for CXCL12 and IGF2. Interestingly, the expression of NPPB was dramatically downregulated along the time course; it was almost completely shut-down by the 10th passage. In contrast, FOLR3 mRNA expression was dramatically increased. CONCLUSION: Taken together, although a function for NPPB and FOLR3 in stem cell senescence has not been reported, our results strongly suggest that NPPB and/or FOLR3 play a significant role in the regulation of stem cell senescence.
Aging ; Amniotic Fluid ; Carrier Proteins ; Cell Transformation, Neoplastic ; Cyclin D2 ; Female ; Folic Acid ; Gene Expression ; Homeostasis ; Humans ; Keratin-8 ; Nitrobenzoates ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Stem Cells ; Transcriptome ; Tretinoin

The human papillomavirus (HPV)-16/18 AS04-adjuvanted cervical cancer vaccine has been demonstrated to be highly efficacious and immunogenic with a favorable safety profile. This study assessed the immunogenicity and safety of the HPV-16/18 AS04-adjuvanted vaccine in healthy Korean girls aged 10-14 yr. This multi-center, observer-blind trial randomly assigned 321 healthy girls to receive three doses (0, 1, 6-month schedule) of HPV-16/18 AS04-adjuvanted vaccine or hepatitis A vaccine. Immunogenicity against vaccine antigens was assessed one month post-Dose 3. Solicited and unsolicited adverse events (AEs) and serious AEs (SAEs) were recorded. In the according-to-protocol analysis, all initially seronegative subjects vaccinated with the HPV-16/18 AS04-adjuvanted vaccine had seroconverted at Month 7, with a peak geometric mean titer (GMT) that was 600-fold higher than the natural infection titer of 29.8 EU/mL for HPV-16 and a peak GMT that was 400-fold higher than the natural infection titer of 22.6 EU/mL for HPV-18. The vaccine was well tolerated with no increase in reactogenicity with subsequent doses and no reports of vaccine-related SAEs. In conclusion, the HPV-16/18 AS04-adjuvanted vaccine is shown to be highly immunogenic and generally well-tolerated in Korean girls aged 10-14 yr.
Adjuvants, Immunologic/administration & dosage ; Adolescent ; Aluminum Hydroxide/administration & dosage ; Antibodies, Viral/analysis ; Child ; Female ; Hepatitis A/immunology ; Hepatitis A Vaccines/administration & dosage/adverse effects/immunology ; Humans ; Lipid A/administration & dosage/analogs & derivatives ; Papillomavirus Infections/*prevention & control ; Papillomavirus Vaccines/administration & dosage/adverse effects/*immunology ; Republic of Korea ; Seroepidemiologic Studies ; Uterine Cervical Neoplasms/*prevention & control

BACKGROUND: For the detection of HLA antibodies, solid-phase tests using purified HLA antigens are increasingly used. In this study, we analyzed the panel reactive antibody (PRA) test results using ELISA and Luminex methods, and the results were compared with those of crossmatch test. METHODS: A total of 111 sera including 90 sera from kidney transplanted patients were tested. ELISA-PRA was performed using Lambda Antigen Tray Class I and II Mixed kits (One Lambda Inc., USA) and additional test was performed to identify HLA specificities. Luminex-PRA tests were performed using LABScreen Mixed kits (One Lambda Inc., USA) and LIFECODES LifeScreen Deluxe kits (Tepnel Co., USA). RESULTS: The positive rates of PRA were higher in Tepnel (P=0.006) and One Lambda Luminex (P<0.001) methods than ELISA, without significant difference between two Luminex methods (P=0.087). The overall concordance rate among the three PRA tests was 62.2% (69/111). The positive and negative predictive values of PRA tests for the flow cytometric crossmatch were 33.3-45.7% and 85.7-89.5%, respectively. Of the two Luminex methods, One Lambda showed higher positive rate than Tepnel for the detection of class I antibodies. The sensitivity of pretransplant PRA for the detection of posttransplant acute rejection episodes was higher in Luminex (P=0.007 for Tepnel, P=0.003 for One lambda) than ELISA method. CONCLUSIONS: Different methods used to detect HLA antibodies showed discrepant results. As the Luminex method was more sensitive than ELISA for the detection of HLA antibodies, it can be used as a routine test in the transplantation laboratory.
Enzyme-Linked Immunosorbent Assay/*methods ; Flow Cytometry ; Histocompatibility Antigens Class I/*immunology ; Histocompatibility Antigens Class II/*immunology ; Humans ; Isoantibodies/*blood ; Kidney Transplantation/immunology ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

OBJECTIVE: Arsenic trioxide (As2O3) is known to have potent anti-vascular activity and significantly suppress solid tumor growth. The present study was conducted to investigate the vascular shutdown effects of a novel arsenic compound, tetraasrsenic oxide (As4O6), in comparison with As2O3 using cervical cancer animal model. METHODS: Mice tumor challenge model was used C57BL/6 mice transplanted with TC-1 cells. After the growth of tumors was reached up 200~250 mm3, mice were divided into 3 groups randomly for control and treatment of either As2O3 or As4O6. As2O3 and As4O6 was treated by i.p. injection. The tumor size was caliperated in twice for weeks and anti-vascular effect were assessed by Evans blue extraction assay and Hoechst 33342 staining. In tumor tissue, histopathological feaure was obserevd by hematoxylin and eosin (H&E) staining. RESULTS: In mice treated with either As2O3 and As4O6 (i.p.), both of As2O3 and As4O6 was significantly suppressed the tumor growth compared with control group. Moreover, effect of As4O6 is more pronounced. These tumor growth inhibition is led to the massive necrosis and vacular shutdown in tumor tissue. CONCLUSION: This study suggests that As4O6 may have potential anticancer activity via vascular shutdown in C57BL/6 mice transplanted with TC-1 cells.
Animals ; Arsenic ; Arsenicals ; Benzimidazoles ; Eosine Yellowish-(YS) ; Evans Blue ; Hematoxylin ; Mice ; Models, Animal ; Necrosis ; Oxides ; Transplants ; Uterine Cervical Neoplasms

The aim of present study was to investigate the effects of kaempferol on cellular proliferation and cell cycle arrest and explore the mechanism for these effects in human breast carcinoma MDA-MB-453 cells. Cells were treated with kaempferol at various concentrations (ranging from 1 to 200 microM) for 24 and 48 hrs. Kaempferol significantly inhibited cancer cell growth in cells exposed to 50 and 10 microM of kaempferol and incubated for 24 and 48 hrs, respectively. Exposure to kaempferol resulted in cell cycle arrest at the G2/M phase. Of the G2/M-phase related proteins, kaempferol down-regulated CDK1 and cyclin A and B in cells exposed to kaempferol. In addition, small DNA fragments at the sub-G0 phase were increased by up to 23.12 and 31.90% at 10 and 50 microM incubated for 24 and 48 hrs, respectively. The kaempferol-induced apoptosis was associated with the up-regulation of p53. In addition, the phosphorylation of p53 at the Ser-15 residue was observed with kaempferol. Kaempferol inhibits cell proliferation by disrupting the cell cycle, which is strongly associated with the induction of arrest at G2/M phase and may induce apoptosis via p53 phosphorylation in human breast carcinoma MDA-MB-453 cells.
Apoptosis ; Breast ; Breast Neoplasms ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Proliferation ; Cyclin A ; DNA ; Humans ; Kaempferols ; Phosphorylation ; Proteins ; Up-Regulation

OBJECTIVE: Apigenin is a widely distributed plant flavonoid and was proposed as a potent antitumor agent. In this study, we investigated the anticancer effects of apigenin on human cervical cancer cell lines. For this, the effects of apigenin on growth inhibition and apoptosis were examined and also mRNA expression of E6 and E7, the main causes of development of cervical cancer, was also evaluated. METHODS: To observe the anti-proliferative effects, cervical cancer cell lines, 5x103 cells/well (96 well plate) of Caski, HeLa and C33A were plated and 24 h later treated with apigenin for three days and then MTT assay was performed. For apoptosis analysis, Annexin V-FITC staining was performed. To examine the effect on anchorage-independent growth by apigenin, soft agar assay was performed. The mRNA expression of HPV E6/E7 was examined by quantitative RT-PCR. RESULTS: Apigenin inhibited the growth of all three kind of cervical cancer cell lines (CaSki, HeLa, and C33A) and induced apoptosis in these cell lines. Also, anchorage-independent growth of Caski cells in soft agar was inhibited approximately 3 folds by apigenin treatment. Unexpectedly, mRNA expression level of both E6 and E7 in HeLa cells was not significantly affected by apigenin. CONCLUSION: These studies showed that apigenin inhibits cervical cancer cell growth through the induction of apoptosis. However, mRNA expression of HPV E6/E7 genes was not affected by the treatment of apigenin, indicating that the anti-cancer effect of apigenin in cervical cancer might be mediated via other pathway. Taken together, these results suggest that apigenin may provide a new therapeutic approach for cervical cancer.
Agar ; Apigenin ; Apoptosis ; Cell Line ; HeLa Cells ; Humans ; Plants ; RNA, Messenger ; Uterine Cervical Neoplasms

OBJECTIVE: The discovery of new biomarkers for ovarian cancer is clearly necessary for the detection and monitoring of the disease. Experion(TM) automated electrophoresis system can be employed in the identification of differentially expressed proteins in cancer cells. The objective of this study was to discover potential diagnostic serological biomarkers for ovarian cancer. METHODS: We performed protein expression difference analyses for 14 healthy women and 28 ovarian cancer patients with stage I, III and IV using Experion(TM) system. And then we checked the protein expression as silver staining after loading at 8~16% gradient gel for comparison with Experion(TM) gel image. The candidate biomarkers were purified and determined using MALDI-TOF mass spectrometer. RESULTS: The distinctive polypeptide peaks were detected at 115.40, 15.96, 14.8, 11.66, and 10.69 kDa and these five peaks were identified as ceruloplasmin, hemoglobin beta chain, hemoglobin sigma chain, serum amyloid A4, and amyloid related serum protein SAA, respectively. These proteins were significantly different between the sera of normal healthy women and ovarian cancer patients. CONCLUSIONS: Five proteins were found to be significantly different between the sera of normal healthy women and ovarian cancer patients. In addition, Experion(TM) assay system can provide high performance for analysis of ovarian cancer-related proteins by increasing the throughput while maintaining a high level of accuracy.
Amyloid ; Biomarkers* ; Ceruloplasmin ; Electrophoresis ; Female ; Humans ; Ovarian Neoplasms* ; Silver Staining