RESUMEN
OBJECTIVE@#To explore the mechanism of paclitaxel on the protein expression of human cervical carcinoma cell line HCE1.@*METHODS@#The total proteins extracted from paclitaxel-treated HCE1 cells were analyzed by 2-dimensional gel electrophoresis (2-DE), and compared with those from untreated HCE1 cells. The differential proteins were identified by mass spectrometry. Western blot was used to determine the differential expression levels of the 2 proteins.@*RESULTS@#At 24 hour after paclitaxel (0.05 mumol/L) treatment, 2-DE images of paclitaxel-treated and paclitaxel-untreated cells were analyzed. Forty-two differential proteins were found. Twenty-one differential proteins among 42 proteins were analyzed by mass spectrometry, among which 15 proteins were identified, including peptidyl-prolylisomerases A (PPIase A),alpha-enolase,keratin 8,heat shock protein 90, eukaryotic translation initiation factor 1A, and so on.@*CONCLUSION@#Fifteen proteins in human cervical carcinoma cells paclitaxel-treated and paclitaxel-untreated are found by proteomic techniques. These proteins may be involved in the proliferation inhibition of human cervical carcinoma cells by paclitaxel.
Asunto(s)
Femenino , Humanos , Antineoplásicos Fitogénicos , Farmacología , Biomarcadores de Tumor , Proteínas de Unión al ADN , Factor 1 Eucariótico de Iniciación , Perfilación de la Expresión Génica , Genoma , Queratina-8 , Paclitaxel , Farmacología , Fosfopiruvato Hidratasa , Proteoma , Proteómica , Métodos , Células Tumorales Cultivadas , Proteínas Supresoras de Tumor , Neoplasias del Cuello Uterino , Metabolismo , PatologíaRESUMEN
OBJECTIVE@#To investigate the association between the expression of caveolin-1(CAV-1) and the invasion of choriocarcinoma, and to explore the effect of CAV-1 small interfering RNA(siRNA) on the invasion of choriocarcinoma cell line JEG-3.@*METHODS@#(1) Matrigel invasion assay and 3-(4,4)-dimethylthiahiazo (-z-yl)-3,5-di-phenytetrazoliumormide (MTT) assay were used to examine the difference in invasion and proliferation ability between JEG-3 cells and JAR cells;(2) Expression of caveolin-1 gene in the human chorionic villi tissues and chorionicnoma cell lines (JEG-3 cells and JAR cells) were detected by semi-quantitative RT-PCR. (3) The effect of CAV-1 siRNA transfection on the expression of CAV-1 mRNA, and the invasion and proliferation ability of JEG-3 cells were measured by RT-PCR, Matrigel invasion assay and MTT assay.@*RESULTS@#(1) The invasion ability of JEG-3 cell line was stronger than that of JAR cell line (P0.05);(2) The expression of caveolin-1 gene in chorionicnoma cell lines was significantly stronger than that in the human normal chorion(P<0.05), and the expression of caveolin-1 gene in JEG-3 cells was stronger than that in the JAR cells (P<0.05). The data suggested that there was significantly positive correlation between caveolin-1 and the invasiveness of chorionicnoma cells (r=0.086,P<0.05);(3) CAV-1 siRNA could knock-out the expression of CAV-1 mRNA, and inhibit the invasion and proliferation ability of chorionicnoma cells.@*CONCLUSION@#CAV-1 can promote the invasion ability of chorionicnoma cells. CAV-1 siRNA can inhibit the invasion and proliferation ability of chorionicnoma cells.
Asunto(s)
Femenino , Humanos , Caveolina 1 , Genética , Coriocarcinoma , Metabolismo , Patología , Invasividad Neoplásica , Interferencia de ARN , ARN Mensajero , Genética , ARN Interferente Pequeño , Genética , Células Tumorales Cultivadas , Neoplasias Uterinas , Metabolismo , PatologíaRESUMEN
OBJECTIVE@#To explore the influence of carbon dioxide pneumoperitoneum-laparoscopic surgery on tumor cell seeding and metastases in endometrial cancer.@*METHODS@#Twenty patients with endometrial cancer who underwent laparoscopic surgery and 10 patients with endometrial cancer who underwent laparotomic surgery were enrolled. Each patient was in preoperative clinical StageIand the uterus size in each patient was less than 12 weeks of pregnancy. Carbon dioxide pneumoperitoneum was established and maintained with CO2 insufflation at 4 approximately 6 L/min and intraperitoneal pressure of 13 mmHg with an automatic pneumoperitoneum machine. Cytologic examination of peritoneal fluid(at the beginning and end of the operation), CO2 filtrated gas and the lavage fluid of instruments during the laparoscopic surgery were performed. The protein expressions of E-cadherin,beta-catenin,P-selectin,matrix metalloproteinase-2(MMP-2),vascular endothelial growth factor (VEGF),and CD44v6 in tumor tissues before and after the operation were detected by DAKO Envision.@*RESULTS@#There were no case of positive washing cytology in the peritoneal fluid,CO2 filtrated gas, and the lavage fluid of instruments during the laparoscopic surgery. The expressions of E-cadherin and beta-catenin proteins were obviously abnormal in endometrial cancer. The abnormal expressions of E-cadherin and beta-catenin protein between the pre- and post-operations were not significantly different in both the laparoscopic group and the laparotomic group(P>0.05).The changes of abnormal expressions of E-cadherin and beta-catenin protein were no statistical difference between the two groups(P>0.05). The positive protein expressions of P-selectin,MMP-2,VEGF,and CD44v6 were not significantly different between the pre- and post-operations both in the laparoscopic group and the laparotomic group(P>0.05),and there was also no significant difference between the laparoscopic group and the laparotomic group(P>0.05).The follow-up period in the laparoscopic group was 7 approximately 19 (14.25+/-3.65) months and 7 approximately 19 (13.10+/-4.23) months in the laparotomic group. One patient got infection in the urinary system in the laparoscopic group and one patient had lower extremity venous thrombosis in the laparoscopic group.No recurrence was detected in both groups.@*CONCLUSION@#Laparoscopic surgery for endometrial cancer has no effect on protein expressions of E-cadherin,beta-catenin,P-selectin,MMP-2,VEGF,and CD44v6 in tumor tissues. No evidence has been found that CO2 pneumoperitoneum-laparoscopic surgery may favor endometrial cancer cell seeding and metastases.
Asunto(s)
Adulto , Femenino , Humanos , Persona de Mediana Edad , Dióxido de Carbono , Carcinoma Endometrioide , Cirugía General , Neoplasias Endometriales , Cirugía General , Laparoscopía , Metástasis de la Neoplasia , Siembra Neoplásica , Neumoperitoneo ArtificialRESUMEN
<p><b>OBJECTIVE</b>To identify proteins associated with growth inhibitory effects of progesterone in Ishikawa endometrial adenocarcinoma cell line (Ishikawa).</p><p><b>METHODS</b>The total cellular proteins of the control and megestrol acetate (MA)-treated Ishikawa cells were separated by two-dimensional gel electrophoresis. After colloidal Coomassie G-250 staining and scanning, the gel images were analyzed by PDQuest software. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and Mascot software were used to identify the differentially expressed protein.</p><p><b>RESULTS AND CONCLUSIONS</b>Well-resolved and reproducible 2-dimensional patterns of both control and MA-treated Ishikawa cells were obtained. Forty-one proteins displayed differential expression after MA treatment, among which 23 were up-regulated and 18 down-regulated. Peptide mass fingerprints (PMF) by MALDI-TOF-MS analysis and search in NCBInr resulted in the identification of 8 proteins. The up-regulated proteins identified were catechol-O-methyltransferase (COMT), glutathione S-transferase, keratin 8, splicing factor 3a (subunit 3), chaperonin containing TCP1 (subunit 6A isoform a, CCT6A), RAB6A protein and AHA1. The down-regulated protein was peptidyl prolyl isomerase A (isoform 1). These findings can be helpful in further investigation of the molecular mechanism of progesterone.</p>
Asunto(s)
Femenino , Humanos , Adenocarcinoma , Metabolismo , Patología , Línea Celular Tumoral , Regulación hacia Abajo , Electroforesis en Gel Bidimensional , Neoplasias Endometriales , Metabolismo , Patología , Espectrometría de Masas , Acetato de Megestrol , Farmacología , Progesterona , Farmacología , Proteínas , Proteoma , Regulación hacia ArribaRESUMEN
OBJECTIVE@#To determine the effect of progesterone on the secretion of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in ectopic endometrial stromal cells.@*METHODS@#Ectopic endometrial stromal cells were obtained from 17 patients with endometriosis. Endometrial stromal cells were obtained from 12 patients with endometriosis and 14 cases of controls. Ectopic endometrial stromal cells of 15 cases were treated with progesterone. Culture supernatants of these stromal cells were analyzed for MMP-2 and MMP-9 by zymography.@*RESULTS@#Endometriotic stromal cells released significantly higher levels of MMP-2 and MMP-9 than endometrial stromal cells from women with and without endometriosis. Progesterone at 10(-9) mol/L caused endometriotic stromal cells a significant reduction MMP-2 and MMP-9 levels. When progesterone concentration was increased from 10(-9) mol/L to 10(-7) mol/L, the release of MMP-9 was almost completely inhibited, wherease that of MMP-2 was not completely inhibited.@*CONCLUSION@#Progesterone may inhibit the secretion of MMP-2 and MMP-9 in ectopic endometrial stromal cells, especially MMP-9.