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Practical Oncology Journal ; (6): 305-309, 2019.
Article in Chinese | WPRIM | ID: wpr-752859


Objective The aim of this study was to investigate the mechanism of sanguinarine(SANG)on the inhibitory pro-liferation in ovarian cancer SKOV3 cells. Methods CCK-8 assay was used to detect proliferation of SKOV3 cells. Flow cytometry was used to detect the effect of SANG on apoptosis in SKOV3 cells. The spectrophotometer was used to detect the production of reac-tive oxygen species(ROS)by SANG. The mouse ovarian cancer xenograft model was used to detect the inhibitory effect of SANG on tumor growth. Results SANG promoted apoptosis in SKOV3 cells in a dose-and time-dependent manner. The SANG-induced ap-optosis was associated with the production of ROS,Activated the c-Jun-N-terminal kinase( JNK) and nuclear factor-κB( NF-κB)signaling pathways. In mouse model of ovarian cancer xenografts,after intravenous injection of mice with SANG,SANG was signifi-cantly inhibited the growth of ovarian cancer xenografts when compared to the control group. SANG also significantly induced apoptosis in ovarian cancer xenografts. Conclusion SANG can significantly inhibit the proliferation of ovarian cancer SKOV3 cells,induce ap-optosis,increase the production of ROS,and inhibit the growth of ovarian cancer.

Article in English | WPRIM | ID: wpr-36485


Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (β-actin), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.

Animals , B-Lymphocytes/parasitology , Cattle , Cell Line , Cells/parasitology , Cells, Cultured , Gene Expression Profiling , Host-Parasite Interactions/genetics , Real-Time Polymerase Chain Reaction/veterinary , Reproducibility of Results , Signal Transduction/genetics , Theileria annulata/physiology , Theileriasis/physiopathology
Article in English | WPRIM | ID: wpr-149765


Triptolide, a compound extracted from the traditional Chinese medicine preparation of Tripterygium wilfordii Hook F., has been reported to have anti-inflammatory and anti-cancer activities. However, its effect on ovarian cancer invasion is unknown. We observed that MMP7 and MMP19 expression increased in ovarian cancer tissue. Triptolide treatment inhibited the migration and invasion of ovarian cancer cells SKOV3 and A2780 at the concentration of 15 nM. We also observed that triptolide suppressed MMP7 and MMP19 promoter activity in a dose-dependent manner, down-regulating the expressions of these promoters on mRNA and protein level. Moreover, triptolide enhanced E-cadherin expression in ovarian cancer cells. In vivo, triptolide inhibited tumor formation and metastasis in nude mice, and suppressed MMP7 and MMP19 expression; it also enhanced E-cadherin expression in tumor in a dose-dependent manner. Over expression of MMP7 and MMP19, or suppression of E-cadherin expression partially abolished the inhibitory effect of triptolide on invasion of ovarian cancer cells. To summarize, triptolide significantly inhibited the migration and invasion of ovarian cancer cells by suppression of MMP7 and MMP19 and up-regulation of E-cadherin expression. This study shows that triptolide is a good candidate for the treatment of ovarian cancer and reduction of metastasis.

Animals , Antineoplastic Agents, Alkylating/pharmacology , Cadherins/genetics , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Cystadenocarcinoma, Serous/drug therapy , Diterpenes/pharmacology , Epoxy Compounds/pharmacology , Female , Gene Expression Regulation, Enzymologic/drug effects , Humans , Matrix Metalloproteinase 7/genetics , Matrix Metalloproteinases, Secreted/genetics , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness , Ovarian Neoplasms/drug therapy , Paclitaxel/pharmacology , Phenanthrenes/pharmacology , Promoter Regions, Genetic , Up-Regulation/drug effects , Xenograft Model Antitumor Assays
Article in Chinese | WPRIM | ID: wpr-383152


Objective To establish a method of nucleic acid extraction and enrichment based on magnetic nanoparticle as medium for elevating the analytical sensitivity of domestic HBV real-time PCR kit and detection of the trace amount HBV DNA. Methods After receiving antiviral treatment, the serum samples of 50 hepatitis B patients with HBV DNA concentration ≤1×104 IU/ml were collected. The WHO HBV DNA calibrator was used as the standard material. Nanometer magnetic beads were used to adsorb and enrich the HBV nucleic acid and increase the concentration of the extracted HBV nucleic acid template. Compared with Roche HBV DNA detection reagent and four domestic reagent with conventional nucleic acid extraction and detection method, the improvement effect of this method on domestic nucleic acid detection reagent was evaluated. Results After application of nanometer magnetic extraction method to domestic regent, the analytical sensitivities of the domestic reagent reached 10 and 50 IU/ml, respectively,which was about the same detection level to 12 IU/ml of the imported Roche reagent. The positive rates of the detection of serum trace amount HBV DNA of hepatitis B patients with four kinds of domestic extraction reagent were 64% ( 32 ), 56% ( 28 ), 62% ( 31 ) and 58% (29), respectively. There were significant statistical differences between Roche reagent and four domestic extraction reagent kits(x2 = 7. 895, 12. 698,9. 013 and 11. 416 ,P <0. 05 ). With nanometer magnetic extraction method combined with domestic reagent kits, the detection rates were 88% (44), 88% (44), 88% (44) and 86% (43) ,respectively. There was no significant difference compared with the imported Roche reagent (x2 = 0. 000, 0. 000, 0. 000 and 0. 088,P >0. 05). Moreover, when the HBV nucleic acid concentration was 101-103 IU/ml, the logarithm value of viral nucleic acid concentration was in reverse correlation to Ct value, but the correlation decreased in the concentration range of 103-106 IU/ml. Conclusions The nucleic acid extraction method based on magnetic nanoparticle as medium can significantly improve the analytical sensitivity of domestic HBV DNA detection reagent, which can be used to monitor the trace amounts HBV DNA in the sera of the hepatitis B patients.