ABSTRACT
AIM:To investigate the effect of high-mobility group box-1 (HMGB1) expression knockdown on the invasion ability of breast cancer cells induced by tumor necrosis factor-α (TNF-α).METHODS:HMGB1 siRNA was used to transfect into the breast cancer MDA-MB-231 cells.The expression of HMGB1 at mRNA and protein levels was determined by RT-qPCR and Western blot.After the MDA-MB-231 cells with HMGB1 expression knockdown were treated with TNF-α, the apoptosis rate was analyzed by flow cytometry, the cell invasion ability was measured by Transwell assay, and the cell migration ability was detected by cell scratch test.The protein expression of E-cadherin, MMP-2, N-cadherin, MMP-9 and Bax was determined by Western blot.RESULTS:The expression of HMGB1 at mRNA and protein levels in the MDA-MB-231 cells transfected with HMGB1 siRNA was significantly lower than that in the non-transfected cells (P<0.05).The apoptosis rate in the cells was increased after TNF-αtreatment, and the cell invasion and migration abilities were also increased.The protein level of E-cadherin in the cells was decreased, the protein level of N-cadherin was increased, and the protein levels of MMP-2, MMP-9 and Bax were also increased (P<0.05).After the MDA-MB-231 cells with HMGB1 expression knockdown were induced by TNF-α, the apoptotic rate was increased, the invasion and migration abilities were decreased, the protein levels of E-cadherin and Bax were increased, and the protein levels of N-cadherin, MMP-2 and MMP-9 were decreased, as compared with the cells only induced by TNF-αwithout knockdown of HMGB1 expression (P<0.05).CONCLUSION:Knockdown of HMGB1 expression enhances the apoptosis of breast cancer cells induced by TNF-α, and inhibited the cell invasion, migration and epithelial-mesenchymal transition induced by TNF-α.The mechanism may be related with the changes of protein expression of MMP-2, MMP-9 and Bax.
ABSTRACT
Objective: To investigate the expression of NR4A2(Nurrl) and its splicing variants in gastric cancer, and to evaluate their potential association with the pathogenesis and metastasis of gastric carcinoma. Methods: Alternative Splicing Database (ASD) was used to predict the possible splicing variants of NR4A2; they were identified by gene sequencing technique and their expression was detected by semi-quantitative PCR. The mRNA expression of NR4A2 and its splicing variants in tumor and the corresponding adjacent-tumor tissues (n = 41) were determined by real-time PCR. Meanwhile, immunohistochemisty was employed to examine the protein levels of NR4A2 in the tumor tissues (n = 28). Results: The expression of NR4A2 mRNA was observed in the primary tumor tissues, adjacent tumor tissues, and metastatic tissues. Two splicing variants of NR4A2 were discovered: NR4A2-I and NR4A2-II. The mRNA expression of NR4A2, NR4A2-I, and NR4A2-II in the adjacent-tumor tissue was significantly higher than that in the primary tumor tissue (P = 0.017, P = 0.007, and P = 0.004), and that in the hepatic metastatic tissue was significantly lower than that in the primary tumor tissue (P = 0.001, P = 0.018, P = 0.016). Meanwhile, immunohistochemistry showed that the positive rate of NR4A2 protein was higher in adjacent-tumor tissue than that in both tumor tissues and metastatic tissues, but with no significant differences. Conclusion: At least there are 2 splicing variants of NR4A2 in the gastric tumor, adjacent-tumor, and metastatic tissues; NR4A2 and the novel splicing variants may contribute to the pathogenesis and metastasis of gastric malignancy.
ABSTRACT
Aim To study the actions of Luofendeine and its component. Methods LD50 tests of Luofendeine and its component, heat_board text and body_twisting test of mice were pertormed to learn its analgesic and antiinflammatory actions. Results The analgesic action and antiinflammatory action were increased when codamine was mixed with buluofen by the rate of 13∶200,without increasing its toxicity. Conclution The complex prescription of Luofendeine is rational and can increase the analgesic action obviously.