Effect of Cynanoside H on proliferation and metastasis of triple negative breast cancer MDA-MB-231 cells and its mechanism
- VernacularTitle:Cynanoside H对三阴性乳腺癌MDA-MB-231细胞增殖和转移的影响及其机制
- Author:
ZHAO Peng
- Publication Type:Journal Article
- Keywords:
Cynanoside H;
Triple negative breast cancer(TNBC);
MDA-MB-231 cells;
Cell proliferation;
Cell metastasis
- From:
Chinese Journal of Biologicals
2024;37(10):1190-1199
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Cynanoside H on the proliferation and metastasis of triple negative breast cancer(TNBC) MDA-MB-231 cells and its mechanism, so as to provide an experimental basis for the development of TNBC therapeutics.Methods MDA-MB-231 cells were randomly divided into one control group(without Cynanoside H) and three test groups including 2. 5, 5 and 10 μmol/L Cynanoside H dose groups. The effect of Cynanoside H on the proliferation of MDA-MB-231 cells was detected by MTT assay and clone formation assay, while the effect on the cell cycle was detected by flow cytometry, and the effect on the metastasis of MDA-MB-231 cells was measured by wound healing assay, Transwell migration and invasion assay. In addition, the effects of Cynanoside H on the expression of cell cycle(c-Myc, CDK1, CDK2and cyclin E1), metastasis(E-cadherin, Vimentin and β-catenin) and PDGFRB/JAK2/STAT3 pathway related proteins(PDGFRB, p-JAK2, JAK2, p-STAT3 and STAT3) in MDA-MB-231 cells were determined by Western blot.Results The levels of cell viability(t = 4. 598, 19. 77 and 53. 43, respectively, each P < 0. 05), growth(36 h: t = 8. 256, 11. 57 and12. 16, respectively, each P < 0. 05; 48 h: t =10. 49,22. 49 and 19. 63, respectively, each P < 0. 01; 72 h: t = 50. 20, 28. 84and 15. 83, respectively, each P < 0. 01; 96 h: t = 11. 18, 18. 35 and 20. 92, respectively, each P < 0. 01) and clone formation(t = 4. 618, 9. 821 and 12. 14, respectively, each P < 0. 05) of 2. 5, 5 and 10 μmol/L dose groups were significantly lower than those of the control group. Compared with the control group, the proportion of cells in S phase of three test group increased in a concentration-dependent manner(48 h: t = 6. 316, 8. 156 and 14. 11, respectively, each P < 0. 05; 72 h: t =7. 231, 15. 36 and 25. 16, respectively, each P < 0. 05), and the expression of c-Myc(5 and 10 μmol/L:t = 10. 39 and12. 18, P < 0. 05 and < 0. 01, respectively), CDK1(t = 3. 777, 5. 069 and 6. 974, respectively, each P < 0. 05), CDK2(5 and10 μmol/L:t = 12. 72 and 19. 43, respectively, each P < 0. 01) and cyclinE1(t = 3. 813 and 15. 23, respectively, each P <0. 01) was down-regulated at the protein level. Compared with the control group, each test group significantly inhibited wound healing(48 h: t = 6. 969, 56. 16 and 27. 73, respectively, each P < 0. 05; 72 h: t = 8. 619, 22. 12 and 32. 15, respectively, each P < 0. 05), migration(t = 9. 817, 14. 74 and 19. 39, respectively, each P < 0. 01) and invasion(t = 5. 614, 13. 85and 14. 22, respectively, each P < 0. 01) of MDA-MB-231 cells, and up-regulated the expression of E-cadherin(10 μmol/L:t = 11. 79, P < 0. 01), down-regulated the expression of Vimentin(5 and 10 μmol/L:t = 12. 05 and 13. 02, respectively,each P < 0. 01) and β-catenin(5 and 10 μmol/L:t = 4. 516 and 9. 305, respectively, each P < 0. 01) at the protein level. In addition, the test groups of Cynanoside H could significantly down-regulate the protein expression of PDGFRB(5 and10 μmol/L:t = 7. 083 and 18. 87, respectively, each P < 0. 01) and inhibit the protein levels of p-JAK2(t = 4. 050, 10. 95and 11. 05, respectively, each P < 0. 05) and p-STAT3(5 and 10 μmol/L:t = 15. 25 and 25. 89, respectively, each P <0. 01), but had no effect on the total protein levels of JAK2 and STAT3.Conclusion Cynanoside H inhibits the proliferation of breast cancer MDA-MB-231 cells by inducing S-phase cell cycle arrest and reverses the epithelial-mesenchymal transition(EMT) to inhibit cell metastasis. These effects may be mediated by down-regulating PDGFRB/JAK2/STAT3 signaling pathway.
