Silibinin Inhibits TGF-β-induced MMP-2 and MMP-9 Through Smad Signaling Pathway in Colorectal Cancer HT-29 Cells
-
Zahra Zare
,
Tina Nayerpour Dizaj
,
Armaghan Lohrasbi
,
Zakieh Sadat Sheikhalishahi
,
Amirhooman Asadi
,
Mana Zakeri
,
Fahimeh Hosseinabadi
,
Omid Abazari
,
Mojtaba Abbasi
,
Parisa Khanicheragh
Abstract
Background: Metastasis of cancer cells is the primary responsible for death in patients with colorectal cancer (CRC). Transforming growth factor-β (TGF-β)-induced matrix metalloproteinases (MMPs) are essential for the metastasis process. Silibinin is a natural compound extracted from the Silybum marianum that exhibits anti-neoplastic activity in cancer cell lines. In this study, we evaluated the effects of silibinin on MMP-2 and MMP-9 induced by TGF-β in human HT-29 CRC cell line and the potential mechanism underlying the effects. Methods: The present in vitro study was done on the HT-29 cell line. The HT-29 cell line was cultured in RPMI1640 and exposed to TGF- β (5 ng/ml) in the absence and presence of different concentrations of silibinin (10, 25, 50, and 100 μM). The effect of silibinin on HT-29 cell viability was measured with the MTT assay. A real-time polymerase chain reaction (Real-Time PCR) determined the relative mRNA expression of MMP-2 and MMP-9. Western blotting was employed to examine MMP-2 and MMP 9 protein expression and Smad2 phosphorylation. Results: Silibinin inhibits cell viability of HT-29 cell line at 24 hours in a dose-dependent manner. TGF-β increased the mRNA and protein expression of MMP-2, MMP-9, and phosphorylated Smad2 compared to controls. Pharmacological inhibition with silibinin markedly blocked TGF-β–induced MMP-2 and MMP-9 mRNA and protein expression and Smad2 phosphorylation. Conclusion: Silibinin decreased the cell viability of HT-29 cancer cells in a dose-dependent manner. Silibinin also inhibited TGF-β-stimulated MMP-2 and MMP-9 expression in HT-29 cells, possibly mediated with the Smad2 signaling pathway.
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2. Said AH, Raufman J-P, Xie G. The role of matrix metalloproteinases in colorectal cancer. Cancers. 2014;6(1):366-75.
3. Huang H. Matrix metalloproteinase-9 (MMP-9) as a cancer biomarker and MMP-9 biosensors: recent advances. Sensors. 2018;18(10):3249.
4. Shafaei Z, Abazari O, Divsalar A, Ghalandari B, Poursoleiman A, Saboury AA, et al. Effect of a Synthesized Amyl-Glycine1, 10-Phenanthroline Platinum Nitrate on Structure and Stability of Human Blood Carrier Protein, Albumin: Spectroscopic and Modeling Approaches. Journal of fluorescence. 2017;27(5):1829-38.
5. Bates AL, Pickup MW, Hallett MA, Dozier EA, Thomas S, Fingleton B. Stromal matrix metalloproteinase 2 regulates collagen expression and promotes the outgrowth of experimental metastases. The Journal of pathology. 2015;235(5):773-83.
6. Mo N, Li Z-Q, Li J, Cao Y-D. Curcumin inhibits TGF-β1-induced MMP-9 and invasion through ERK and Smad signaling in breast cancer MDA-MB-231 cells. Asian Pacific Journal of Cancer Prevention. 2012;13(11):5709-14.
7. Kim E-S, Kim M-S, Moon A. TGF-β-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK, but not ERK signaling in MCF10A human breast epithelial cells. International journal of oncology. 2004;25(5):1375-82.
8. Massagué J. TGFβ signalling in context. Nature reviews Molecular cell biology. 2012;13(10):616-30.
9. Mohamed R, Dayati P, Mehr RN, Kamato D, Seif F, Babaahmadi-Rezaei H, et al. Transforming growth factor–β1 mediated CHST11 and CHSY1 mRNA expression is ROS dependent in vascular smooth muscle cells. Journal of cell Communication and Signaling. 2019;13(2):225-33.
10. Hata A, Chen Y-G. TGF-β signaling from receptors to Smads. Cold Spring Harbor perspectives in biology. 2016;8(9):a022061.
11. Dayati P, Rezaei HB, Sharifat N, Kamato D, Little PJ. G protein coupled receptors can transduce signals through carboxy terminal and linker region phosphorylation of Smad transcription factors. Life sciences. 2018;199:10-5.
12. Webb AH, Gao BT, Goldsmith ZK, Irvine AS, Saleh N, Lee RP, et al. Inhibition of MMP-2 and MMP-9 decreases cellular migration, and angiogenesis in in vitro models of retinoblastoma. BMC cancer. 2017;17(1):434.
13. Chen S, Zhu J, Zuo S, Ma J, Zhang J, Chen G, et al. 1, 25 (OH) 2D3 attenuates TGF-β1/β2-induced increased migration and invasion via inhibiting epithelial–mesenchymal transition in colon cancer cells. Biochemical and biophysical research communications. 2015;468(1-2):130-5.
14. Abazari O, Shafaei Z, Divsalar A, Eslami-Moghadam M, Ghalandari B, Saboury AA, et al. Interaction of the synthesized anticancer compound of the methyl-glycine 1, 10-phenanthroline platinum nitrate with human serum albumin and human hemoglobin proteins by spectroscopy methods and molecular docking. Journal of the Iranian Chemical Society. 2020:1-14.
15. Raina K, Kumar S, Dhar D, Agarwal R. Silibinin and colorectal cancer chemoprevention: a comprehensive review on mechanisms and efficacy. Journal of biomedical research. 2016;30(6):452.
16. Sharifat N, Jafari-Hafshejani F, Dayati P, Lorestanpoor P, Paydar A, BabaahmadiRezaei H. Inhibitory effect of Curcumin on phosphorylation NFκB-p65 induced by hydrogen peroxide in Bovine Endothelial Cells. Journal of Fasa University of Medical Sciences. 2017;7(2):283-90.
17. Wing Ying Cheung C, Gibbons N, Wayne Johnson D, Lawrence Nicol D. Silibinin-a promising new treatment for cancer. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents). 2010;10(3):186-95.
18. Kim S, Kim SH, Hur SM, Lee S-K, Kim WW, Kim JS, et al. Silibinin prevents TPA-induced MMP-9 expression by down-regulation of COX-2 in human breast cancer cells. Journal of ethnopharmacology. 2009;126(2):252-7.
19. Imai-Sumida M, Chiyomaru T, Majid S, Saini S, Nip H, Dahiya R, et al. Silibinin suppresses bladder cancer through down-regulation of actin cytoskeleton and PI3K/Akt signaling pathways. Oncotarget. 2017;8(54):92032.
20. Prasad RR, Paudel S, Raina K, Agarwal R. Silibinin and non-melanoma skin cancers. Journal of Traditional and Complementary Medicine. 2020.
21. Zhu X-X, Ding Y-H, Wu Y, Qian L-Y, Zou H, He Q. Silibinin: a potential old drug for cancer therapy. Expert review of clinical pharmacology. 2016;9(10):1323-30.
22. Abazari O, Divsalar A, Ghobadi R. Inhibitory effects of oxali-Platin as a chemotherapeutic drug on the function and structure of bovine liver catalase. Journal of Biomolecular Structure and Dynamics. 2019:1-7.
23. Lin C-M, Chen Y-H, Ma H-P, Wang B-W, Chiu J-H, Chua S-K, et al. Silibinin inhibits the invasion of IL-6-stimulated colon cancer cells via selective JNK/AP-1/MMP-2 modulation in vitro. Journal of agricultural and food chemistry. 2012;60(51):12451-7.
24. Deep G, Agarwal R. Antimetastatic efficacy of silibinin: molecular mechanisms and therapeutic potential against cancer. Cancer and Metastasis Reviews. 2010;29(3):447-63.
25. Bai X, Li Y-y, Zhang H-y, Wang F, He H-l, Yao J-c, et al. Role of matrix metalloproteinase-9 in transforming growth factor-β1-induced epithelial–mesenchymal transition in esophageal squamous cell carcinoma. OncoTargets and therapy. 2017;10:2837.
26. Moore-Smith LD, Isayeva T, Lee JH, Frost A, Ponnazhagan S. Silencing of TGF-β1 in tumor cells impacts MMP-9 in tumor microenvironment. Scientific Reports. 2017;7(1):1-10.
27. Eble JA, Niland S. The extracellular matrix in tumor progression and metastasis. Clinical & experimental metastasis. 2019:1-28.
28. Abazari O, Shafaei Z, Divsalar A, Eslami-Moghadam M, Ghalandari B, Saboury AA. Probing the biological evaluations of a new designed Pt (II) complex using spectroscopic and theoretical approaches: Human hemoglobin as a target. Journal of Biomolecular Structure and Dynamics. 2016;34(5):1123-31.
29. Babykutty S, PS P, RJ N, Kumar MS, Nair MS, Srinivas P, et al. Nimbolide retards tumor cell migration, invasion, and angiogenesis by downregulating MMP‐2/9 expression via inhibiting ERK1/2 and reducing DNA‐binding activity of NF‐κB in colon cancer cells. Molecular carcinogenesis. 2012;51(6):475-90.
30. Abazari O, Divsalar A, Ghobadi R. Inhibitory effects of oxali-Platin as a chemotherapeutic drug on the function and structure of bovine liver catalase. Journal of Biomolecular Structure and Dynamics. 2020;38(2):609-15.
31. Mo N, Li Z-Q, Li J, Cao Y-D. Curcumin inhibits TGF-β1-induced MMP-9 and invasion through ERK and Smad signaling in breast cancer MDA-MB-231 cells. Asian Pac J Cancer Prev. 2012;13(11):5709-14.
32. Kim S, Han J, Jeon M, You D, Lee J, Kim HJ, et al. Silibinin inhibits triple negative breast cancer cell motility by suppressing TGF-β2 expression. Tumor Biology. 2016;37(8):11397-407.
33. Lu S, Zhang Z, Chen M, Li C, Liu L, Li Y. Silibinin inhibits the migration and invasion of human gastric cancer SGC7901 cells by downregulating MMP‑2 and MMP‑9 expression via the p38MAPK signaling pathway. Oncology letters. 2017;14(6):7577-82.
34. Polachi N, Bai G, Li T, Chu Y, Wang X, Li S, et al. Modulatory effects of silibinin in various cell signaling pathways against liver disorders and cancer–A comprehensive review. European journal of medicinal chemistry. 2016;123:577-95.
35. Asadi A, Nezhad DY, Javazm AR, Khanicheragh P, Mashouri L, Shakeri F, et al. In vitro Effects of Curcumin on Transforming Growth Factor-β-mediated Non-Smad Signaling Pathway, Oxidative Stress, and Pro‐inflammatory Cytokines Production with Human Vascular Smooth Muscle Cells. Iranian Journal of Allergy, Asthma and Immunology. 2019:1-10.
2. Said AH, Raufman J-P, Xie G. The role of matrix metalloproteinases in colorectal cancer. Cancers. 2014;6(1):366-75.
3. Huang H. Matrix metalloproteinase-9 (MMP-9) as a cancer biomarker and MMP-9 biosensors: recent advances. Sensors. 2018;18(10):3249.
4. Shafaei Z, Abazari O, Divsalar A, Ghalandari B, Poursoleiman A, Saboury AA, et al. Effect of a Synthesized Amyl-Glycine1, 10-Phenanthroline Platinum Nitrate on Structure and Stability of Human Blood Carrier Protein, Albumin: Spectroscopic and Modeling Approaches. Journal of fluorescence. 2017;27(5):1829-38.
5. Bates AL, Pickup MW, Hallett MA, Dozier EA, Thomas S, Fingleton B. Stromal matrix metalloproteinase 2 regulates collagen expression and promotes the outgrowth of experimental metastases. The Journal of pathology. 2015;235(5):773-83.
6. Mo N, Li Z-Q, Li J, Cao Y-D. Curcumin inhibits TGF-β1-induced MMP-9 and invasion through ERK and Smad signaling in breast cancer MDA-MB-231 cells. Asian Pacific Journal of Cancer Prevention. 2012;13(11):5709-14.
7. Kim E-S, Kim M-S, Moon A. TGF-β-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK, but not ERK signaling in MCF10A human breast epithelial cells. International journal of oncology. 2004;25(5):1375-82.
8. Massagué J. TGFβ signalling in context. Nature reviews Molecular cell biology. 2012;13(10):616-30.
9. Mohamed R, Dayati P, Mehr RN, Kamato D, Seif F, Babaahmadi-Rezaei H, et al. Transforming growth factor–β1 mediated CHST11 and CHSY1 mRNA expression is ROS dependent in vascular smooth muscle cells. Journal of cell Communication and Signaling. 2019;13(2):225-33.
10. Hata A, Chen Y-G. TGF-β signaling from receptors to Smads. Cold Spring Harbor perspectives in biology. 2016;8(9):a022061.
11. Dayati P, Rezaei HB, Sharifat N, Kamato D, Little PJ. G protein coupled receptors can transduce signals through carboxy terminal and linker region phosphorylation of Smad transcription factors. Life sciences. 2018;199:10-5.
12. Webb AH, Gao BT, Goldsmith ZK, Irvine AS, Saleh N, Lee RP, et al. Inhibition of MMP-2 and MMP-9 decreases cellular migration, and angiogenesis in in vitro models of retinoblastoma. BMC cancer. 2017;17(1):434.
13. Chen S, Zhu J, Zuo S, Ma J, Zhang J, Chen G, et al. 1, 25 (OH) 2D3 attenuates TGF-β1/β2-induced increased migration and invasion via inhibiting epithelial–mesenchymal transition in colon cancer cells. Biochemical and biophysical research communications. 2015;468(1-2):130-5.
14. Abazari O, Shafaei Z, Divsalar A, Eslami-Moghadam M, Ghalandari B, Saboury AA, et al. Interaction of the synthesized anticancer compound of the methyl-glycine 1, 10-phenanthroline platinum nitrate with human serum albumin and human hemoglobin proteins by spectroscopy methods and molecular docking. Journal of the Iranian Chemical Society. 2020:1-14.
15. Raina K, Kumar S, Dhar D, Agarwal R. Silibinin and colorectal cancer chemoprevention: a comprehensive review on mechanisms and efficacy. Journal of biomedical research. 2016;30(6):452.
16. Sharifat N, Jafari-Hafshejani F, Dayati P, Lorestanpoor P, Paydar A, BabaahmadiRezaei H. Inhibitory effect of Curcumin on phosphorylation NFκB-p65 induced by hydrogen peroxide in Bovine Endothelial Cells. Journal of Fasa University of Medical Sciences. 2017;7(2):283-90.
17. Wing Ying Cheung C, Gibbons N, Wayne Johnson D, Lawrence Nicol D. Silibinin-a promising new treatment for cancer. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents). 2010;10(3):186-95.
18. Kim S, Kim SH, Hur SM, Lee S-K, Kim WW, Kim JS, et al. Silibinin prevents TPA-induced MMP-9 expression by down-regulation of COX-2 in human breast cancer cells. Journal of ethnopharmacology. 2009;126(2):252-7.
19. Imai-Sumida M, Chiyomaru T, Majid S, Saini S, Nip H, Dahiya R, et al. Silibinin suppresses bladder cancer through down-regulation of actin cytoskeleton and PI3K/Akt signaling pathways. Oncotarget. 2017;8(54):92032.
20. Prasad RR, Paudel S, Raina K, Agarwal R. Silibinin and non-melanoma skin cancers. Journal of Traditional and Complementary Medicine. 2020.
21. Zhu X-X, Ding Y-H, Wu Y, Qian L-Y, Zou H, He Q. Silibinin: a potential old drug for cancer therapy. Expert review of clinical pharmacology. 2016;9(10):1323-30.
22. Abazari O, Divsalar A, Ghobadi R. Inhibitory effects of oxali-Platin as a chemotherapeutic drug on the function and structure of bovine liver catalase. Journal of Biomolecular Structure and Dynamics. 2019:1-7.
23. Lin C-M, Chen Y-H, Ma H-P, Wang B-W, Chiu J-H, Chua S-K, et al. Silibinin inhibits the invasion of IL-6-stimulated colon cancer cells via selective JNK/AP-1/MMP-2 modulation in vitro. Journal of agricultural and food chemistry. 2012;60(51):12451-7.
24. Deep G, Agarwal R. Antimetastatic efficacy of silibinin: molecular mechanisms and therapeutic potential against cancer. Cancer and Metastasis Reviews. 2010;29(3):447-63.
25. Bai X, Li Y-y, Zhang H-y, Wang F, He H-l, Yao J-c, et al. Role of matrix metalloproteinase-9 in transforming growth factor-β1-induced epithelial–mesenchymal transition in esophageal squamous cell carcinoma. OncoTargets and therapy. 2017;10:2837.
26. Moore-Smith LD, Isayeva T, Lee JH, Frost A, Ponnazhagan S. Silencing of TGF-β1 in tumor cells impacts MMP-9 in tumor microenvironment. Scientific Reports. 2017;7(1):1-10.
27. Eble JA, Niland S. The extracellular matrix in tumor progression and metastasis. Clinical & experimental metastasis. 2019:1-28.
28. Abazari O, Shafaei Z, Divsalar A, Eslami-Moghadam M, Ghalandari B, Saboury AA. Probing the biological evaluations of a new designed Pt (II) complex using spectroscopic and theoretical approaches: Human hemoglobin as a target. Journal of Biomolecular Structure and Dynamics. 2016;34(5):1123-31.
29. Babykutty S, PS P, RJ N, Kumar MS, Nair MS, Srinivas P, et al. Nimbolide retards tumor cell migration, invasion, and angiogenesis by downregulating MMP‐2/9 expression via inhibiting ERK1/2 and reducing DNA‐binding activity of NF‐κB in colon cancer cells. Molecular carcinogenesis. 2012;51(6):475-90.
30. Abazari O, Divsalar A, Ghobadi R. Inhibitory effects of oxali-Platin as a chemotherapeutic drug on the function and structure of bovine liver catalase. Journal of Biomolecular Structure and Dynamics. 2020;38(2):609-15.
31. Mo N, Li Z-Q, Li J, Cao Y-D. Curcumin inhibits TGF-β1-induced MMP-9 and invasion through ERK and Smad signaling in breast cancer MDA-MB-231 cells. Asian Pac J Cancer Prev. 2012;13(11):5709-14.
32. Kim S, Han J, Jeon M, You D, Lee J, Kim HJ, et al. Silibinin inhibits triple negative breast cancer cell motility by suppressing TGF-β2 expression. Tumor Biology. 2016;37(8):11397-407.
33. Lu S, Zhang Z, Chen M, Li C, Liu L, Li Y. Silibinin inhibits the migration and invasion of human gastric cancer SGC7901 cells by downregulating MMP‑2 and MMP‑9 expression via the p38MAPK signaling pathway. Oncology letters. 2017;14(6):7577-82.
34. Polachi N, Bai G, Li T, Chu Y, Wang X, Li S, et al. Modulatory effects of silibinin in various cell signaling pathways against liver disorders and cancer–A comprehensive review. European journal of medicinal chemistry. 2016;123:577-95.
35. Asadi A, Nezhad DY, Javazm AR, Khanicheragh P, Mashouri L, Shakeri F, et al. In vitro Effects of Curcumin on Transforming Growth Factor-β-mediated Non-Smad Signaling Pathway, Oxidative Stress, and Pro‐inflammatory Cytokines Production with Human Vascular Smooth Muscle Cells. Iranian Journal of Allergy, Asthma and Immunology. 2019:1-10.
| Files | ||
| Issue | Vol 12 No 2 (2020) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/bccr.v12i2.5752 | |
| Keywords | ||
| Silibinin Colorectal Neoplasms TGF-β MMP-2 MMP-9 | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Zare Z, Nayerpour Dizaj T, Lohrasbi A, Sheikhalishahi ZS, Asadi A, Zakeri M, Hosseinabadi F, Abazari O, Abbasi M, Khanicheragh P. Silibinin Inhibits TGF-β-induced MMP-2 and MMP-9 Through Smad Signaling Pathway in Colorectal Cancer HT-29 Cells. Basic Clin Cancer Res. 2021;12(2):81-90.
