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Published on: January 22, 2021
Molecular mechanism of anticancer effect of Sclerotium rolfsii lectin in HT29 cells involves differential expression
Srikanth Barkeer1, Nilanjan Guha2, Vishwanathreddy Hothpet1
1Department of Studies in Biochemistry, Karnatak University, Dharwad 580 003, India.
Abstract:
Sclerotium rolfsii lectin (SRL) is a lectin isolated from fungus S. rolfsii and has high binding specificity toward the oncofetal Thomsen-Friedenreich carbohydrate antigen (Galβ1-3GalNAc-α-O-Ser/Thr, T or TF), which is expressed in more than 90% of human cancers. Our previous studies have shown that binding of SRL to human colon, breast and ovarian cancer cells induces cell apoptosis in vitro and suppresses tumor growth in vivo. This study investigated the SRL-mediated cell signaling in human colon cancer HT29 cells by mRNA and miRNA microarrays. It was found that SRL treatment results in altered expression of several hundred molecules including mitogen-activated protein kinase (MAPK) and c-JUN-associated, apoptosis-associated and cell cycle and DNA replication-associated signaling molecules. Pathway analysis using GeneSpring 12.6.1 revealed that SRL treatment induces changes of MAPK and c-JUN-associated signaling pathways as early as 2 h while changes of cell cycle, DNA replication and apoptosis pathways were significantly affected only after 24 h. A significant change of cell miRNA expression was also observed after 12 h treatment of the cells with SRL. These changes were further validated by quantitative real time polymerase chain reaction and immunoblotting. This study thus suggests that the presence of SRL affects multiple signaling pathways in cancer cells with early effects on cell proliferation pathways associated with MAPK and c-JUN, followed by miRNA-associated cell activity and apoptosis. This provides insight information into the molecular mechanism of the anticancer activity of this fungal lectin.
Insights
Sclerotium rolfsii lectin (SRL) impacts cancer cell signaling. Early effects on MAPK and c-JUN pathways precede changes in miRNA expression and apoptosis, revealing the fungal lectin's anticancer mechanisms.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Sclerotium rolfsii lectin (SRL) specifically binds the Thomsen-Friedenreich antigen (T or TF), prevalent in human cancers.
- Previous research demonstrated SRL's ability to induce apoptosis in cancer cells and suppress tumor growth.
Purpose of the Study:
- To investigate the molecular mechanisms of SRL-mediated cell signaling in human colon cancer HT29 cells.
- To identify key signaling pathways and molecular changes induced by SRL treatment.
Main Methods:
- Utilized mRNA and miRNA microarrays to analyze gene and microRNA expression profiles.
- Performed pathway analysis, quantitative real-time polymerase chain reaction, and immunoblotting for validation.
Main Results:
- SRL treatment altered hundreds of molecules, including those in MAPK, c-JUN, apoptosis, cell cycle, and DNA replication pathways.
- Early changes (2h) were observed in MAPK and c-JUN signaling, followed by cell cycle/DNA replication pathway alterations (24h) and miRNA expression changes (12h).
Conclusions:
- SRL affects multiple signaling pathways in cancer cells, initiating with proliferation pathways (MAPK, c-JUN).
- Subsequent effects involve miRNA modulation and induction of apoptosis, providing insight into SRL's anticancer activity.
