Molecular mechanism of anticancer effect of Sclerotium rolfsii lectin in HT29 cells involves differential expression of genes associated with multiple signaling pathways: A microarray analysis.
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.
Department of Studies in Biochemistry, Karnatak University, Dharwad 580 003, India.
Srikanth
S
Barkeer
Agilent Technologies India Pvt. Ltd, Bangalore 560048, India.
Nilanjan
N
Guha
Department of Studies in Biochemistry, Karnatak University, Dharwad 580 003, India.
Vishwanathreddy
V
Hothpet
Agilent Technologies India Pvt. Ltd, Bangalore 560048, India.
Deepak
D
Saligrama Adavigowda
Department of Studies in Biochemistry, Karnatak University, Dharwad 580 003, India.
Prajna
P
Hegde
Agilent Technologies India Pvt. Ltd, Bangalore 560048, India.
Arunkumar
A
Padmanaban
Department of Gastroenterology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, UK.
Lu-Gang
LG
Yu
Department of Studies in Biochemistry, Karnatak University, Dharwad 580 003, India Department of Gastroenterology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, UK.
Bale M
BM
Swamy
Department of Studies in Biochemistry, Karnatak University, Dharwad 580 003, India Department of Gastroenterology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, UK pinkisri2001@yahoo.co.in srinamdar2009@gmail.com.
Shashikala R
SR
Inamdar
D000363:Agaricales / Q000737:chemistry; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D004261:DNA Replication / Q000187:drug effects; D005656:Fungal Proteins / Q000494:pharmacology*; D006801:Humans; D037102:Lectins / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D059467:Transcriptome
D000970:Antineoplastic Agents; D005656:Fungal Proteins; D037102:Lectins
GeneSpring GX 12.6.1; HT29 cells; Microarray; Sclerotium rolfsii lectin; mRNA and miRNA