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Background: A lectin from the plant pathogenic fungus Rhizoctonia bataticola (RBL) has shown unique sugarbinding specificity towards the complex N-glycans, which are known to be expressed in many cancers, including ovarian cancer. Previous studies have demonstrated that RBL exerts cytotoxic effects on human ovarian cancer PA-1 cells. In the present study, we determined the signaling mechanism of RBL-induced cytotoxicity in PA-1 cells upon binding to cell surface receptors and also assessed the anti-tumor activity of RBL in NOD-SCID mice bearing ovarian xenografts. Methods: Cytotoxicity was determined by MTT assay, Cell cycle analysis, Annexin V staining, and expression of caspase-3 was determined by flow cytometry using specific fluorescent detection reagents. Western blotting was carried out to observe the intermediates of apoptotic pathway. Results: Treatment of PA-1 cells with RBL decreased their viability in a dose- and time- dependent manner. Cell cycle analysis revealed an increase in the hypo diploid population in response to lectin treatment. RBL-treated cells stained with Annexin V and PI showed an increase in both the early and late apoptotic populations. The mechanism of RBL-induced apoptosis in PA-1 cells was assessed by the observed decrease in MMP (ΔΨm), release of cytochrome-c, activation of caspase-9 and -3, and cleavage of PARP. Specific pharmacological inhibitors revealed the involvement of caspase-9 and -3 but not caspase-8. RBL at a subtoxic concentration (50 μg/mouse) suppressed tumor growth in NOD-SCID mice bearing PA-1 xenografts. Conclusions: The present study demonstrates that, RBL induced cytotoxicity in PA-1 cells is mediated by intrinsic apoptotic pathway. The present study strengthens the merit of RBL for its potential application in ovarian cancer research.

Rhizoctonia Bataticola Lectin (RBL) Induces Apoptosis in Human Ovarian Cancer PA-1 Cells and Suppresses Tumor Growth In Vivo