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Inhibition of cancer cell proliferation and apoptosis-inducing activity of fungal taxol and its precursor baccatin III purified from endophytic Fusarium solani.

Chakravarthi BV, Sujay R, Kuriakose GC, Karande AA, Jayabaskaran C - Cancer Cell Int. (2013)

Bottom Line: They also induced apoptosis in JR4-Jurkat cells with a possible involvement of anti-apoptotic Bcl2 and loss in mitochondrial membrane potential, and was unaffected by inhibitors of caspase-9,-2 or -3 but was prevented in presence of caspase-10 inhibitor.DNA fragmentation was also observed in cells treated with fungal taxol and baccatin III.The cytotoxic activity exhibited by fungal taxol and baccatin III involves the same mechanism, dependent on caspase-10 and membrane potential loss of mitochondria, with taxol having far greater cytotoxic potential.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India. cjb@biochem.iisc.ernet.in.

ABSTRACT

Background: Taxol (generic name paclitaxel), a plant-derived antineoplastic agent, used widely against breast, ovarian and lung cancer, was originally isolated from the bark of the Pacific yew, Taxus brevifolia. The limited supply of the drug has prompted efforts to find alternative sources, such as chemical synthesis, tissue and cell cultures of the Taxus species both of which are expensive and yield low levels. Fermentation processes with microorganisms would be the methods of choice to lower the costs and increase yields. Previously we have reported that F. solani isolated from T. celebica produced taxol and its precursor baccatin III in liquid grown cultures J Biosci 33:259-67, 2008. This study was performed to evaluate the inhibition of proliferation and induction of apoptosis of cancer cell lines by the fungal taxol and fungal baccatin III of F. solani isolated from T. celebica.

Methods: Cell lines such as HeLa, HepG2, Jurkat, Ovcar3 and T47D were cultured individually and treated with fungal taxol, baccatin III with or without caspase inhibitors according to experimental requirements. Their efficacy on apoptotic induction was examined.

Results: Both fungal taxol and baccatin III inhibited cell proliferation of a number of cancer cell lines with IC50 ranging from 0.005 to 0.2 μM for fungal taxol and 2 to 5 μM for fungal baccatin III. They also induced apoptosis in JR4-Jurkat cells with a possible involvement of anti-apoptotic Bcl2 and loss in mitochondrial membrane potential, and was unaffected by inhibitors of caspase-9,-2 or -3 but was prevented in presence of caspase-10 inhibitor. DNA fragmentation was also observed in cells treated with fungal taxol and baccatin III.

Conclusions: The cytotoxic activity exhibited by fungal taxol and baccatin III involves the same mechanism, dependent on caspase-10 and membrane potential loss of mitochondria, with taxol having far greater cytotoxic potential.

No MeSH data available.


Related in: MedlinePlus

DNA fragmentation of JR4-Jurkat and HeLa cells induced upon treatment with fungal taxol and baccatin III. (A) JR4-Jurkat and (B) HeLa cells (3 × 106) were treated independently with fungal taxol, baccatin III or DMSO at indicated concentrations for 36 h. After extraction, DNA was electrophoresed in a 1.2% agarose gel, stained with ethidium bromide and photographed under UV illumination.
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Figure 7: DNA fragmentation of JR4-Jurkat and HeLa cells induced upon treatment with fungal taxol and baccatin III. (A) JR4-Jurkat and (B) HeLa cells (3 × 106) were treated independently with fungal taxol, baccatin III or DMSO at indicated concentrations for 36 h. After extraction, DNA was electrophoresed in a 1.2% agarose gel, stained with ethidium bromide and photographed under UV illumination.

Mentions: The fragmentation of nuclear DNA is one of the hallmarks of apoptosis. It is known that DNA fragmentation is carried out by the caspase activated DNase (CAD). Activation of CAD leads to cleavage of nuclear DNA into multiples of ~200 bp oligonucleosomal size fragments. To confirm the induction of apoptosis, JR4-Jurkat and HeLa cells were treated with fungal taxol or baccatin III. Low molecular weight DNA isolated from these cells was analyzed in 1.2% agarose gels. DNA ladder formation is observed upon taxol or baccatin III treatment in JR4-Jurkat and HeLa cells, while there is no DNA fragmentation seen in untreated and DMSO treated cells (Figure 7). This confirmed that both fungal taxol and baccatin III can induce apoptosis in JR4-Jurkat or HeLa cells.


Inhibition of cancer cell proliferation and apoptosis-inducing activity of fungal taxol and its precursor baccatin III purified from endophytic Fusarium solani.

Chakravarthi BV, Sujay R, Kuriakose GC, Karande AA, Jayabaskaran C - Cancer Cell Int. (2013)

DNA fragmentation of JR4-Jurkat and HeLa cells induced upon treatment with fungal taxol and baccatin III. (A) JR4-Jurkat and (B) HeLa cells (3 × 106) were treated independently with fungal taxol, baccatin III or DMSO at indicated concentrations for 36 h. After extraction, DNA was electrophoresed in a 1.2% agarose gel, stained with ethidium bromide and photographed under UV illumination.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016216&req=5

Figure 7: DNA fragmentation of JR4-Jurkat and HeLa cells induced upon treatment with fungal taxol and baccatin III. (A) JR4-Jurkat and (B) HeLa cells (3 × 106) were treated independently with fungal taxol, baccatin III or DMSO at indicated concentrations for 36 h. After extraction, DNA was electrophoresed in a 1.2% agarose gel, stained with ethidium bromide and photographed under UV illumination.
Mentions: The fragmentation of nuclear DNA is one of the hallmarks of apoptosis. It is known that DNA fragmentation is carried out by the caspase activated DNase (CAD). Activation of CAD leads to cleavage of nuclear DNA into multiples of ~200 bp oligonucleosomal size fragments. To confirm the induction of apoptosis, JR4-Jurkat and HeLa cells were treated with fungal taxol or baccatin III. Low molecular weight DNA isolated from these cells was analyzed in 1.2% agarose gels. DNA ladder formation is observed upon taxol or baccatin III treatment in JR4-Jurkat and HeLa cells, while there is no DNA fragmentation seen in untreated and DMSO treated cells (Figure 7). This confirmed that both fungal taxol and baccatin III can induce apoptosis in JR4-Jurkat or HeLa cells.

Bottom Line: They also induced apoptosis in JR4-Jurkat cells with a possible involvement of anti-apoptotic Bcl2 and loss in mitochondrial membrane potential, and was unaffected by inhibitors of caspase-9,-2 or -3 but was prevented in presence of caspase-10 inhibitor.DNA fragmentation was also observed in cells treated with fungal taxol and baccatin III.The cytotoxic activity exhibited by fungal taxol and baccatin III involves the same mechanism, dependent on caspase-10 and membrane potential loss of mitochondria, with taxol having far greater cytotoxic potential.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India. cjb@biochem.iisc.ernet.in.

ABSTRACT

Background: Taxol (generic name paclitaxel), a plant-derived antineoplastic agent, used widely against breast, ovarian and lung cancer, was originally isolated from the bark of the Pacific yew, Taxus brevifolia. The limited supply of the drug has prompted efforts to find alternative sources, such as chemical synthesis, tissue and cell cultures of the Taxus species both of which are expensive and yield low levels. Fermentation processes with microorganisms would be the methods of choice to lower the costs and increase yields. Previously we have reported that F. solani isolated from T. celebica produced taxol and its precursor baccatin III in liquid grown cultures J Biosci 33:259-67, 2008. This study was performed to evaluate the inhibition of proliferation and induction of apoptosis of cancer cell lines by the fungal taxol and fungal baccatin III of F. solani isolated from T. celebica.

Methods: Cell lines such as HeLa, HepG2, Jurkat, Ovcar3 and T47D were cultured individually and treated with fungal taxol, baccatin III with or without caspase inhibitors according to experimental requirements. Their efficacy on apoptotic induction was examined.

Results: Both fungal taxol and baccatin III inhibited cell proliferation of a number of cancer cell lines with IC50 ranging from 0.005 to 0.2 μM for fungal taxol and 2 to 5 μM for fungal baccatin III. They also induced apoptosis in JR4-Jurkat cells with a possible involvement of anti-apoptotic Bcl2 and loss in mitochondrial membrane potential, and was unaffected by inhibitors of caspase-9,-2 or -3 but was prevented in presence of caspase-10 inhibitor. DNA fragmentation was also observed in cells treated with fungal taxol and baccatin III.

Conclusions: The cytotoxic activity exhibited by fungal taxol and baccatin III involves the same mechanism, dependent on caspase-10 and membrane potential loss of mitochondria, with taxol having far greater cytotoxic potential.

No MeSH data available.


Related in: MedlinePlus