Co-overexpression of geraniol-10-hydroxylase and strictosidine synthase improves anti-cancer drug camptothecin accumulation in Ophiorrhiza pumila.

Abstract

Camptothecin (CPT) belongs to a group of monoterpenoidindole alkaloids (TIAs) and its derivatives such as irinothecan and topothecan have been widely used worldwide for the treatment of cancer, giving rise to rapidly increasing market demands. Genes from Catharanthus roseus encoding strictosidine synthase (STR) and geraniol 10-hydroxylase (G10H), were separately and simultaneously introduced into Ophiorrhiza pumila hairy roots. Overexpression of individual G10H (G lines) significantly improved CPT production with respect to non-transgenic hairy root cultures (NC line) and single STR overexpressing lines (S lines), indicating that G10H plays a more important role in stimulating CPT accumulation than STR in O. pumila. Furthermore, co-overexpression of G10H and STR genes (SG Lines) caused a 56% increase on the yields of CPT compared to NC line and single gene transgenic lines, showed that simultaneous introduction of G10H and STR can produce a synergistic effect on CPT biosynthesis in O. pumila. The MTT assay results indicated that CPT extracted from different lines showed similar anti-tumor activity, suggesting that transgenic O. pumila hairy root lines could be an alternative approach to obtain CPT. To our knowledge, this is the first report on the enhancement of CPT production in O. pumila employing a metabolic engineering strategy.

• Authors: Lijie Cui, Xiaoling Ni, Qian Ji, Xiaojuan Teng, Yanru Yang, Chao Wu, David Zekria, Dasheng Zhang, Guoyin Kai
• Journal: Scientific reports (Sci Rep) Vol. 5 Pg. 8227 (Feb 04 2015) ISSN: 2045-2322 [Electronic] England
• PMID: 25648209 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents, Phytogenic
• Cytochrome P-450 Enzyme System
• geraniol 10-hydroxylase
• Carbon-Nitrogen Lyases
• strictosidine synthetase
• Camptothecin

Topics

• Antineoplastic Agents, Phytogenic (biosynthesis, metabolism, pharmacology)
• Biosynthetic Pathways
• Camptothecin (biosynthesis, metabolism, pharmacology)
• Carbon-Nitrogen Lyases (genetics, metabolism)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Cytochrome P-450 Enzyme System (genetics, metabolism)
• Gene Expression
• Humans
• Leukemia, Myeloid
• Plant Roots (genetics)
• Plants, Genetically Modified
• Rubiaceae (genetics, metabolism)
• Transcription, Genetic