title: 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: Laboratory of Plant Biotechnology, Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China.
authors: Lijie
authors: L
authors: Cui
authors: Department of General Surgery, Zhongshan Hospital, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors: Xiaoling
authors: X
authors: Ni
authors: Laboratory of Plant Biotechnology, Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China.
authors: Qian
authors: Q
authors: Ji
authors: Laboratory of Plant Biotechnology, Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China.
authors: Xiaojuan
authors: X
authors: Teng
authors: Laboratory of Plant Biotechnology, Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China.
authors: Yanru
authors: Y
authors: Yang
authors: Laboratory of Plant Biotechnology, Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China.
authors: Chao
authors: C
authors: Wu
authors: Department of General Surgery, Zhongshan Hospital, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors: David
authors: D
authors: Zekria
authors: Shanghai Chenshan Plant Science Research Center, Shanghai Chenshan Botanical Garden, Chinese Academy of Sciences, Shanghai201602, China.
authors: Dasheng
authors: D
authors: Zhang
authors: 1] Laboratory of Plant Biotechnology, Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China [2] Zhejiang Provincial Key Laboratory for Genetic Improvement and Quality Control of Medicinal Plants, Hangzhou Normal University, Hangzhou 310018, China.
authors: Guoyin
authors: G
authors: Kai
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology; D053898:Biosynthetic Pathways; D002166:Camptothecin / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology; D019759:Carbon-Nitrogen Lyases / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007951:Leukemia, Myeloid; D018517:Plant Roots / Q000235:genetics; D030821:Plants, Genetically Modified; D019666:Rubiaceae / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D003577:Cytochrome P-450 Enzyme System; C081675:geraniol 10-hydroxylase; D019759:Carbon-Nitrogen Lyases; C022684:strictosidine synthetase; D002166:Camptothecin
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