With specific designed prmers. CYP2B6 and CYP1A1 cDNA were generatecl by reverse transcrlI7tion-Polymerase chain reaction(RT-PCR )technlque Performed on total RNAs isolated frorn hum1ln liver and 3-rnethylch(,lanthren...With specific designed prmers. CYP2B6 and CYP1A1 cDNA were generatecl by reverse transcrlI7tion-Polymerase chain reaction(RT-PCR )technlque Performed on total RNAs isolated frorn hum1ln liver and 3-rnethylch(,lanthrene(3-Mtt)induc human amnion FL, cells. Cell llnes (CHL, 2B6 and CtHL-1A1 ) capableof expressing hunlan cytochome P 15O (CYP ) 2B6 and 1A1 were establishecl after transfection of corre-sponding eukaryotic reconlbinant expression plasmid with human CYP2ll6 and 1A1 cDNA lnserts respectlvely. These cell lines stably expressed the mRNAs and the enzymatic activltles cc)rresI’onding to ttYP2B6and CYP1A1, respectively’ Compared with Chinese hamster lung (CHL) cells, the n1icr()nucleus frecluencyin CHl,-2B6 cells is markedly lncreased when exPosed to nitrosamines,aflatoxln B, (AFB1) and cyclophos-Phamide (CPA). Thls is also in CHL-1A1 cells,when exposed to carcinogenic polycycllc aromatic hydrocar-bons.展开更多
Objective： Fruit of Phyllanthus emblica Linn. （PE） is widely consumed as a functional food and used as a folk medicine due to its remarkable nutritional and pharmacological effects. Mitomycin C （MMC） and cisplat...Objective： Fruit of Phyllanthus emblica Linn. （PE） is widely consumed as a functional food and used as a folk medicine due to its remarkable nutritional and pharmacological effects. Mitomycin C （MMC） and cisplatin （cDDP） are the most widely used forms of chemotherapeutic drug, but their clinical use is limited by their genotoxicity to normal cells. We aimed to determine whether PE has potential to reduce the genotoxicity, while improving the anticancer effect, of MMC and cDDP. Methods： Cell proliferation was evaluated using the trypan blue exclusion assay and colony-forming assay. Genomic instability （GIN） was measured using the cytokinesis-block micronucieus assay. Results： Co-treatment （72 h） with PE at 20-320 μg/ml significantly enhanced the efficacy of MMC （0.05 μg/ml） and cDDP （1 μg/ml） against Colo205 colorectal cancer cells （P〈0.05）, and at 80-320 μg/ml significantly decreased MMC- and cDDP-induced GIN and multinucleation in normal colonic NCM460 cells （P〈0.05）. PE significantly decreased the mitotic index （P〈0.01）, blocked mitotic progression （P〈0.05）, and promoted apoptosis （P〈0.01） in MMC- and cDDP-treated NCM460 cells, suggesting that PE-mediated inhibition of mitosis and induction of apoptosis may limit the division and survival of highly damaged cells. Also, PE was found to inhibit the clonal expansion of MMC- and cDDP-treated NCM460 cells （P〈0.05） and decrease the heterogeneity of the surviving clones. Conclusions： PE potentiates the anticancer efficacy of MMC and cDDP, while preventing their genotoxicity and inhibiting clonal expansions of unstable genomes in normal cells. These data suggest that PE has the potential to reduce the risk of secondary cancers induced by chemotherapeutics.展开更多
文摘With specific designed prmers. CYP2B6 and CYP1A1 cDNA were generatecl by reverse transcrlI7tion-Polymerase chain reaction(RT-PCR )technlque Performed on total RNAs isolated frorn hum1ln liver and 3-rnethylch(,lanthrene(3-Mtt)induc human amnion FL, cells. Cell llnes (CHL, 2B6 and CtHL-1A1 ) capableof expressing hunlan cytochome P 15O (CYP ) 2B6 and 1A1 were establishecl after transfection of corre-sponding eukaryotic reconlbinant expression plasmid with human CYP2ll6 and 1A1 cDNA lnserts respectlvely. These cell lines stably expressed the mRNAs and the enzymatic activltles cc)rresI’onding to ttYP2B6and CYP1A1, respectively’ Compared with Chinese hamster lung (CHL) cells, the n1icr()nucleus frecluencyin CHl,-2B6 cells is markedly lncreased when exPosed to nitrosamines,aflatoxln B, (AFB1) and cyclophos-Phamide (CPA). Thls is also in CHL-1A1 cells,when exposed to carcinogenic polycycllc aromatic hydrocar-bons.
基金Project supported by the National Natural Science Foundation of China(Nos.31260268 and 31560307)
文摘Objective： Fruit of Phyllanthus emblica Linn. （PE） is widely consumed as a functional food and used as a folk medicine due to its remarkable nutritional and pharmacological effects. Mitomycin C （MMC） and cisplatin （cDDP） are the most widely used forms of chemotherapeutic drug, but their clinical use is limited by their genotoxicity to normal cells. We aimed to determine whether PE has potential to reduce the genotoxicity, while improving the anticancer effect, of MMC and cDDP. Methods： Cell proliferation was evaluated using the trypan blue exclusion assay and colony-forming assay. Genomic instability （GIN） was measured using the cytokinesis-block micronucieus assay. Results： Co-treatment （72 h） with PE at 20-320 μg/ml significantly enhanced the efficacy of MMC （0.05 μg/ml） and cDDP （1 μg/ml） against Colo205 colorectal cancer cells （P〈0.05）, and at 80-320 μg/ml significantly decreased MMC- and cDDP-induced GIN and multinucleation in normal colonic NCM460 cells （P〈0.05）. PE significantly decreased the mitotic index （P〈0.01）, blocked mitotic progression （P〈0.05）, and promoted apoptosis （P〈0.01） in MMC- and cDDP-treated NCM460 cells, suggesting that PE-mediated inhibition of mitosis and induction of apoptosis may limit the division and survival of highly damaged cells. Also, PE was found to inhibit the clonal expansion of MMC- and cDDP-treated NCM460 cells （P〈0.05） and decrease the heterogeneity of the surviving clones. Conclusions： PE potentiates the anticancer efficacy of MMC and cDDP, while preventing their genotoxicity and inhibiting clonal expansions of unstable genomes in normal cells. These data suggest that PE has the potential to reduce the risk of secondary cancers induced by chemotherapeutics.
