Breast cancer resistance protein(BCRP)/ATP-binding cassette subfamily G member 2(ABCG2) is an ATP-binding cassette(ABC) transporter identified as a molecular cause of multidrug resistance(MDR) in diverse cancer cells....Breast cancer resistance protein(BCRP)/ATP-binding cassette subfamily G member 2(ABCG2) is an ATP-binding cassette(ABC) transporter identified as a molecular cause of multidrug resistance(MDR) in diverse cancer cells.BCRP physiologically functions as a part of a self-defense mechanism for the organism;it enhances elimination of toxic xenobiotic substances and harmful agents in the gut and biliary tract,as well as through the blood-brain,placental,and possibly blood-testis barriers.BCRP recognizes and transports numerous anticancer drugs including conventional chemotherapeutic and targeted small therapeutic molecules relatively new in clinical use.Thus,BCRP expression in cancer cells directly causes MDR by active efflux of anticancer drugs.Because BCRP is also known to be a stem cell marker,its expression in cancer cells could be a manifestation of metabolic and signaling pathways that confer multiple mechanisms of drug resistance,self-renewal(stemness),and invasiveness(aggressiveness),and thereby impart a poor prognosis.Therefore,blocking BCRP-mediated active efflux may provide a therapeutic benefit for cancers.Delineating the precise molecular mechanisms for BCRP gene expression may lead to identification of a novel molecular target to modulate BCRP-mediated MDR.Current evidence suggests that BCRP gene transcription is regulated by a number of trans-acting elements including hypoxia inducible factor 1α,estrogen receptor,and peroxisome proliferator-activated receptor.Furthermore,alternative promoter usage,demethylation of the BCRP promoter,and histone modification are likely associated with drug-induced BCRP overexpression in cancer cells.Finally,PI3K/AKT signaling may play a critical role in modulating BCRP function under a variety of conditions.These biological events seem involved in a complicated manner.Untangling the events would be an essential first step to developing a method to modulate BCRP function to aid patients with cancer.This review will present a synopsis of the impact of BCRP-mediated MDR in cancer cells,and the molecular mechanisms of acquired MDR currently postulated in a variety of human cancers.展开更多
AIM To detect the congenital expression patterns of mdr 1 gene in commonly encountered malignant tumors in clinic, and the relationship between the expression of mdr 1 gene and the prognostic morphology in ...AIM To detect the congenital expression patterns of mdr 1 gene in commonly encountered malignant tumors in clinic, and the relationship between the expression of mdr 1 gene and the prognostic morphology in esophageal carcinomas. METHODS A total of 151 resected samples of malignant tumors without preoperative treatment were taken from Anyang City Tumor Hospital. The congenital expression of their mdr 1 gene was detected with reverse transcription polymerase chain reaction (RT PCR) and was compared with each other. The positive incidence of mdr 1 gene in 46 samples of esophageal carcinoma was compared with their differentiated grades, TNM stages and macroscopic types, and the precautions and advantages of RT PCR were evaluated. RESULTS All the 151 samples were confirmed to be malignant histopathologically, including cancers of stomach and gastric cardia (n =51), esophagus ( n =46), colorectum ( n =16), breast ( n =15), thyroid ( n =10), lung ( n =9), uterine cervix ( n =24). The positive expression rate of their mdr 1 gene was 33 3%, 37%, 31 3%, 13 2%, 40%, 55%, and 0% respectively. All the 46 samples of esophageal carcinoma were pathologically confirmed to be squamous cell carcinoma. The total expression rate of their mdr 1 gene was 37% (17/46), 35% (6/17), 40% (8/20), and 33% (3/9) for differentiation grade Ⅰ, Ⅱ and Ⅲ respectively. The expression rate of TNM classification was 33% (6/18), 40% (5/12) and 37% (6/16) in stage Ⅱa, Ⅱb and Ⅲ. The expression rate was 33% (3/9) in ulcerous type, 37% (3/8) in constrictive one, 33% (5/15) in fungoid one, and 40% (6/14) in medullary one. No statistically significant difference was found. CONCLUSION Compared with other methods, RT PCR is more simple, reliable and accurate in detecting mdr 1 gene expression in tissues of tumor. The overexpression of mdr 1 gene in these neoplasms suggested that cases should be handled differently for chemotherapy with rational use of drugs. Excision is the chief treatment for carcinoma of esophagus. The expression of mdr 1 gene in tissues of esophageal cancer is correlated with the parameters of tumor molecular biology which are independent of histopathological morphology.展开更多
To improve catalytic activity of ribozyme on its substrate, the multi-ribozyme expression system was designed and constructed from 20 cis-acting hammerhead ribozymes undergoing self-cleavage with 10 trans-acting hamme...To improve catalytic activity of ribozyme on its substrate, the multi-ribozyme expression system was designed and constructed from 20 cis-acting hammerhead ribozymes undergoing self-cleavage with 10 trans-acting hammerhead ribozymes inserted alternatively regularly and the plasmid of pGEM-MDR1/MRP1 used to transcribe the MDR1/MRPl（196/210） substrate containing double target sites was also constructed by DNA recombination. Endonuclease digestion analysis and DNA sequencing indicate all the recombinant plasmids were correct. The clea- vage activities were evaluated for the multi-ribozyme expression system on the MDR1/MRP1 substrate in the cell free system. The results demonstrate that the cis-acting hammerhead ribozymes in the multi-ribozyme expression system were able to cleave themselves and the 72 nt of 196Rz and the 71 nt of 210Rz trans-acting hammerhead ribozymes were liberated effectively, and the trans-acting hammerhead ribozymes released were able to act on the MDR1/MRP1 double target RNA substrate and cleave the target RNA at specific sites effectively. The multi- ribozyme expression system of the [Coat＇A196Rz/Coat＇B210Rz]5 is more significantly superior to that of the [Coat＇A 196Rz/Coat＇B210Rz] 1 in cleavage of RNA substrate. The fractions cleaved by [Coat＇A 196Rz/Coat＇B210Rz]5 on the MDR1/MRP1 substrate for 8 h at observed temperatures showed no marked difference. The studies of Mg^2＋ on cleavage efficiency indicate that cleavage reaction is dependent on Mg^2＋ ions concentration. The plot of lg（kobs） vs. lgc（Mg^2＋） displays a linear relationship between 2.5 mmol/L and 20 mmol/L Mg^2＋. It suggests that Mg^2＋ ions play a crucial role in multi-ribozyme cleavage on the substrate.展开更多
MALIGNANT tumor is very harmful to human health, and the mortality is very high. About 50% of the patients with malignancy can be operated on, and the other 50% patients have to be treated with chemotherapy. Because t...MALIGNANT tumor is very harmful to human health, and the mortality is very high. About 50% of the patients with malignancy can be operated on, and the other 50% patients have to be treated with chemotherapy. Because tumors are mostly chemoresistant, chemotherapy for these patients often has no effect. The overexpression of MDR1 gene is very common in hu man malignant tumors, about 50% in previously untreated patients and more than 50% in previously treated patients for whom the tumor is resistant to the previous sensitive展开更多
AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation mark...AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation marker of BCRP expression.METHODS MTT assay was used to filtrate BCRP-mediatedresistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated withdifferent concentration anticancer agents.High performance liquid chromatography(HPLC) was applied tomeasure relative dose of intracellular retention resistance agents.Nuclear DNA fluorescence dye,Hochest33258,staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs.RESULTSThere were shown increasing durg-resistance to 5-fluorouracil,methotrexate,doxirubicin,pirarubicin,etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRPcells(P<0.05,n=3),but shown sensitive to paclitaxel,cisplatin,vincristine,mitomycin and vindesine.Therealso was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil withdifferent expression of BCRP(r=-0.885,P<0.05,n=3).There were shown parallel results ofthat decreasingcellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil byfluorescence dye staining and flow cytometry(P<0.05,n=3),and also shown significate rise of the apoptoticrate of BCRP expression cells after treated with Ko143 (P<0.05,n=3).Every group of cells could be differentextently blocked in phase of G_0/G_1 treated with 5-fluorouracil.CONCLUSION Resistance of 5-fluorouracilcould be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate.Cellularability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.展开更多
文摘Breast cancer resistance protein(BCRP)/ATP-binding cassette subfamily G member 2(ABCG2) is an ATP-binding cassette(ABC) transporter identified as a molecular cause of multidrug resistance(MDR) in diverse cancer cells.BCRP physiologically functions as a part of a self-defense mechanism for the organism;it enhances elimination of toxic xenobiotic substances and harmful agents in the gut and biliary tract,as well as through the blood-brain,placental,and possibly blood-testis barriers.BCRP recognizes and transports numerous anticancer drugs including conventional chemotherapeutic and targeted small therapeutic molecules relatively new in clinical use.Thus,BCRP expression in cancer cells directly causes MDR by active efflux of anticancer drugs.Because BCRP is also known to be a stem cell marker,its expression in cancer cells could be a manifestation of metabolic and signaling pathways that confer multiple mechanisms of drug resistance,self-renewal(stemness),and invasiveness(aggressiveness),and thereby impart a poor prognosis.Therefore,blocking BCRP-mediated active efflux may provide a therapeutic benefit for cancers.Delineating the precise molecular mechanisms for BCRP gene expression may lead to identification of a novel molecular target to modulate BCRP-mediated MDR.Current evidence suggests that BCRP gene transcription is regulated by a number of trans-acting elements including hypoxia inducible factor 1α,estrogen receptor,and peroxisome proliferator-activated receptor.Furthermore,alternative promoter usage,demethylation of the BCRP promoter,and histone modification are likely associated with drug-induced BCRP overexpression in cancer cells.Finally,PI3K/AKT signaling may play a critical role in modulating BCRP function under a variety of conditions.These biological events seem involved in a complicated manner.Untangling the events would be an essential first step to developing a method to modulate BCRP function to aid patients with cancer.This review will present a synopsis of the impact of BCRP-mediated MDR in cancer cells,and the molecular mechanisms of acquired MDR currently postulated in a variety of human cancers.
文摘AIM To detect the congenital expression patterns of mdr 1 gene in commonly encountered malignant tumors in clinic, and the relationship between the expression of mdr 1 gene and the prognostic morphology in esophageal carcinomas. METHODS A total of 151 resected samples of malignant tumors without preoperative treatment were taken from Anyang City Tumor Hospital. The congenital expression of their mdr 1 gene was detected with reverse transcription polymerase chain reaction (RT PCR) and was compared with each other. The positive incidence of mdr 1 gene in 46 samples of esophageal carcinoma was compared with their differentiated grades, TNM stages and macroscopic types, and the precautions and advantages of RT PCR were evaluated. RESULTS All the 151 samples were confirmed to be malignant histopathologically, including cancers of stomach and gastric cardia (n =51), esophagus ( n =46), colorectum ( n =16), breast ( n =15), thyroid ( n =10), lung ( n =9), uterine cervix ( n =24). The positive expression rate of their mdr 1 gene was 33 3%, 37%, 31 3%, 13 2%, 40%, 55%, and 0% respectively. All the 46 samples of esophageal carcinoma were pathologically confirmed to be squamous cell carcinoma. The total expression rate of their mdr 1 gene was 37% (17/46), 35% (6/17), 40% (8/20), and 33% (3/9) for differentiation grade Ⅰ, Ⅱ and Ⅲ respectively. The expression rate of TNM classification was 33% (6/18), 40% (5/12) and 37% (6/16) in stage Ⅱa, Ⅱb and Ⅲ. The expression rate was 33% (3/9) in ulcerous type, 37% (3/8) in constrictive one, 33% (5/15) in fungoid one, and 40% (6/14) in medullary one. No statistically significant difference was found. CONCLUSION Compared with other methods, RT PCR is more simple, reliable and accurate in detecting mdr 1 gene expression in tissues of tumor. The overexpression of mdr 1 gene in these neoplasms suggested that cases should be handled differently for chemotherapy with rational use of drugs. Excision is the chief treatment for carcinoma of esophagus. The expression of mdr 1 gene in tissues of esophageal cancer is correlated with the parameters of tumor molecular biology which are independent of histopathological morphology.
基金Supported by Fund of Shenzhen Bureau of Science and Technology, China(No.20008).
文摘To improve catalytic activity of ribozyme on its substrate, the multi-ribozyme expression system was designed and constructed from 20 cis-acting hammerhead ribozymes undergoing self-cleavage with 10 trans-acting hammerhead ribozymes inserted alternatively regularly and the plasmid of pGEM-MDR1/MRP1 used to transcribe the MDR1/MRPl（196/210） substrate containing double target sites was also constructed by DNA recombination. Endonuclease digestion analysis and DNA sequencing indicate all the recombinant plasmids were correct. The clea- vage activities were evaluated for the multi-ribozyme expression system on the MDR1/MRP1 substrate in the cell free system. The results demonstrate that the cis-acting hammerhead ribozymes in the multi-ribozyme expression system were able to cleave themselves and the 72 nt of 196Rz and the 71 nt of 210Rz trans-acting hammerhead ribozymes were liberated effectively, and the trans-acting hammerhead ribozymes released were able to act on the MDR1/MRP1 double target RNA substrate and cleave the target RNA at specific sites effectively. The multi- ribozyme expression system of the [Coat＇A196Rz/Coat＇B210Rz]5 is more significantly superior to that of the [Coat＇A 196Rz/Coat＇B210Rz] 1 in cleavage of RNA substrate. The fractions cleaved by [Coat＇A 196Rz/Coat＇B210Rz]5 on the MDR1/MRP1 substrate for 8 h at observed temperatures showed no marked difference. The studies of Mg^2＋ on cleavage efficiency indicate that cleavage reaction is dependent on Mg^2＋ ions concentration. The plot of lg（kobs） vs. lgc（Mg^2＋） displays a linear relationship between 2.5 mmol/L and 20 mmol/L Mg^2＋. It suggests that Mg^2＋ ions play a crucial role in multi-ribozyme cleavage on the substrate.
文摘MALIGNANT tumor is very harmful to human health, and the mortality is very high. About 50% of the patients with malignancy can be operated on, and the other 50% patients have to be treated with chemotherapy. Because tumors are mostly chemoresistant, chemotherapy for these patients often has no effect. The overexpression of MDR1 gene is very common in hu man malignant tumors, about 50% in previously untreated patients and more than 50% in previously treated patients for whom the tumor is resistant to the previous sensitive
文摘AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation marker of BCRP expression.METHODS MTT assay was used to filtrate BCRP-mediatedresistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated withdifferent concentration anticancer agents.High performance liquid chromatography(HPLC) was applied tomeasure relative dose of intracellular retention resistance agents.Nuclear DNA fluorescence dye,Hochest33258,staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs.RESULTSThere were shown increasing durg-resistance to 5-fluorouracil,methotrexate,doxirubicin,pirarubicin,etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRPcells(P<0.05,n=3),but shown sensitive to paclitaxel,cisplatin,vincristine,mitomycin and vindesine.Therealso was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil withdifferent expression of BCRP(r=-0.885,P<0.05,n=3).There were shown parallel results ofthat decreasingcellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil byfluorescence dye staining and flow cytometry(P<0.05,n=3),and also shown significate rise of the apoptoticrate of BCRP expression cells after treated with Ko143 (P<0.05,n=3).Every group of cells could be differentextently blocked in phase of G_0/G_1 treated with 5-fluorouracil.CONCLUSION Resistance of 5-fluorouracilcould be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate.Cellularability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.
