The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation,differentiation,invasion and metastasis of cancer cells,but its role in cervical cancer is still u...The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation,differentiation,invasion and metastasis of cancer cells,but its role in cervical cancer is still unclear.This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors.The NDRG1 expression in cervical tissues and cells was detected by RT-PCR.Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines.The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting,respectively.Its effects on cell proliferation,migration,invasion,cell cycle and apoptosis were detected by MTT,transwell migration assay and flow cytometry (FCM),respectively.The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001).After transfection with pEGFP-N1-NDRG1-GFP,the mRNA and protein expression of NDRG1 was up-regulated in Siha cells,which suppressed cell proliferation (P<0.001),induced cell cycle arrest (P<0.05),reduced invasion and migration of Siha cells (P<0.05),but caused no cell apoptosis.Moreover,vascular endothelial growth factor (VEGF),a tumor-induced angiogenesis factor,was markedly reduced and E-cadherin,a cell adhesion molecule,was increased in the cells transfected with pEGFP-N1-NDRG1-GFP.It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth,invasion and metastasis of human cervical cancer.展开更多
Prolactinoma is an estrogen‐related tumor and leukemia‐related protein 16(LRP16)is correlated with the progression of estrogen‐related tumors,but the regulatory mechanism between LRP16 and prolactinoma remain uncle...Prolactinoma is an estrogen‐related tumor and leukemia‐related protein 16(LRP16)is correlated with the progression of estrogen‐related tumors,but the regulatory mechanism between LRP16 and prolactinoma remain unclear.This study demonstrates a variation in LRP16 with estrogen receptorα(ERα)in prolactinoma models and the up and downregulation effects of LRP16 on prolactin secretion of pituitary adenomas cells(GH_3cells).In our study,50 male SD rats(30‐day‐old)展开更多
:AIM To construct Hsp90 antisense RNAeukaryotic expression vector, transfect it intoSGC7901 and SGC7901/VCR of MDR-type humangastric cancer cell lines, HCC7402 of humanhepatic cancer and Eel09 of human esophagealcance...:AIM To construct Hsp90 antisense RNAeukaryotic expression vector, transfect it intoSGC7901 and SGC7901/VCR of MDR-type humangastric cancer cell lines, HCC7402 of humanhepatic cancer and Eel09 of human esophagealcancer cell lines, and to study the cell cycledistribution of the gene transected cells andtheir response to chemotherapeutic drugs.METHODS A I .03kb cDNA sequence of Hsp90Pwas obtained from the primary plasmid phHsp90by EcoR 1 and BamH I nuclease digestion andwas cloned to the EcoR 1 and BamH 1 site ofthe pcDNA by T4DNA ligase and an antisenseorientation of Hsp900 expression vector wasconstructed. The constructs were transfectedwith lipofectamine and positive clones wereselected with G418. The expression of RNA wasdetermined with dot blotting and RNaseprotection assay, and the expression of Hsp90protein determined with Western blot. Cell cycledistribution of the transfectants was analyzedwith flow cytometry, and the drug sensitivity ofthe transfectants to adriamycin (ADR ),vincrinstine (VCR ), mitomycin (MMC ) andcyclophosphamide (CTX ) with MTT andintracellular drug concentration of thetransfectants was determined with flowcytometry.RESULTS In EcoR 1 and BamH I restrictionanalysis, the size and the direction of the clonedsequence of Hsp900 remained what had beendesigned and the gene constructs were namedpcDNA-Hsp90. AH^SGC7901, AH^SGC7901/ VCR,AH-HCC7402 and AH-Eel09 cell clones allexpressed Hsp90 anti--sense RNA. Theexpression of Hsp90 was down--regulated in AHSGC7901, AH--SGC7901/ VCR, AH-HCC7402 andAH--Eel09 cell clones. Cell cycle distribution waschanged differently. In AH-SGC7901/ VCR andAH-Ec109 cells, G, phase cells were increased; Sphase and G, phase cells were decreased ascompared with their parental cell lines. In AHSGC7901 cell, G, phase cells were decreased, Qphase cells increased and S phase cells were notchanged, and in AH-HCC7402 cells G,, S and qphase cells remained unchanged as comparedwith their parental cell lines. The sensitivity ofAH--SGC7901, AH--SGC7901/ VCR, AH-HCC7402 andAH-Ec109 to chemotherapeutic drugs, thesensitivity ot AH--SGC7901/ VCR to ADR, VCR,MMC and CTX the sensitivity of AH-HCC7402 toADR and VCR, and the sensitivity of Eel09 toADR, VCR and CTX all increased as comparedwith their parental cell lines. The meanfluorescence intensity of ADR in AH--SGC7901,AH-SGC7901/ VCR, AH--HCC7402 and AH-Ec109was also significantly elevated (P< 0. 05).CONCLUSION Down-regulation of HsP90 couldchange cell cycle distribution and increase thedrug sensitivity of tumor cells.展开更多
Particulate matter （PM）, which is a greatenvironmental concern, has been classified as aGroup 1 human carcinogen by the InternationalAgency for Research on Cancer （IARC）.Epidemiological and experimental studies ha...Particulate matter （PM）, which is a greatenvironmental concern, has been classified as aGroup 1 human carcinogen by the InternationalAgency for Research on Cancer （IARC）.Epidemiological and experimental studies haveindicated that chronic exposure to PM, especiallyPM2.5 （particles with an aerodynamic diameter lessthan 2.5 μm） may lead to an increase in lung cancerincidence.展开更多
The effect of glucocorticoids on the down-regulation of glucocorticoidreceptor (GR) mRNA was studied in intact rats.GR mRNA was characterized byNorthern blot hybridization and quantitated by dot blot hybridization usi...The effect of glucocorticoids on the down-regulation of glucocorticoidreceptor (GR) mRNA was studied in intact rats.GR mRNA was characterized byNorthern blot hybridization and quantitated by dot blot hybridization using a hu-man GR cDNA fragment as a probe.Administration of hydrocortisone (F) inpolyvinyl alcohol (PVA) resulted in a rapid increase in plasma glucocorticoidswhich maintained at stress levels (20 to 40μg/dl) for about 3 d.HepaticGR mRNA decreased significantly to 73.5±6.3% of control values 6h followingF treatment,after which the decline of GR mRNA was gradual,reaching a mini-mum of 44.0±5.0% of control levels 3d after the treatment.The effect of F onthe down-regulation of hepatic GR mRNA lasted up to 11 d.In contrast,F treat-ment had no effect on GR mRNA in rat brain.These results are consistent withthe changes in GR in rats as reported previously,except that even though thehepatic cytosol GR decreased markedly,no significant changes in hepatic GRmRNA were found 1h after F treatment,strongly suggesting that thedown-regulation of GR by its ligands in vivo occurs at both transcriptional andposttranscriptional levels and is of tissue-specific fashion.展开更多
Lead exposure is a known potential risk factor for neurodegenerative diseases such as Alzheimer's disease(AD). Exposure to lead during the critical phase of brain development has been linked with mental retardatio...Lead exposure is a known potential risk factor for neurodegenerative diseases such as Alzheimer's disease(AD). Exposure to lead during the critical phase of brain development has been linked with mental retardation and hypophrenia in later life. This study was aimed to investigate the effects of lead exposure of pregnant mice on the expressions of insulin-degrading enzyme(IDE) and nerve growth factor(NGF) in the hippocampus of their offspring. Blood samples were collected from the tail vein, and after anesthetizing the pups, the brain was excised on postnatal day 21. Lead concentrations were determined by graphite furnace atomic absorption spectrophotometry, and the expressions of IDE and NGF were determined by immunohistochemistry and Western blotting. Results showed that the reduction in IDE and NGF expression in the hippocampus of pups might be associated with impairment of learning and memory and dementia induced by maternal lead exposure during pregnancy and lactation.展开更多
Objective To study the inhibitory effect of siRNA on glioblastoma (GBM) Notch-1 gene expression in addition to the growth of TJ-905 glioblastoma. Methods Three small interference RNAs (siRNAs) targeting Notch1 gene,na...Objective To study the inhibitory effect of siRNA on glioblastoma (GBM) Notch-1 gene expression in addition to the growth of TJ-905 glioblastoma. Methods Three small interference RNAs (siRNAs) targeting Notch1 gene,named siRNA1,siRNA2,siRNA3,synthesized chemically in vitro with gene bank BLAST. TJ-905 cells were transfected twice with the siRNA by using Oligofectamine展开更多
The regulatory mechanisms of drought and salt-associated miRNAs have not been fully understood in Sorghum bicolor. In this study, we investigated the effect of salinity stress (200 and 300 mM NaCl) and drought stress ...The regulatory mechanisms of drought and salt-associated miRNAs have not been fully understood in Sorghum bicolor. In this study, we investigated the effect of salinity stress (200 and 300 mM NaCl) and drought stress at pre- and post-flowering stages on the expression pattern of small regulatory RNAs in six Sorghum genotypes using semi-quantitative reverse transcriptase PCR (RT-qPCR). The results indicated that both drought and salt stresses altered the expression pattern of miRNAs in a dose-dependent manner. However, each miRNA responded to drought and salt stress in a different pattern among the six sorghum genotypes. miR156, miR167, miR168 and miR399 give different expressions levels compared to other studied miRNAs which may attribute to the adaption of sorghum to drought and salt stress and are good candidates for improving sorghum by transgenic technology.展开更多
Adenomatosis polyposis down-regulated 1(APCDD1) is a transmembrane glycoprotein that negatively regulates Wnt/β-catenin canonical signaling by binding with Wnt ligands and receptors. We analyzed the role of APCDD1 in...Adenomatosis polyposis down-regulated 1(APCDD1) is a transmembrane glycoprotein that negatively regulates Wnt/β-catenin canonical signaling by binding with Wnt ligands and receptors. We analyzed the role of APCDD1 in the Wnt5a/c-Jun non-canonical signaling pathway and demonstrated that APCDD1 can interact in vitro with Wnt5a, a classical ligand, and Ror2, a receptor of non-canonical Wnt signaling. Furthermore, we verified the binding of APCDD1 and Ror2 in primary cells of mouse skin. Moreover, APCDD1 seems to form a complex with Ror2 and Vangl2 in the cell, and complex formation can be improved by adding Wnt5a. In the presence of Wnt5a and Ror2, APCDD1 can induce the phosphorylation of c-Jun, a transcription factor of Wnt5a non-canonical signaling, and its phosphorylation level is a readout of Wnt5 a signaling. Wound-healing assay shows that APCDD1 accelerates polarized cell migration during Wnt5a-induced wound closure. Therefore,it is very likely that APCDD1 regulates Wnt5a/c-Jun non-canonical signaling as co-receptor binding with both Wnt5a and Ror2.展开更多
基金supported by a grant from the Natural Sciences Foundation of Hubei Province(No.4-306)
文摘The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation,differentiation,invasion and metastasis of cancer cells,but its role in cervical cancer is still unclear.This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors.The NDRG1 expression in cervical tissues and cells was detected by RT-PCR.Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines.The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting,respectively.Its effects on cell proliferation,migration,invasion,cell cycle and apoptosis were detected by MTT,transwell migration assay and flow cytometry (FCM),respectively.The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001).After transfection with pEGFP-N1-NDRG1-GFP,the mRNA and protein expression of NDRG1 was up-regulated in Siha cells,which suppressed cell proliferation (P<0.001),induced cell cycle arrest (P<0.05),reduced invasion and migration of Siha cells (P<0.05),but caused no cell apoptosis.Moreover,vascular endothelial growth factor (VEGF),a tumor-induced angiogenesis factor,was markedly reduced and E-cadherin,a cell adhesion molecule,was increased in the cells transfected with pEGFP-N1-NDRG1-GFP.It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth,invasion and metastasis of human cervical cancer.
基金financially supported by the National Natural Science Foundation of China and Hainan Province.[NSFC,Grant Nos.81570705 and 81270866NSFH,Grant No.20168353]
文摘Prolactinoma is an estrogen‐related tumor and leukemia‐related protein 16(LRP16)is correlated with the progression of estrogen‐related tumors,but the regulatory mechanism between LRP16 and prolactinoma remain unclear.This study demonstrates a variation in LRP16 with estrogen receptorα(ERα)in prolactinoma models and the up and downregulation effects of LRP16 on prolactin secretion of pituitary adenomas cells(GH_3cells).In our study,50 male SD rats(30‐day‐old)
基金Project supported by the National Natural Science Foundation of China,No.39570806National Excel1ent Youth Scientific Foundation,No.3952020.
文摘:AIM To construct Hsp90 antisense RNAeukaryotic expression vector, transfect it intoSGC7901 and SGC7901/VCR of MDR-type humangastric cancer cell lines, HCC7402 of humanhepatic cancer and Eel09 of human esophagealcancer cell lines, and to study the cell cycledistribution of the gene transected cells andtheir response to chemotherapeutic drugs.METHODS A I .03kb cDNA sequence of Hsp90Pwas obtained from the primary plasmid phHsp90by EcoR 1 and BamH I nuclease digestion andwas cloned to the EcoR 1 and BamH 1 site ofthe pcDNA by T4DNA ligase and an antisenseorientation of Hsp900 expression vector wasconstructed. The constructs were transfectedwith lipofectamine and positive clones wereselected with G418. The expression of RNA wasdetermined with dot blotting and RNaseprotection assay, and the expression of Hsp90protein determined with Western blot. Cell cycledistribution of the transfectants was analyzedwith flow cytometry, and the drug sensitivity ofthe transfectants to adriamycin (ADR ),vincrinstine (VCR ), mitomycin (MMC ) andcyclophosphamide (CTX ) with MTT andintracellular drug concentration of thetransfectants was determined with flowcytometry.RESULTS In EcoR 1 and BamH I restrictionanalysis, the size and the direction of the clonedsequence of Hsp900 remained what had beendesigned and the gene constructs were namedpcDNA-Hsp90. AH^SGC7901, AH^SGC7901/ VCR,AH-HCC7402 and AH-Eel09 cell clones allexpressed Hsp90 anti--sense RNA. Theexpression of Hsp90 was down--regulated in AHSGC7901, AH--SGC7901/ VCR, AH-HCC7402 andAH--Eel09 cell clones. Cell cycle distribution waschanged differently. In AH-SGC7901/ VCR andAH-Ec109 cells, G, phase cells were increased; Sphase and G, phase cells were decreased ascompared with their parental cell lines. In AHSGC7901 cell, G, phase cells were decreased, Qphase cells increased and S phase cells were notchanged, and in AH-HCC7402 cells G,, S and qphase cells remained unchanged as comparedwith their parental cell lines. The sensitivity ofAH--SGC7901, AH--SGC7901/ VCR, AH-HCC7402 andAH-Ec109 to chemotherapeutic drugs, thesensitivity ot AH--SGC7901/ VCR to ADR, VCR,MMC and CTX the sensitivity of AH-HCC7402 toADR and VCR, and the sensitivity of Eel09 toADR, VCR and CTX all increased as comparedwith their parental cell lines. The meanfluorescence intensity of ADR in AH--SGC7901,AH-SGC7901/ VCR, AH--HCC7402 and AH-Ec109was also significantly elevated (P< 0. 05).CONCLUSION Down-regulation of HsP90 couldchange cell cycle distribution and increase thedrug sensitivity of tumor cells.
基金supported by the National Natural Science Foundation of China[81202231 to LLH]the Medical Scientific Research Funding of Guangdong Province,China[A2018225 to LLH]+1 种基金the College Students Cultivate Special Science and Technology Innovation from Education Department of Guangdong Province,China[pdjh2016a0212]the Project for Creative Talent of Guangdong Education Department[2014KQNCX102]
文摘Particulate matter （PM）, which is a greatenvironmental concern, has been classified as aGroup 1 human carcinogen by the InternationalAgency for Research on Cancer （IARC）.Epidemiological and experimental studies haveindicated that chronic exposure to PM, especiallyPM2.5 （particles with an aerodynamic diameter lessthan 2.5 μm） may lead to an increase in lung cancerincidence.
文摘The effect of glucocorticoids on the down-regulation of glucocorticoidreceptor (GR) mRNA was studied in intact rats.GR mRNA was characterized byNorthern blot hybridization and quantitated by dot blot hybridization using a hu-man GR cDNA fragment as a probe.Administration of hydrocortisone (F) inpolyvinyl alcohol (PVA) resulted in a rapid increase in plasma glucocorticoidswhich maintained at stress levels (20 to 40μg/dl) for about 3 d.HepaticGR mRNA decreased significantly to 73.5±6.3% of control values 6h followingF treatment,after which the decline of GR mRNA was gradual,reaching a mini-mum of 44.0±5.0% of control levels 3d after the treatment.The effect of F onthe down-regulation of hepatic GR mRNA lasted up to 11 d.In contrast,F treat-ment had no effect on GR mRNA in rat brain.These results are consistent withthe changes in GR in rats as reported previously,except that even though thehepatic cytosol GR decreased markedly,no significant changes in hepatic GRmRNA were found 1h after F treatment,strongly suggesting that thedown-regulation of GR by its ligands in vivo occurs at both transcriptional andposttranscriptional levels and is of tissue-specific fashion.
基金supported by the National Natural Science Foundation of China(NSFC),No.31201878,81172716,and U1204804Post Doctoral Foundation of China,No.2015M572109Post Doctoral Fund of Henan province,No.2014049
文摘Lead exposure is a known potential risk factor for neurodegenerative diseases such as Alzheimer's disease(AD). Exposure to lead during the critical phase of brain development has been linked with mental retardation and hypophrenia in later life. This study was aimed to investigate the effects of lead exposure of pregnant mice on the expressions of insulin-degrading enzyme(IDE) and nerve growth factor(NGF) in the hippocampus of their offspring. Blood samples were collected from the tail vein, and after anesthetizing the pups, the brain was excised on postnatal day 21. Lead concentrations were determined by graphite furnace atomic absorption spectrophotometry, and the expressions of IDE and NGF were determined by immunohistochemistry and Western blotting. Results showed that the reduction in IDE and NGF expression in the hippocampus of pups might be associated with impairment of learning and memory and dementia induced by maternal lead exposure during pregnancy and lactation.
文摘Objective To study the inhibitory effect of siRNA on glioblastoma (GBM) Notch-1 gene expression in addition to the growth of TJ-905 glioblastoma. Methods Three small interference RNAs (siRNAs) targeting Notch1 gene,named siRNA1,siRNA2,siRNA3,synthesized chemically in vitro with gene bank BLAST. TJ-905 cells were transfected twice with the siRNA by using Oligofectamine
文摘The regulatory mechanisms of drought and salt-associated miRNAs have not been fully understood in Sorghum bicolor. In this study, we investigated the effect of salinity stress (200 and 300 mM NaCl) and drought stress at pre- and post-flowering stages on the expression pattern of small regulatory RNAs in six Sorghum genotypes using semi-quantitative reverse transcriptase PCR (RT-qPCR). The results indicated that both drought and salt stresses altered the expression pattern of miRNAs in a dose-dependent manner. However, each miRNA responded to drought and salt stress in a different pattern among the six sorghum genotypes. miR156, miR167, miR168 and miR399 give different expressions levels compared to other studied miRNAs which may attribute to the adaption of sorghum to drought and salt stress and are good candidates for improving sorghum by transgenic technology.
基金the National Natural Science Foundation of China(Nos.31671504 and 81421061)the Cross Research Fund of Biomedical Engineering of Shanghai Jiao Tong University(No.YG2016MS04)+1 种基金the Natural Science Foundation of Shanghai(No.13ZR1421100)the National Key Technology R&D Program of China(No.2012BAI01B09)
文摘Adenomatosis polyposis down-regulated 1(APCDD1) is a transmembrane glycoprotein that negatively regulates Wnt/β-catenin canonical signaling by binding with Wnt ligands and receptors. We analyzed the role of APCDD1 in the Wnt5a/c-Jun non-canonical signaling pathway and demonstrated that APCDD1 can interact in vitro with Wnt5a, a classical ligand, and Ror2, a receptor of non-canonical Wnt signaling. Furthermore, we verified the binding of APCDD1 and Ror2 in primary cells of mouse skin. Moreover, APCDD1 seems to form a complex with Ror2 and Vangl2 in the cell, and complex formation can be improved by adding Wnt5a. In the presence of Wnt5a and Ror2, APCDD1 can induce the phosphorylation of c-Jun, a transcription factor of Wnt5a non-canonical signaling, and its phosphorylation level is a readout of Wnt5 a signaling. Wound-healing assay shows that APCDD1 accelerates polarized cell migration during Wnt5a-induced wound closure. Therefore,it is very likely that APCDD1 regulates Wnt5a/c-Jun non-canonical signaling as co-receptor binding with both Wnt5a and Ror2.
