Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted ...Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.展开更多
Background The 5-hydroxytryptamine7 receptor （5-HT7 receptor, 5-HTTR） plays an important role in the regulation of smooth muscle relaxation and visceral sensation and might be involved in the pathogenesis of the gas...Background The 5-hydroxytryptamine7 receptor （5-HT7 receptor, 5-HTTR） plays an important role in the regulation of smooth muscle relaxation and visceral sensation and might be involved in the pathogenesis of the gastrointestinal dyskinesia, abdominal pain and visceral paresthesia in irritable bowel syndrome （IBS）. The aim of this study was to investigate the role of the 5-HT7 receptor in the pathogenesis of IBS. Methods A rat model of irritable bowel syndrome with diarrhea （IBS-D） was established by colonic instillation of acetic acid and restraint stress. A rat model with irritable bowel syndrome with constipation （IBS-C） was established by stomach irrigated with 0-4℃ cool water daily for 14 days. The content and distribution of 5-HT in the brain and gut were examined by immunohistochemistry and the mRNA expression of the 5-HT7 receptor was determined by fluorescent quantitative reverse transcription polymerase chain reaction. The accumulation of cyclic adenosine monophosphate （cAMP） in all the same tissues was measured by radioimmunity. Results The models of IBS were reliable by identification. The immunohistochemistry results showed that there were significantly more 5-HT positive cells in the IBS-D group than in the control group in the hippocampus, hypothalamus, jejunum, ileum, proximate colon and distal colon （P〈0.05）, as well as more than were found in the IBS-C group in jejunum and ileum （P〈0.05）. There were more 5-HT positive cells in the IBS-C group than in the control hippocampus, hypothalamus, ileum, proximate colon, and distal colon （P〈0.05）. Real time-PCR results showed that the expression level of the 5-HT7 receptor in both the IBS-C and IBS-D groups were enhanced compared with the control group in the hippocampus and hypothalamus （P〈0.05）. The expression level of 5-HT7 receptors in the IBS-C group was notably greater when compared with the controls in the ileum and colon （P〈0.05）. The cAMP accumulation in the hippocampus and hypothalamus in both the IBS-C and IBS-D groups was higher than that in the control group （P〈0.01 and P〈0.05）. The cAMP accumulation in the tBS-C group was higher than that in the control group in the proximal and distal colon （m〈o.o5）. Conclusions The increased 5-HT content in the brain and intestine is related to the IBS pathogenesis. The up-regulated expression of the 5-HT7 receptor in the brain and colon might play an important role in the pathogenesis of IBS-C.展开更多
Objective：To investigate the role of 5-HT7 receptor in the pathogenesis of irritable bowel syndrome（IBS）. Methods：Rat model of D-IBS was established by intracolonic instillation of acetic acid and restraint stress...Objective：To investigate the role of 5-HT7 receptor in the pathogenesis of irritable bowel syndrome（IBS）. Methods：Rat model of D-IBS was established by intracolonic instillation of acetic acid and restraint stress; Rat model of C-IBS was established by stomach irrigated with 0-4℃ cool water daily for 14 d. The content and distribution of 5-HT7 receptor at the brain and bowel was examined by immunohistochemistry and the expression of 5-HT7 receptor mRNA was detected by fluorescence quantitative RT-PCR（Real-time PCR）. Results：Immunocytochemistry result showed the 5-HT7 rceptor positive staining at hippocampus and hypothalamus of both C-IBS and D-IBS group was stronger than that of control group（P 〈 0.01）. The 5-HT7R expression at ileum, proximate colon, distal colon of C-IBS group was significantly stronger than that of control group（P 〈 0.05）. Realtime-PCR analysis results showed the expression level of 5-HT7 receptor at hippocampus and hypothalamus of both C-IBS and D-IBS group was increased than that of control group（P〈 0.05）. At proximal and distal colon of C-IBS group, the 5-HT7 receptor mRNA expression was increased compared with control group（P〈 0.05）. Conclusion：The up-regulated expression of 5-HT7 receptor at brain and colon may play an important role in the pathogenesis of C-IBS.展开更多
It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and ...It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.展开更多
It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The a...It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2c receptor gene polymorphism and the duration of IELT in men with LPE. Therefore, a prospective study was conducted in 64 Dutch Caucasian men with LPE. Baseline IELT during coitus was assessed by stopwatch over a 1-month period. All men were genotyped for Cys23Ser 5-HT2c receptor gene polymorphism. Allele frequencies and genotypes of Cys and Ser variants of 5-HT2c receptor gene polymorphism were determined. Association between Cys/Cys and Ser/Ser genotypes and the natural logarithm of the IELT in men with LPE were.investigated. As a result, the geometric mean, median and natural mean IELT were 25.2, 27.0, 33.9s, respectively. Of all men, 20.0%, 10.8%, 23.1% and 41.5% ejaculated within 10, 10-20, 20-30 and 30-60s after vaginal penetration. Of the 64 men, the Cys/Cys and Ser/Ser genotype frequency for the Cys23Ser polymorphism of the 5-HT2c receptor gene was 81% and 19%, respectively. The geometric mean IELT of the wildtypes （Cys/Cys） is significantly lower （22.6s; 95% CI 18.3-27.8s） than in male homozygous mutants （Ser/Ser） （40.4s; 95% CI 20.3-80.4s） （P = 0.03）. It is concluded that Cys23Ser 5-HT2c receptor gene polymorphism is associated with the IELT in men with LPE. Men with Cys/Cys genotype have shorter IELTs than men with Ser/Ser genotypes.展开更多
文摘Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.
基金the key clinical project of Chinese Ministry of Health(2004-2006).
文摘Background The 5-hydroxytryptamine7 receptor （5-HT7 receptor, 5-HTTR） plays an important role in the regulation of smooth muscle relaxation and visceral sensation and might be involved in the pathogenesis of the gastrointestinal dyskinesia, abdominal pain and visceral paresthesia in irritable bowel syndrome （IBS）. The aim of this study was to investigate the role of the 5-HT7 receptor in the pathogenesis of IBS. Methods A rat model of irritable bowel syndrome with diarrhea （IBS-D） was established by colonic instillation of acetic acid and restraint stress. A rat model with irritable bowel syndrome with constipation （IBS-C） was established by stomach irrigated with 0-4℃ cool water daily for 14 days. The content and distribution of 5-HT in the brain and gut were examined by immunohistochemistry and the mRNA expression of the 5-HT7 receptor was determined by fluorescent quantitative reverse transcription polymerase chain reaction. The accumulation of cyclic adenosine monophosphate （cAMP） in all the same tissues was measured by radioimmunity. Results The models of IBS were reliable by identification. The immunohistochemistry results showed that there were significantly more 5-HT positive cells in the IBS-D group than in the control group in the hippocampus, hypothalamus, jejunum, ileum, proximate colon and distal colon （P〈0.05）, as well as more than were found in the IBS-C group in jejunum and ileum （P〈0.05）. There were more 5-HT positive cells in the IBS-C group than in the control hippocampus, hypothalamus, ileum, proximate colon, and distal colon （P〈0.05）. Real time-PCR results showed that the expression level of the 5-HT7 receptor in both the IBS-C and IBS-D groups were enhanced compared with the control group in the hippocampus and hypothalamus （P〈0.05）. The expression level of 5-HT7 receptors in the IBS-C group was notably greater when compared with the controls in the ileum and colon （P〈0.05）. The cAMP accumulation in the hippocampus and hypothalamus in both the IBS-C and IBS-D groups was higher than that in the control group （P〈0.01 and P〈0.05）. The cAMP accumulation in the tBS-C group was higher than that in the control group in the proximal and distal colon （m〈o.o5）. Conclusions The increased 5-HT content in the brain and intestine is related to the IBS pathogenesis. The up-regulated expression of the 5-HT7 receptor in the brain and colon might play an important role in the pathogenesis of IBS-C.
基金This study was supported by grants from the the National Natural Science Foundation of China(No.30170414)
文摘Objective：To investigate the role of 5-HT7 receptor in the pathogenesis of irritable bowel syndrome（IBS）. Methods：Rat model of D-IBS was established by intracolonic instillation of acetic acid and restraint stress; Rat model of C-IBS was established by stomach irrigated with 0-4℃ cool water daily for 14 d. The content and distribution of 5-HT7 receptor at the brain and bowel was examined by immunohistochemistry and the expression of 5-HT7 receptor mRNA was detected by fluorescence quantitative RT-PCR（Real-time PCR）. Results：Immunocytochemistry result showed the 5-HT7 rceptor positive staining at hippocampus and hypothalamus of both C-IBS and D-IBS group was stronger than that of control group（P 〈 0.01）. The 5-HT7R expression at ileum, proximate colon, distal colon of C-IBS group was significantly stronger than that of control group（P 〈 0.05）. Realtime-PCR analysis results showed the expression level of 5-HT7 receptor at hippocampus and hypothalamus of both C-IBS and D-IBS group was increased than that of control group（P〈 0.05）. At proximal and distal colon of C-IBS group, the 5-HT7 receptor mRNA expression was increased compared with control group（P〈 0.05）. Conclusion：The up-regulated expression of 5-HT7 receptor at brain and colon may play an important role in the pathogenesis of C-IBS.
基金National Natural Science Foundation of China(Grant No.30772556 and 30640070)Research Fund of Janssen Research Council and the‘985'Project in Peking University.
文摘It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.
文摘It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2c receptor gene polymorphism and the duration of IELT in men with LPE. Therefore, a prospective study was conducted in 64 Dutch Caucasian men with LPE. Baseline IELT during coitus was assessed by stopwatch over a 1-month period. All men were genotyped for Cys23Ser 5-HT2c receptor gene polymorphism. Allele frequencies and genotypes of Cys and Ser variants of 5-HT2c receptor gene polymorphism were determined. Association between Cys/Cys and Ser/Ser genotypes and the natural logarithm of the IELT in men with LPE were.investigated. As a result, the geometric mean, median and natural mean IELT were 25.2, 27.0, 33.9s, respectively. Of all men, 20.0%, 10.8%, 23.1% and 41.5% ejaculated within 10, 10-20, 20-30 and 30-60s after vaginal penetration. Of the 64 men, the Cys/Cys and Ser/Ser genotype frequency for the Cys23Ser polymorphism of the 5-HT2c receptor gene was 81% and 19%, respectively. The geometric mean IELT of the wildtypes （Cys/Cys） is significantly lower （22.6s; 95% CI 18.3-27.8s） than in male homozygous mutants （Ser/Ser） （40.4s; 95% CI 20.3-80.4s） （P = 0.03）. It is concluded that Cys23Ser 5-HT2c receptor gene polymorphism is associated with the IELT in men with LPE. Men with Cys/Cys genotype have shorter IELTs than men with Ser/Ser genotypes.
