Objective： To screen and analyze key express sequence tags （ESTs） which were differentially displayed in every period of SD rats＇ primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Met...Objective： To screen and analyze key express sequence tags （ESTs） which were differentially displayed in every period of SD rats＇ primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Methods： Using diethylnitrosamine （DENA） as a cancerigenic agent, animal models with different phases of primary hepatic cancer were constructed in SD rats. Rats were respectively sacrificed at d 14, d 28, d 56, d 77, d 105 and d 112 after the rats received DENA by gavage, then the livers were harvested. One part of the livers was classified according to their pathological changes, while the other was reserved for molecular mechanism studies on hepatocarcinogenesis. The differentially expressed genes were isolated from both normal and morbid tissues by mRNA differential display technique （DDRT-PCR）. After the fragments were sequenced, bioinformatics were .used to analyze the results. Results： Twelve differentially expressed cDNA fragments were obtained. Nine fragments had the homology with known cDNA clones, especially EST-7 was similar to BN/SsNHsdMCW mitochondrion gene and the identity was 100% which suggested EST-7 may be the part of BN/SsNHsdMCW mitochondrion gene. In contrast, other three fragments （EST-1, EST-3 and EST-5） had extremely low identity to any genes registered in GENBANK databases. Conclusions： BN/SsNHsdMCW mitochondrion gene was expressed in different periods of hepatocarcinogenesis. Moreover, EST-I, EST-3 and EST-5 were suggested to contribute to the development of rat hepatocarcinogenesis, and thus may be candidates of new targets of oncogenes or cancer suppressor genes.展开更多
AIM： To investigate the differently expressed genes in human colorectal adenocarcinorna.METHODS： The integrated approach for gene expression profiling that couples suppression subtractive hybridization, high-through...AIM： To investigate the differently expressed genes in human colorectal adenocarcinorna.METHODS： The integrated approach for gene expression profiling that couples suppression subtractive hybridization, high-throughput cDNA array, sequencing, bioinformatics analysis, and reverse transcriptase real- time quantitative polymerase chain reaction （PCR） was carried out. A set of cDNA clones including 1260 SSH inserts amplified by PCR was arrayed using robotic printing. The cDNA arrays were hybridized with florescent-labeled probes prepared from RNA of human colorectal adenocarcinoma （HCRAC） and normal colorectal tissues.RESULTS： A total of 86 genes were identified, 16 unknown genes and 70 known genes. The transcription factor Sox9 influencing cell differentiation was downregulated. At the same time, Heat shock protein 10 KDis downregulated and Calmoulin is up-regulated.CONCLUSION： Downregulation of heat shock protein 10 KD lost its inhibition of Ras, and men attenuated the Ras GTPase signaling pathway, increased cell proliferation and inhibited cell apoptosis. Down-regulated transcription factor Sox9 influences cell differentiation and cell-specific gene expression. Down-regulated Sox9 also decreases its binding to calmodulin, accumulates calmodulin as receptor-activated kinase and phosphorylase kinase due to the activation of PhK.展开更多
The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse tran...The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.展开更多
Chemical inducement and DDRT-PCR （differential display reverse transcription PCR） are adopted to investigate the effect of epinephrine （EPI） on the settlement and metamorphosis of Manila clam larvae. Chemical indu...Chemical inducement and DDRT-PCR （differential display reverse transcription PCR） are adopted to investigate the effect of epinephrine （EPI） on the settlement and metamorphosis of Manila clam larvae. Chemical inducement shows that EPI has an effect to some extent on the metamorphosis of Manila clam larvae at all concentrations and in all treatments designed. The most significant result of inducement is obtained at the concentration of 10^-6 tool L^-1 and for 4 h. DDRT-PCR using six primer pairs shows that the gene expression pattern is quite different between EPI treatment and the control. Three hundred and forty-three amplification bands are obtained in total, among which, 67 （19.53%） are differentially appeared. Therefore, EPI has an effect on the gene expression of the eye spot larval Manila clam. It can be hypothesized that EPI is a settlement and metamorphosis inducer for Manila clam. EPI may lead to larvae settlement and metamorphosis by binding to the receptors on the membrane and then changing the gene expression of larvae cells.展开更多
Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study a...Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study aimed to explore the effect of ITIH4 on OC and to identify its underlying mechanism.Methods:Expressions of ITIH4 in OC tissues and cells were determined using quantitative reverse transcription polymerase chain reaction(RT-qPCR)and western blots.The function of ITIH4 in the OC cell line HO8910 pm was tested via ITIH4 knockdown.The cell growth rate was measured using MTT and colony formation assays.Flow cytometry was performed to evaluate cell cycle progression.Cell migration and invasion abilities were observed using the transwell migration assay.Results:ITIH4 was downregulated in OC tissues and cells.ITIH4 knockdown promoted cell growth and cell cycle progression.Consistent with these results,inhibition of ITIH4 in OC cells significantly increased cell migration and invasion abilities.Cox regression analysis suggests that ITIH4 expression alone is not a good predictor of the prognosis of malignant ovarian tumors in patients.Conclusions:ITIH4 inhibits the progression of OC,suggesting that ITIH4 may be a useful biomarker for OC.This study may provide a potential novel target for the treatment of OC.展开更多
Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand ...Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand molecular mechanisms of root and stem rot resistance in soybeans, the gene and protein expression in hypocotyls and stems of variety Suinong 10 carrying resistance genes Rps1a and Rps2 was investigated by using mRNA differential display reverse transcription PCR and two-dimensional electrophoresis at 0, 0.5, 1, 2, and 4 h after inoculation with P. sojae race 1. The results of the comparison of gene and protein expression showed that at least eight differential fragments at the transcriptional level were related to metabolic pathway, phytoalexin, and signal transduction in defense responses. Sequence analyses indicated that these fragments represented cinnamic acid 4-hydroxylase gene, ATP b gene coding ATP synthase b subunit and ubiquitin-conjugating enzyme gene which upregulated at 0.5 h post inoculation, blue copper protein gene and UDP-N-acetyl-a-D-galactosamine gene which upregulated at 2 h post inoculation, TGA-type basic leucine zipper protein TGA1.1 gene, cyclophilin gene, and 14-3-3 protein gene which upregulated at 4 h post inoculation. Three resistance-related proteins, a-subunit and b-subunit of ATP synthase, and cytochrome P450-like protein, were upregulated at 2 h post inoculation. The results suggested that resistance-related multiple proteins and genes were expressed in the recognition between soybean and P. sojae during zoospore germination, penetration and mycelium growth of P. sojae in soybean.展开更多
Objective： To investigate the expression of maspin, uPA and MMP-7 in gastric carcinoma and discuss their roles in tumorigenesis and progression of gastric cancer. Methods： lmmunohistochemistry SP method was used to ...Objective： To investigate the expression of maspin, uPA and MMP-7 in gastric carcinoma and discuss their roles in tumorigenesis and progression of gastric cancer. Methods： lmmunohistochemistry SP method was used to detect expression of maspin, uPA and MMP-7 in gastric adenocarcinoma （n=30）, signet-ring cell carcinoma （n=30）, normal gastric mucosa （n=20）; in addition, reverse transcriptase PCR was carried out to compare the mRNA expression of maspin between gastric adenocarcinoma and normal mucosa. Results： The expression rates of maspin, uPA and MMP-7 protein were 50%, 70% and 80% in gastric adenocarcinoma, and 46.7%, 76.7%, and 90% in signet-ring cell carcinoma, respectively. However, the expression rates of maspin, uPA, and MMP-7 protein were 90%, 35% and 30%, respectively in normal gastric mucosa. The relative transcription level of maspin mRNA in normal gastric mucosa was higher than that in gastric adenocarcinoma （P〈0.01）. Maspin protein expression was significantly related to the depth of invasion and lymph node metastasis of gastric carcinoma, but did not relate to tumor size and TNM staging. The protein expression of uPA and MMP-7 were significantly related to invasive depth, lymph node metastasis and TNM staging of carcinoma, but not to tumor size. Maspin protein expression had a negative correlation with uPA and MMP-7, while uPA protein expression had a positive correlation with MMP-7. The relative mRNA transcription level of Maspin significantly correlated with the lymph node metastasis and the depth of invasion of carcinoma, but did not relate to TNM staging or tumor size. Conclusion： Down-regulated expression of maspin and up- regulated expression of uPA and MMP-7 play important roles in the invasion and metastasis of gastric carcinoma. They may serve as effective markers of the biopathological behavior of gastric tumor.展开更多
The heptadecapeptide orphanin FQ (OFQ) is a recently discovered neuropeptide that exhibits structural features reminiscent of the opioid peptides and that is an endogenous ligant to a G protein-coupled receptor sequen...The heptadecapeptide orphanin FQ (OFQ) is a recently discovered neuropeptide that exhibits structural features reminiscent of the opioid peptides and that is an endogenous ligant to a G protein-coupled receptor sequentially related to the opioid receptors. OFQ was originally isolated from brain, but the presence of OFQ in peripheral tissues, especially in cardiovascular system, has not been clarified. The present study was designed to investigate the peripheral tissue distribution of OFQ precusor mRNA in stroke-prone spontaneously hypertensive rats (SHRSP) and compare the difference of OFQ precusor mRNA expression in aorta or cultured vascular smooth muscle cells (VSMCs) between SHRSP and wistar-Kyoto normotensive (WKY) rats. By using quantitative reverse transcription-polymerase chain reaction (RT-PCR), OFQ precusor mRNA was detected in aorta and ovary at high levels comparable with the amounts found in brain. Moderate expression was found in testis, while a little OFQ precusor mRNA could be detected in atrium. All other peripheral tissues examined from SHRSP, including ventricle, liver, lung and kidney, showed no expression of OFQ precusor mRNA. In the vascular system, OFQ precusor mRNA was expressed in aorta, pulmonary artery, renal artery and vein at high levels comparable with the amounts found in brain. We also found that OFQ precusor mRNA levels were much higher in aorta or cultured VSMCs from SHRSP than those from WKY rats. In conclusion, the present study has shown that OFQ precusor mRNA is present in some peripheral tissues, especially in cardiovascular and reproductive system, suggesting that OFQ possibly involves in the regulation of cardiovascular and reproductive functions.展开更多
Although gonadotropin releasing hormone (GnRH), GnRH like molecule, and GnRH receptor (GnRH R) have been reported to exist in several tissues other than brain or anterior pituitary, there are no reports concerning ...Although gonadotropin releasing hormone (GnRH), GnRH like molecule, and GnRH receptor (GnRH R) have been reported to exist in several tissues other than brain or anterior pituitary, there are no reports concerning GnRH or GnRH R gene expression in a normal pancreatic gland. In order to define the production of GnRH as well as GnRH R in the pancreatic gland, we examined their gene expression in various developmental stages of rat pancreas using the reverse transcriptase polymerase chain reaction (RT PCR). GnRH mRNA transcripts were found in pancreas of male and female rats at different ages, expressing at about the same level, whereas GnRH R mRNA transcripts could not be detected in any rat pancreatic gland samples. These results suggest a possible biological role of GnRH in rodent pancreas.展开更多
基金supported by the Key Program for Science and Technology Development of Henan Province [122102310174]the Zoology Key Subject of Henan Province
文摘Objective： To screen and analyze key express sequence tags （ESTs） which were differentially displayed in every period of SD rats＇ primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Methods： Using diethylnitrosamine （DENA） as a cancerigenic agent, animal models with different phases of primary hepatic cancer were constructed in SD rats. Rats were respectively sacrificed at d 14, d 28, d 56, d 77, d 105 and d 112 after the rats received DENA by gavage, then the livers were harvested. One part of the livers was classified according to their pathological changes, while the other was reserved for molecular mechanism studies on hepatocarcinogenesis. The differentially expressed genes were isolated from both normal and morbid tissues by mRNA differential display technique （DDRT-PCR）. After the fragments were sequenced, bioinformatics were .used to analyze the results. Results： Twelve differentially expressed cDNA fragments were obtained. Nine fragments had the homology with known cDNA clones, especially EST-7 was similar to BN/SsNHsdMCW mitochondrion gene and the identity was 100% which suggested EST-7 may be the part of BN/SsNHsdMCW mitochondrion gene. In contrast, other three fragments （EST-1, EST-3 and EST-5） had extremely low identity to any genes registered in GENBANK databases. Conclusions： BN/SsNHsdMCW mitochondrion gene was expressed in different periods of hepatocarcinogenesis. Moreover, EST-I, EST-3 and EST-5 were suggested to contribute to the development of rat hepatocarcinogenesis, and thus may be candidates of new targets of oncogenes or cancer suppressor genes.
基金Supported by postdoctoral fellowship from China National Human Affairs Ministry to Chen Yao,Development Program of Sichuan University to Chen YaoChina National Excellent Youth Fund to Zhong-Guang Zhou,No.39925032
文摘AIM： To investigate the differently expressed genes in human colorectal adenocarcinorna.METHODS： The integrated approach for gene expression profiling that couples suppression subtractive hybridization, high-throughput cDNA array, sequencing, bioinformatics analysis, and reverse transcriptase real- time quantitative polymerase chain reaction （PCR） was carried out. A set of cDNA clones including 1260 SSH inserts amplified by PCR was arrayed using robotic printing. The cDNA arrays were hybridized with florescent-labeled probes prepared from RNA of human colorectal adenocarcinoma （HCRAC） and normal colorectal tissues.RESULTS： A total of 86 genes were identified, 16 unknown genes and 70 known genes. The transcription factor Sox9 influencing cell differentiation was downregulated. At the same time, Heat shock protein 10 KDis downregulated and Calmoulin is up-regulated.CONCLUSION： Downregulation of heat shock protein 10 KD lost its inhibition of Ras, and men attenuated the Ras GTPase signaling pathway, increased cell proliferation and inhibited cell apoptosis. Down-regulated transcription factor Sox9 influences cell differentiation and cell-specific gene expression. Down-regulated Sox9 also decreases its binding to calmodulin, accumulates calmodulin as receptor-activated kinase and phosphorylase kinase due to the activation of PhK.
基金This work was financially supported by the National Natural Science Foundation of China （30170682, 30571400） the Specialized Research Fund for the Doctoral Program of Higher Education, China （20050307008）.
文摘The relationship between myocardial cell damages and HSPs mRNA transcription in heat stressed broilers was studied using a spectrophotometer, the histopathological technique, and fluorescence quantitative reverse transcription PCR （FQ RT-PCR）. The results showed that the activities of creatine kinase （CK） and glutamic-pyruvic transaminase （GPT） were induction during the persistent heat stress. The major lesions of the myocardial fibers were granular degeneration and necrosis. The transcription of constitutive or cognate heat shock protein 70 （HSC70） mRNA was changeable. The transcription of heat shock protein 70 （HSPT0） mRNA was increased obviously in the course of persistent heat stress. The results showed that the change of HSC70 mRNA transcription was contrary to the activity of CK, and the level of HSC70 mRNA transcription must be used as a symbol of the myocardial cell damages in the course of persistent heat stress.
文摘Chemical inducement and DDRT-PCR （differential display reverse transcription PCR） are adopted to investigate the effect of epinephrine （EPI） on the settlement and metamorphosis of Manila clam larvae. Chemical inducement shows that EPI has an effect to some extent on the metamorphosis of Manila clam larvae at all concentrations and in all treatments designed. The most significant result of inducement is obtained at the concentration of 10^-6 tool L^-1 and for 4 h. DDRT-PCR using six primer pairs shows that the gene expression pattern is quite different between EPI treatment and the control. Three hundred and forty-three amplification bands are obtained in total, among which, 67 （19.53%） are differentially appeared. Therefore, EPI has an effect on the gene expression of the eye spot larval Manila clam. It can be hypothesized that EPI is a settlement and metamorphosis inducer for Manila clam. EPI may lead to larvae settlement and metamorphosis by binding to the receptors on the membrane and then changing the gene expression of larvae cells.
基金supported by Guangxi Scientific Research and Technology Development Project (No. 14124004-1-24)
文摘Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study aimed to explore the effect of ITIH4 on OC and to identify its underlying mechanism.Methods:Expressions of ITIH4 in OC tissues and cells were determined using quantitative reverse transcription polymerase chain reaction(RT-qPCR)and western blots.The function of ITIH4 in the OC cell line HO8910 pm was tested via ITIH4 knockdown.The cell growth rate was measured using MTT and colony formation assays.Flow cytometry was performed to evaluate cell cycle progression.Cell migration and invasion abilities were observed using the transwell migration assay.Results:ITIH4 was downregulated in OC tissues and cells.ITIH4 knockdown promoted cell growth and cell cycle progression.Consistent with these results,inhibition of ITIH4 in OC cells significantly increased cell migration and invasion abilities.Cox regression analysis suggests that ITIH4 expression alone is not a good predictor of the prognosis of malignant ovarian tumors in patients.Conclusions:ITIH4 inhibits the progression of OC,suggesting that ITIH4 may be a useful biomarker for OC.This study may provide a potential novel target for the treatment of OC.
基金supported by the Commonweal Specialized Research Fund of China Agriculture (3-20,201103015)
文摘Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand molecular mechanisms of root and stem rot resistance in soybeans, the gene and protein expression in hypocotyls and stems of variety Suinong 10 carrying resistance genes Rps1a and Rps2 was investigated by using mRNA differential display reverse transcription PCR and two-dimensional electrophoresis at 0, 0.5, 1, 2, and 4 h after inoculation with P. sojae race 1. The results of the comparison of gene and protein expression showed that at least eight differential fragments at the transcriptional level were related to metabolic pathway, phytoalexin, and signal transduction in defense responses. Sequence analyses indicated that these fragments represented cinnamic acid 4-hydroxylase gene, ATP b gene coding ATP synthase b subunit and ubiquitin-conjugating enzyme gene which upregulated at 0.5 h post inoculation, blue copper protein gene and UDP-N-acetyl-a-D-galactosamine gene which upregulated at 2 h post inoculation, TGA-type basic leucine zipper protein TGA1.1 gene, cyclophilin gene, and 14-3-3 protein gene which upregulated at 4 h post inoculation. Three resistance-related proteins, a-subunit and b-subunit of ATP synthase, and cytochrome P450-like protein, were upregulated at 2 h post inoculation. The results suggested that resistance-related multiple proteins and genes were expressed in the recognition between soybean and P. sojae during zoospore germination, penetration and mycelium growth of P. sojae in soybean.
基金a grant from the project of Chongqing Educational Committee(No.040310)
文摘Objective： To investigate the expression of maspin, uPA and MMP-7 in gastric carcinoma and discuss their roles in tumorigenesis and progression of gastric cancer. Methods： lmmunohistochemistry SP method was used to detect expression of maspin, uPA and MMP-7 in gastric adenocarcinoma （n=30）, signet-ring cell carcinoma （n=30）, normal gastric mucosa （n=20）; in addition, reverse transcriptase PCR was carried out to compare the mRNA expression of maspin between gastric adenocarcinoma and normal mucosa. Results： The expression rates of maspin, uPA and MMP-7 protein were 50%, 70% and 80% in gastric adenocarcinoma, and 46.7%, 76.7%, and 90% in signet-ring cell carcinoma, respectively. However, the expression rates of maspin, uPA, and MMP-7 protein were 90%, 35% and 30%, respectively in normal gastric mucosa. The relative transcription level of maspin mRNA in normal gastric mucosa was higher than that in gastric adenocarcinoma （P〈0.01）. Maspin protein expression was significantly related to the depth of invasion and lymph node metastasis of gastric carcinoma, but did not relate to tumor size and TNM staging. The protein expression of uPA and MMP-7 were significantly related to invasive depth, lymph node metastasis and TNM staging of carcinoma, but not to tumor size. Maspin protein expression had a negative correlation with uPA and MMP-7, while uPA protein expression had a positive correlation with MMP-7. The relative mRNA transcription level of Maspin significantly correlated with the lymph node metastasis and the depth of invasion of carcinoma, but did not relate to TNM staging or tumor size. Conclusion： Down-regulated expression of maspin and up- regulated expression of uPA and MMP-7 play important roles in the invasion and metastasis of gastric carcinoma. They may serve as effective markers of the biopathological behavior of gastric tumor.
文摘The heptadecapeptide orphanin FQ (OFQ) is a recently discovered neuropeptide that exhibits structural features reminiscent of the opioid peptides and that is an endogenous ligant to a G protein-coupled receptor sequentially related to the opioid receptors. OFQ was originally isolated from brain, but the presence of OFQ in peripheral tissues, especially in cardiovascular system, has not been clarified. The present study was designed to investigate the peripheral tissue distribution of OFQ precusor mRNA in stroke-prone spontaneously hypertensive rats (SHRSP) and compare the difference of OFQ precusor mRNA expression in aorta or cultured vascular smooth muscle cells (VSMCs) between SHRSP and wistar-Kyoto normotensive (WKY) rats. By using quantitative reverse transcription-polymerase chain reaction (RT-PCR), OFQ precusor mRNA was detected in aorta and ovary at high levels comparable with the amounts found in brain. Moderate expression was found in testis, while a little OFQ precusor mRNA could be detected in atrium. All other peripheral tissues examined from SHRSP, including ventricle, liver, lung and kidney, showed no expression of OFQ precusor mRNA. In the vascular system, OFQ precusor mRNA was expressed in aorta, pulmonary artery, renal artery and vein at high levels comparable with the amounts found in brain. We also found that OFQ precusor mRNA levels were much higher in aorta or cultured VSMCs from SHRSP than those from WKY rats. In conclusion, the present study has shown that OFQ precusor mRNA is present in some peripheral tissues, especially in cardiovascular and reproductive system, suggesting that OFQ possibly involves in the regulation of cardiovascular and reproductive functions.
基金the National Natural Sciences Foundationof China(No. 39770 388)
文摘Although gonadotropin releasing hormone (GnRH), GnRH like molecule, and GnRH receptor (GnRH R) have been reported to exist in several tissues other than brain or anterior pituitary, there are no reports concerning GnRH or GnRH R gene expression in a normal pancreatic gland. In order to define the production of GnRH as well as GnRH R in the pancreatic gland, we examined their gene expression in various developmental stages of rat pancreas using the reverse transcriptase polymerase chain reaction (RT PCR). GnRH mRNA transcripts were found in pancreas of male and female rats at different ages, expressing at about the same level, whereas GnRH R mRNA transcripts could not be detected in any rat pancreatic gland samples. These results suggest a possible biological role of GnRH in rodent pancreas.
