[Objective] The differential expression analysis was performed for FaGF14- B and FaGF14-C genes in tall fescue so as to provide certain basis for follow-up functional analysis of genes. [Method] The sequence fragments...[Objective] The differential expression analysis was performed for FaGF14- B and FaGF14-C genes in tall fescue so as to provide certain basis for follow-up functional analysis of genes. [Method] The sequence fragments of FaGF14-B and FaGF14-C obtained from reverse transcription were used as templates, and the full- length cDNA sequences of FaGF14-B and FaGF14-C were amplified using the 5＇ RACE use 3＇RACE techniques. They were named as FaGF14-B and FaGF14-C, and used for nucleic acid sequence analysis, encoded protein analysis, protein con- served domain analysis, phylogenetic analysis and differential expression analysis. [Result] The FaGF14-B gene has a full length of 1 548 bp. It has a complete open reading frame （ORF, 449-1 228 bp）, and encodes a protein composed of 261 amino acids. The FaGF14-C gene has a full length of 1 250 bp. It also has a complete open reading frame （ORF, 66-848 bp）, and encodes a protein composed of 261 amino acids. The GF14-B and GF14-C proteins all have a typical domain 14-3-3, and their secondary structures all contain 9 conserved co-helical structures and non-conserved N- and C- terminals. The phylogenetic analysis showed that the FaGF14-B and FaGF14-C from tall rescue have high similarities with GF14 protein from gramineous plants, and they are divided into the same clade with closer ge- netic relationship. The real-time fluorescence quantitative PCR analysis showed that the expression of FaGF14-B and FaGF14-C genes is all sensitive to nitrogen stress. [Conclusion] This study will lay a theoretical basis for further screening of low nitrogen-tolerant genes and breeding of low nitrogen-tolerant grass germplasms.展开更多
OBJECTIVE: To evaluate whether deletion of chromosome 14q is involved in the carcinogenesis of primary glioblastoma multiforme and to identify possibly common deletion regions. METHJODS: Fourteen fluorescent dye-label...OBJECTIVE: To evaluate whether deletion of chromosome 14q is involved in the carcinogenesis of primary glioblastoma multiforme and to identify possibly common deletion regions. METHJODS: Fourteen fluorescent dye-labeled polymorphic markers were used and polymerase chain reaction-based microsatellite analysis was employed to investigate loss of heterozygosity (LOH) on chromosome 14q in 20 primary glioblastoma multiforme (GBM). RESULTS: Ten of twenty (50%) GBM displayed LOH at one or more of the markers on chromosome 14q. Five tumors showed either LOH or non-informative on all markers tested. The most frequent LOH was observed at locus D14S65 (57.1%) on 14q32.1, and in the chromosomal region spanning from D14S63 (47.1%) to D14S74 (46.7%) on 14q23-31. None of the informative loci exhibited microsatellite instability. CONCLUSIONS: Allelic deletion on chromosome 14q plays an important role in the pathogenesis of GBM. Chromosomal regions at locus D14S65 on 14q32.1 and spanning from D14S63 to D14S74 on 14q23-31 may harbor multiple tumor suppressor genes associated with GBM.展开更多
Objective To investigate the relationship between alterations of p16 INK4a and p14 ARF genes and gastric carcinogenesis Methods The tumors and neighboring gastric tissues from 48 patients with gastric ca...Objective To investigate the relationship between alterations of p16 INK4a and p14 ARF genes and gastric carcinogenesis Methods The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16 INK4a and p14 ARF genes were assessed by PCR, PCR SSCP, PCR based methylation assay, and RT PCR Results ① The homozygous deletion rate of p16 INK4a and p14 ARF was 35 4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor ② There was no point mutation of p16 INK4a and p14 ARF in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor ③ Methylation of the CpG islands of p16 INK4a and p14 ARF was detected in 47 9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference ( P <0 01) ④ The loss rate of p16 INK4a mRNA was 47 9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1α and 2 had a higher loss rate (100%, 6/6) of p16 INK4a mRNA than those of the methylation of the other exons (11 8%, 2/17, P <0 01); the loss rate of p14 ARF mRNA was 45 8%(22/48) in gastric cancer, and patients with the combined methylation of exons 1β and 2 had a higher loss rate (100%, 3/3) of p14 ARF mRNA than those of the methylation of the other exons (15%, 3/20, P <0 05) ⑤ The combined loss of p16 INK4a and p14 ARF mRNAs was examined in 1 (5 6%) of 18 patients of well and moderately differentiated carcinomas, and 11 (36 7%) of 30 patients of poorly and not differentiated carcinomas with a significant difference ( P <0 05) Conclusion p16 INK4a and p14 ARF genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer展开更多
目的:双特异性酪氨酸磷酸化调节激酶,也称CDC样激酶(cell division cycle like kinases,CDC-like kinases,CLK),是一种双特异性蛋白激酶,其参与神经系统疾病、代谢异常、肿瘤等多个病理及生理过程,其中CLK3参与肝癌、乳腺癌等多种肿瘤...目的:双特异性酪氨酸磷酸化调节激酶,也称CDC样激酶(cell division cycle like kinases,CDC-like kinases,CLK),是一种双特异性蛋白激酶,其参与神经系统疾病、代谢异常、肿瘤等多个病理及生理过程,其中CLK3参与肝癌、乳腺癌等多种肿瘤的发生和发展,但其在结直肠癌中的表达情况及临床意义尚无相关报道。本研究通过挖掘肿瘤在线数据库,探讨CLK3在结直肠癌中的表达及临床意义。方法:利用基因表达谱交互分析2(Gene Expression Profiling Interactive Analysis 2,GEPIA2)数据库及GEO2R工具对CLK3在肿瘤间及结直肠癌组织和正常组织间作差异分析,获取差异表达结果后利用UALCN(The University of Alabama at Birmingham Cancer Data Analysis Portal)数据库对CLK3表达差异组作Kaplan-Meier生存分析。利用LinkedOmics数据库对CLK3进行共表达分析,并筛选出与之呈正相关及负相关的各50个基因,然后对获取的100个基因进行基因本体(gene ontology,GO)和京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,同时利用TIMER2.0数据库对CLK3进行免疫相关分析。在组织表达层面,利用人类蛋白质图谱(Human Protein Atlas,HPA)数据库查询CLK3在结直肠癌与正常肠道组织的免疫组织化学标本,分析其表达差异。结果:在UALCN数据库中,与正常组织比较,CLK3在结肠癌组织中低表达(P<0.01),而在直肠癌组织中表达差异无统计学意义(P>0.05)。CLK3的表达水平与结直肠癌患者的预后显著相关(P<0.05),表现为其高表达与不良预后相关。GO与KEGG通路富集结果显示:CLK3共表达基因富集于抗原肽呈递、物质代谢合成过程、转录调节过程的酶反应、γ干扰素介导的信号通路、Wnt信号通路及肠道免疫炎症过程等。相关性分析显示:结肠癌中CLK3表达与BRAF、HERS、NTRK1、PIK3CA基因突变相关,直肠癌中CLK3表达与BRAF和PIK3CA基因突变相关。进一步免疫浸润分析结果显示:在结肠癌中,CLK3的表达水平与CD8^(+)T细胞、CD4^(+)T细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05);在直肠癌中,CLK3的表达水平与CD4^(+)T细胞、巨噬细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05)。在免疫组织化学染色切片中,结肠癌和直肠癌CLK3蛋白表达水平均低于正常结肠组织(均P<0.05)。结论:CLK3在结直肠癌中存在差异表达,是结直肠癌生存预后的指标,有望成为结直肠癌的预后评估和临床治疗的有效靶点之一。展开更多
基金Supported by National Natural Science Foundation of China(31360576)~~
文摘[Objective] The differential expression analysis was performed for FaGF14- B and FaGF14-C genes in tall fescue so as to provide certain basis for follow-up functional analysis of genes. [Method] The sequence fragments of FaGF14-B and FaGF14-C obtained from reverse transcription were used as templates, and the full- length cDNA sequences of FaGF14-B and FaGF14-C were amplified using the 5＇ RACE use 3＇RACE techniques. They were named as FaGF14-B and FaGF14-C, and used for nucleic acid sequence analysis, encoded protein analysis, protein con- served domain analysis, phylogenetic analysis and differential expression analysis. [Result] The FaGF14-B gene has a full length of 1 548 bp. It has a complete open reading frame （ORF, 449-1 228 bp）, and encodes a protein composed of 261 amino acids. The FaGF14-C gene has a full length of 1 250 bp. It also has a complete open reading frame （ORF, 66-848 bp）, and encodes a protein composed of 261 amino acids. The GF14-B and GF14-C proteins all have a typical domain 14-3-3, and their secondary structures all contain 9 conserved co-helical structures and non-conserved N- and C- terminals. The phylogenetic analysis showed that the FaGF14-B and FaGF14-C from tall rescue have high similarities with GF14 protein from gramineous plants, and they are divided into the same clade with closer ge- netic relationship. The real-time fluorescence quantitative PCR analysis showed that the expression of FaGF14-B and FaGF14-C genes is all sensitive to nitrogen stress. [Conclusion] This study will lay a theoretical basis for further screening of low nitrogen-tolerant genes and breeding of low nitrogen-tolerant grass germplasms.
文摘OBJECTIVE: To evaluate whether deletion of chromosome 14q is involved in the carcinogenesis of primary glioblastoma multiforme and to identify possibly common deletion regions. METHJODS: Fourteen fluorescent dye-labeled polymorphic markers were used and polymerase chain reaction-based microsatellite analysis was employed to investigate loss of heterozygosity (LOH) on chromosome 14q in 20 primary glioblastoma multiforme (GBM). RESULTS: Ten of twenty (50%) GBM displayed LOH at one or more of the markers on chromosome 14q. Five tumors showed either LOH or non-informative on all markers tested. The most frequent LOH was observed at locus D14S65 (57.1%) on 14q32.1, and in the chromosomal region spanning from D14S63 (47.1%) to D14S74 (46.7%) on 14q23-31. None of the informative loci exhibited microsatellite instability. CONCLUSIONS: Allelic deletion on chromosome 14q plays an important role in the pathogenesis of GBM. Chromosomal regions at locus D14S65 on 14q32.1 and spanning from D14S63 to D14S74 on 14q23-31 may harbor multiple tumor suppressor genes associated with GBM.
文摘Objective To investigate the relationship between alterations of p16 INK4a and p14 ARF genes and gastric carcinogenesis Methods The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16 INK4a and p14 ARF genes were assessed by PCR, PCR SSCP, PCR based methylation assay, and RT PCR Results ① The homozygous deletion rate of p16 INK4a and p14 ARF was 35 4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor ② There was no point mutation of p16 INK4a and p14 ARF in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor ③ Methylation of the CpG islands of p16 INK4a and p14 ARF was detected in 47 9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference ( P <0 01) ④ The loss rate of p16 INK4a mRNA was 47 9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1α and 2 had a higher loss rate (100%, 6/6) of p16 INK4a mRNA than those of the methylation of the other exons (11 8%, 2/17, P <0 01); the loss rate of p14 ARF mRNA was 45 8%(22/48) in gastric cancer, and patients with the combined methylation of exons 1β and 2 had a higher loss rate (100%, 3/3) of p14 ARF mRNA than those of the methylation of the other exons (15%, 3/20, P <0 05) ⑤ The combined loss of p16 INK4a and p14 ARF mRNAs was examined in 1 (5 6%) of 18 patients of well and moderately differentiated carcinomas, and 11 (36 7%) of 30 patients of poorly and not differentiated carcinomas with a significant difference ( P <0 05) Conclusion p16 INK4a and p14 ARF genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer
文摘目的:双特异性酪氨酸磷酸化调节激酶,也称CDC样激酶(cell division cycle like kinases,CDC-like kinases,CLK),是一种双特异性蛋白激酶,其参与神经系统疾病、代谢异常、肿瘤等多个病理及生理过程,其中CLK3参与肝癌、乳腺癌等多种肿瘤的发生和发展,但其在结直肠癌中的表达情况及临床意义尚无相关报道。本研究通过挖掘肿瘤在线数据库,探讨CLK3在结直肠癌中的表达及临床意义。方法:利用基因表达谱交互分析2(Gene Expression Profiling Interactive Analysis 2,GEPIA2)数据库及GEO2R工具对CLK3在肿瘤间及结直肠癌组织和正常组织间作差异分析,获取差异表达结果后利用UALCN(The University of Alabama at Birmingham Cancer Data Analysis Portal)数据库对CLK3表达差异组作Kaplan-Meier生存分析。利用LinkedOmics数据库对CLK3进行共表达分析,并筛选出与之呈正相关及负相关的各50个基因,然后对获取的100个基因进行基因本体(gene ontology,GO)和京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,同时利用TIMER2.0数据库对CLK3进行免疫相关分析。在组织表达层面,利用人类蛋白质图谱(Human Protein Atlas,HPA)数据库查询CLK3在结直肠癌与正常肠道组织的免疫组织化学标本,分析其表达差异。结果:在UALCN数据库中,与正常组织比较,CLK3在结肠癌组织中低表达(P<0.01),而在直肠癌组织中表达差异无统计学意义(P>0.05)。CLK3的表达水平与结直肠癌患者的预后显著相关(P<0.05),表现为其高表达与不良预后相关。GO与KEGG通路富集结果显示:CLK3共表达基因富集于抗原肽呈递、物质代谢合成过程、转录调节过程的酶反应、γ干扰素介导的信号通路、Wnt信号通路及肠道免疫炎症过程等。相关性分析显示:结肠癌中CLK3表达与BRAF、HERS、NTRK1、PIK3CA基因突变相关,直肠癌中CLK3表达与BRAF和PIK3CA基因突变相关。进一步免疫浸润分析结果显示:在结肠癌中,CLK3的表达水平与CD8^(+)T细胞、CD4^(+)T细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05);在直肠癌中,CLK3的表达水平与CD4^(+)T细胞、巨噬细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05)。在免疫组织化学染色切片中,结肠癌和直肠癌CLK3蛋白表达水平均低于正常结肠组织(均P<0.05)。结论:CLK3在结直肠癌中存在差异表达,是结直肠癌生存预后的指标,有望成为结直肠癌的预后评估和临床治疗的有效靶点之一。
