AIM: To comprehensively understand the underlying molecular events accounting for aberrant Wnt signaling activation in hepatocellular carcinoma(HCC).METHODS: This study was retrospective. The HCC tissue specimens used...AIM: To comprehensively understand the underlying molecular events accounting for aberrant Wnt signaling activation in hepatocellular carcinoma(HCC).METHODS: This study was retrospective. The HCC tissue specimens used in this research were obtained from patients who underwent liver surgery. The Catalogue of Somatic Mutations in Cancer(COSMIC) database was searched for the mutation statuses of CTNNB1, TP53, and protein degradation regulator genes of CTNNB1. Dual-luciferase reporter assay was performed with TOP/FOP reporters to detect whether TP53 gain-of-function(GOF) mutations could enhance the transcriptional activity of Wnt signaling. Methylation sensitive restriction enzyme-quantitative PCR was used to explore the methylation status of Cp G islands located in the promoters of APC, SFRP1, and SFRP5 in HCCs with different risk factors. Finally, nestedreverse transcription PCR was performed to examine the integration of HBx in front of LINE1 element and the existence of HBx-LINE1 chimeric transcript in Hepatitis B virus-related HCC. All results in this article were analyzed with the software SPSS version 19.0 for Windows, and different groups were compared by χ2 test as appropriate.RESULTS: Based on the data from COSMIC database, compared with other solid tumors, mutation frequency of CTNNB1 was significantly higher in HCC(P < 0.01). The rate of CTNNB1 mutation was significantly less frequent in Hepatitis B virus-related HCC than in other etiologies(P < 0.01). Dual-luciferase reporter system and TOP/FOP reporter assays confirmed that TP53 GOF mutants were able to enhance the transcriptional ability of Wnt signaling. An exclusive relationship between the status of TP53 and CTNNB1 mutations was observed. However, according to the COSMIC database, TP53 GOF mutation is rare in HCC, which indicates that TP53 GOF mutation is not a reason for the aberrant activation of Wnt signaling in HCC. APC and AXIN1 were mutated in HCC. By using methylation sensitive restriction enzyme-quantitative PCR, hypermethylation of APC was展开更多
基于整合药理学和实验验证探讨健脾除痰解毒方(Jianpi Chutan Jiedu Decoction,JCJD)干预肺癌的作用机制。采用中医药整合药理学研究平台(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine,TCMIP)V2...基于整合药理学和实验验证探讨健脾除痰解毒方(Jianpi Chutan Jiedu Decoction,JCJD)干预肺癌的作用机制。采用中医药整合药理学研究平台(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine,TCMIP)V2.0,检索并获取健脾除痰解毒方靶标及功能信息,以及肺癌疾病靶标及功能信息,借助中医关联网络挖掘平台构建“疾病-方剂”关联网络,并通过网络拓扑特征计算、自定义多维关联网络可视化(方剂-中药材-核心靶标-通路-疾病)以及基因本体(GO)功能和依托Reactome数据库的Pathway信息进行富集分析,发现健脾除痰解毒方干预肺癌的关键网络靶标及其作用机制。最后通过动物实验进一步验证整合药理学结果。结果共获得健脾除痰解毒方干预肺癌的核心靶标53个;GO富集分析得到生物过程有20种条目,细胞组分有11种条目,分子功能有20种条目;Reactome Pathway富集分析得到11个条目,主要涉及呼吸电子传递、TP53调节代谢基因、细胞内受体的SUMO化、白细胞介素(IL)-4和IL-13通路等信号通路。动物实验结果显示与模型组比较,联合组肿瘤组织中乳酸含量明显下降(P<0.05),联合组可明显上调肿瘤组织中磷酸酯酶与张力蛋白同源物(PTEN)蛋白平均光密度值(P<0.05),降低丙酮酸激酶M2(PKM2)和己糖激酶2(HK2)蛋白平均光密度值(P<0.05);与模型组比较,联合组可明显上调肿瘤组织中肿瘤抑制蛋白(P53)、PTEN mRNA及蛋白表达(P<0.01,P<0.05),降低肿瘤组织中TP53诱导的糖酵解与凋亡调控因子(TIGAR)、葡萄糖转运蛋白1(GLUT1)、PKM2、HK2 mRNA及蛋白表达(P<0.01)。综上,健脾除痰解毒方通过TP53调节代谢基因来下调葡萄糖代谢,从而抑制肺癌增殖。展开更多
AIM:To rapidly detect molecular alterations in different malignancies and investigate the possible role of Tp53,C-myc,and CCND1 genes in development of tumors in human organs and their adjacent normal tissues,as well ...AIM:To rapidly detect molecular alterations in different malignancies and investigate the possible role of Tp53,C-myc,and CCND1 genes in development of tumors in human organs and their adjacent normal tissues,as well as the possible relation between well-and poorly-differentiated tumors.METHODS:A tissue array consisting of seven different tumors was generated.The tissue array included 120 points of esophagus,120 points of stomach,80 points of rectum,60 points of thyroid gland,100 points of mammary gland,80 points of liver,and 80 points of colon.Expressions of Tp53,C-myc,and CCND1 were determined by RNA in situ hybridization.3' terminal digoxin-labeled anti-sense single stranded oligonucleotide and locked nucleic acid modifying probe were used.RESULTS:The expression level of Tp53 gene was higher in six different carcinoma tissue samples than in paracancerous tissue samples with the exception in colon carcinoma tissue samples(P < 0.05).The expression level of CCND1 gene was significantly different in different carcinoma tissue samples with the exception in esophagus and colon carcinoma tissue samples.The expression level of C-myc gene was different in esophagus carcinoma tissue samples(c2 = 18.495,P = 0.000),stomach carcinoma tissue samples(c2 = 23.750,P = 0.000),and thyroid gland tissue samples(c2 = 10.999,P = 0.004).The intensity of signals was also different in different carcinoma tissue samples and paracancerous tissue samples.CONCLUSION:Over-expression of the Tp53,CCND1,and C-myc genes appears to play a role in development of human cancer by regulating the expression of mRNA.Tp53,CCND1 and C-myc genes are significantly correlated with the development of different carcinomas.展开更多
Tp53, a stress response gene, is involved in diverse cell death pathways and its activation is implicated in the pathogenesis of Parkinson＇s disease. However, whether the neuronal Tp53 protein plays a direct role in ...Tp53, a stress response gene, is involved in diverse cell death pathways and its activation is implicated in the pathogenesis of Parkinson＇s disease. However, whether the neuronal Tp53 protein plays a direct role in regulating dopaminergic （DA） neuronal cell death or neuronal terminal damage in different neurotoxicant models is unknown. In our recent studies, in contrast to the global inhibition of Tp53 function by phar- macological inhibitors and in traditional Tp53 knock-out mice, we examined the effects of DA-specific Tp53 gene deletion after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and methamphetamine exposure. Our data suggests that the Tp53 gene might be involved in both neuronal apoptosis and neuronal termi- nal damage caused by different neurotoxicants. Additional results from other studies also suggest that as a master regulator of many pathways that regulate apoptosis and synaptic terminal damage, it is possible that Tp53 may function as a signaling hub to integrate different signaling pathways to mediate distinctive target pathways. Tp53 protein as a signaling hub might be able to evaluate the microenvironment of neurons, assess the forms and severities of injury incurred, and determine whether apoptotic cell death or neuro- nal terminal degeneration occurs. Identification of the precise mechanisms activated in distinct neuronal damage caused by different forms and severities of injuries might allow for development of specific Tp53 inhibitors or ways to modulate distinct downstream target pathways involved.展开更多
基金Supported by National Natural Science Foundation of China,No.81372603973 Program,No.2015CB554000+1 种基金National S T Major Project for Infectious Diseases,No.2012ZX10004-904The 111 Project,No.B07001
文摘AIM: To comprehensively understand the underlying molecular events accounting for aberrant Wnt signaling activation in hepatocellular carcinoma(HCC).METHODS: This study was retrospective. The HCC tissue specimens used in this research were obtained from patients who underwent liver surgery. The Catalogue of Somatic Mutations in Cancer(COSMIC) database was searched for the mutation statuses of CTNNB1, TP53, and protein degradation regulator genes of CTNNB1. Dual-luciferase reporter assay was performed with TOP/FOP reporters to detect whether TP53 gain-of-function(GOF) mutations could enhance the transcriptional activity of Wnt signaling. Methylation sensitive restriction enzyme-quantitative PCR was used to explore the methylation status of Cp G islands located in the promoters of APC, SFRP1, and SFRP5 in HCCs with different risk factors. Finally, nestedreverse transcription PCR was performed to examine the integration of HBx in front of LINE1 element and the existence of HBx-LINE1 chimeric transcript in Hepatitis B virus-related HCC. All results in this article were analyzed with the software SPSS version 19.0 for Windows, and different groups were compared by χ2 test as appropriate.RESULTS: Based on the data from COSMIC database, compared with other solid tumors, mutation frequency of CTNNB1 was significantly higher in HCC(P < 0.01). The rate of CTNNB1 mutation was significantly less frequent in Hepatitis B virus-related HCC than in other etiologies(P < 0.01). Dual-luciferase reporter system and TOP/FOP reporter assays confirmed that TP53 GOF mutants were able to enhance the transcriptional ability of Wnt signaling. An exclusive relationship between the status of TP53 and CTNNB1 mutations was observed. However, according to the COSMIC database, TP53 GOF mutation is rare in HCC, which indicates that TP53 GOF mutation is not a reason for the aberrant activation of Wnt signaling in HCC. APC and AXIN1 were mutated in HCC. By using methylation sensitive restriction enzyme-quantitative PCR, hypermethylation of APC was
文摘目的:检测早期乳腺癌患者循环肿瘤细胞(circulating tumor cells,CTC)计数及其TP53、磷脂酰肌醇-3激酶(PI3K)基因突变情况,探讨基因突变结果与患者临床病理特征的相关性。方法:收集30例乳腺癌和良性乳腺肿瘤确诊患者的临床资料及血液、尿液标本,采用CellSearch法检测患者外周血CTC数目;采用微滴式数字PCR(droplet digital PCR,ddPCR)技术检测患者血、CTC和尿标本中TP53(p.R248Q、p.R273H、p.R175H)和PI3K(p.E545K、p.H1047R)基因位点突变情况;分析基因突变情况与患者临床病理特征(发病年龄、肿瘤大小、组织学分级、分子分型、腋窝淋巴结转移和Ki-67指数等)间的相关性。结果:最终纳入29例患者,患者血、CTC和尿标本中共获得435个基因检测结果,其中发生基因突变的82个(18.9%)。5种基因位点的突变率不同,差异有统计学意义(P<0.001)。突变率最高的基因是TP53(p.R175H),有29个突变(6.7%);突变率最低的是PI3K(p.E545K),仅发生1个(0.2%)突变。CTC、血和尿标本间5种基因位点的突变率差异有统计学意义(P=0.007,P<0.001,P=0.002),3类样本中突变率最高的基因都是TP53(p.R175H)。乳腺癌分子分型和Ki-67指数与CTC标本中TP53(p.R248Q)基因突变相关(P=0.003,P=0.028),而腋窝淋巴结转移可能与CTC标本中TP53(p.R175H)基因突变有关(P=0.058)。结论:早期乳腺癌患者的肿瘤分子分型和Ki-67指数与CTC中TP53(p.R248Q)基因突变相关,而腋窝淋巴结转移可能与CTC中TP53(p.R175H)基因突变相关,提示CTC标本中TP53基因突变是早期乳腺癌治疗方案选择及预后判断的潜在指标。
文摘基于整合药理学和实验验证探讨健脾除痰解毒方(Jianpi Chutan Jiedu Decoction,JCJD)干预肺癌的作用机制。采用中医药整合药理学研究平台(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine,TCMIP)V2.0,检索并获取健脾除痰解毒方靶标及功能信息,以及肺癌疾病靶标及功能信息,借助中医关联网络挖掘平台构建“疾病-方剂”关联网络,并通过网络拓扑特征计算、自定义多维关联网络可视化(方剂-中药材-核心靶标-通路-疾病)以及基因本体(GO)功能和依托Reactome数据库的Pathway信息进行富集分析,发现健脾除痰解毒方干预肺癌的关键网络靶标及其作用机制。最后通过动物实验进一步验证整合药理学结果。结果共获得健脾除痰解毒方干预肺癌的核心靶标53个;GO富集分析得到生物过程有20种条目,细胞组分有11种条目,分子功能有20种条目;Reactome Pathway富集分析得到11个条目,主要涉及呼吸电子传递、TP53调节代谢基因、细胞内受体的SUMO化、白细胞介素(IL)-4和IL-13通路等信号通路。动物实验结果显示与模型组比较,联合组肿瘤组织中乳酸含量明显下降(P<0.05),联合组可明显上调肿瘤组织中磷酸酯酶与张力蛋白同源物(PTEN)蛋白平均光密度值(P<0.05),降低丙酮酸激酶M2(PKM2)和己糖激酶2(HK2)蛋白平均光密度值(P<0.05);与模型组比较,联合组可明显上调肿瘤组织中肿瘤抑制蛋白(P53)、PTEN mRNA及蛋白表达(P<0.01,P<0.05),降低肿瘤组织中TP53诱导的糖酵解与凋亡调控因子(TIGAR)、葡萄糖转运蛋白1(GLUT1)、PKM2、HK2 mRNA及蛋白表达(P<0.01)。综上,健脾除痰解毒方通过TP53调节代谢基因来下调葡萄糖代谢,从而抑制肺癌增殖。
基金Supported by Foundation of Xiamen Science and Technology Bureau,Fujian Province,China,No.3502Z20074023
文摘AIM:To rapidly detect molecular alterations in different malignancies and investigate the possible role of Tp53,C-myc,and CCND1 genes in development of tumors in human organs and their adjacent normal tissues,as well as the possible relation between well-and poorly-differentiated tumors.METHODS:A tissue array consisting of seven different tumors was generated.The tissue array included 120 points of esophagus,120 points of stomach,80 points of rectum,60 points of thyroid gland,100 points of mammary gland,80 points of liver,and 80 points of colon.Expressions of Tp53,C-myc,and CCND1 were determined by RNA in situ hybridization.3' terminal digoxin-labeled anti-sense single stranded oligonucleotide and locked nucleic acid modifying probe were used.RESULTS:The expression level of Tp53 gene was higher in six different carcinoma tissue samples than in paracancerous tissue samples with the exception in colon carcinoma tissue samples(P < 0.05).The expression level of CCND1 gene was significantly different in different carcinoma tissue samples with the exception in esophagus and colon carcinoma tissue samples.The expression level of C-myc gene was different in esophagus carcinoma tissue samples(c2 = 18.495,P = 0.000),stomach carcinoma tissue samples(c2 = 23.750,P = 0.000),and thyroid gland tissue samples(c2 = 10.999,P = 0.004).The intensity of signals was also different in different carcinoma tissue samples and paracancerous tissue samples.CONCLUSION:Over-expression of the Tp53,CCND1,and C-myc genes appears to play a role in development of human cancer by regulating the expression of mRNA.Tp53,CCND1 and C-myc genes are significantly correlated with the development of different carcinomas.
文摘Tp53, a stress response gene, is involved in diverse cell death pathways and its activation is implicated in the pathogenesis of Parkinson＇s disease. However, whether the neuronal Tp53 protein plays a direct role in regulating dopaminergic （DA） neuronal cell death or neuronal terminal damage in different neurotoxicant models is unknown. In our recent studies, in contrast to the global inhibition of Tp53 function by phar- macological inhibitors and in traditional Tp53 knock-out mice, we examined the effects of DA-specific Tp53 gene deletion after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and methamphetamine exposure. Our data suggests that the Tp53 gene might be involved in both neuronal apoptosis and neuronal termi- nal damage caused by different neurotoxicants. Additional results from other studies also suggest that as a master regulator of many pathways that regulate apoptosis and synaptic terminal damage, it is possible that Tp53 may function as a signaling hub to integrate different signaling pathways to mediate distinctive target pathways. Tp53 protein as a signaling hub might be able to evaluate the microenvironment of neurons, assess the forms and severities of injury incurred, and determine whether apoptotic cell death or neuro- nal terminal degeneration occurs. Identification of the precise mechanisms activated in distinct neuronal damage caused by different forms and severities of injuries might allow for development of specific Tp53 inhibitors or ways to modulate distinct downstream target pathways involved.
