Novel Mutations of PRSS1 Gene in the Patients with Pancreatic Cancer among Han Population

Purpose: The aim of this study was to elaborate some novel mutations of PRSS1 gene in the patients with pancreatic cancer. Patients and Methods: There were 156 patients with pancreatic cancer and 220 unrelated individuals were studied as controls. The mutations of PRSS1 gene were analyzed by direct sequencing. K-ras Mutation Detection Kit was used to find the general k-ras gene disorder in the pancreatic cancer tissue. Then collected and analyzed the clinical data at the same time. Results: There were two patients carried novel mutations which is IVS 3 +157 G>C of PRSS1 gene from peripheral blood specimens and pancreatic cancer tissue. What’s more, we were surprised to find a novel complicated mutation of exon 3 in PRSS1 gene (c.409 A>G and c.416 C>T) in another young patient. The c complicated mutation made No.135 and No.137 amino acid transfer from Thr to Ala and Thr to Met respectively. All of the mutations weren’t found in the normal controls and no mutations of k-ras gene detected in the three patients. Conclusion: These observations imdicate that mutations of PRSS1 gene may be an important factor of pancreatic cancer.     

Novel mutation and polymorphism of PRSS1 gene in the Chinese patients with hereditary pancreatitis and chronic pancreatitis

Background Mutations in the cationic trypsinogen gene （PRSS1） have been detected in patients with hereditary pancreatitis （HP）. This study investigated the prevalence of the R122H （c.365G〉A）, A121T （c.361 G〉A） and D162D （c.488 C〉T） mutations or polymorphisms in the common, non-hereditary forms of chronic pancreatitis and in an HP family.
Methods DNA was prepared from blood samples of 54 patients with chronic pancreatitis （35 alcoholic, 17 idiopathic and 2 hereditary） and 120 normal controls. The PRSS1 genes were amplified by polymerase chain reaction （PCR） and their products were analyzed by sequencing and related clinical data were also collected.
Results A new polymorphism （c.488 C〉T） of PRSS1 was found in 25 patients with chronic pancreatitis （including one affected member of the HP family） and six members of the normal controls. The C/T genotype was significantly increased in chronic pancreatitis （OR： 16.379, 95% CI： 5.7522-52.3663）, the frequency of c.488 C〉T change was in according with the Hardy-Weinberg equilibrium, but it doesn＇t affect the clinical phenotype. The commonly reported change of R122H （c.365G〉A） was not detected in any of the study subjects, c.361 G〉A was found in 2 affected members and one unaffected carrier in an HP family. One of the affected members of an HP family had c.361 G〉A mutation and polymorphism （c.488 C〉T） in the PRSS1 gene at the same time. The patient＇s clinical values （C3, C4, CA19-9 and HbA1c） were higher than those of the other patients with chronic pancreatitis. The two patients with HP developed diabetes mellitus and their father died with pancreatic cancer.
Conclusion A new polymorphism （c.488 C〉T） in the PRSS1 gene is associated with chronic pancreatitis, but it did not affect the clinical phenotype while the A121T （c.361 G〉A） mutation in the gene shows a significant correlation in the patients with HP.     

新疆地区哈萨克族乳腺癌患者BRCA1和BRCA2基因的突变分析

目的：探讨新疆地区哈萨克族散发性乳腺癌中 BRCA1和 BRCA2基因的突变频率及分布情况。方法选择2005年1月-2013年10月新疆医科大学第一附属医院和附属肿瘤医院收治的来自新疆地区的哈萨克族乳腺癌患者86例（病例组）和哈萨克族健康女性70例（对照组）,由外周静脉血提取基因组 DNA,对乳腺癌易感基因1（Breast Carcinoma 1,BRCA1）外显子2、10、18、20和乳腺癌易感基因2（Breast Carcinoma 2,BRCA2）外显子10、11及外显子-内含子拼接区进行DNA直接测序,并鉴定突变位点。结果病例组和对照组共检测到的18个突变位点中,rs80357374、rs4987117为未知功能变异,其余均为多态性改变。在病例组和对照组中分别检测到195个和143个变异序列。病例组 rs1799950、rs1801499、rs1799944的分布频率明显高于对照组,差异有统计学意义（P ＜0．05）。在中国人群中尚未检测到 rs80357374,Intron18-268为新发突变位点,但在对照组中均检测到2个基因突变。两组中携带4个以上突变的患者人数占各组携带者的56．9％、39．6％,差异无统计学意义（P ＞0．05）。BRCA2突变频率高于BRCA1,但2个基因突变频率差异无统计学意义（P ＞0．05）。结论哈萨克族乳腺癌患者BRCA1和BRCA2突变频率高,且相当一部分受试对象同时携带多个突变位点,这可能是哈萨克族人群的特有遗传特征。rs1799950、rs1801499、rs1799944这3个位点可能增加哈萨克族人群罹患乳腺癌的风险,值得今后进一步研究验证。     

广东省家族性和早发性乳腺癌BRCA1基因突变的相关研究

目的 研究广东省家族性和早发性乳腺癌患者的BRCAI基因突变情况及其与ER、PR、HER2和ALN等表达的关系.方法 抽取广东省58例家族性和早发性乳腺癌患者的外周静脉血,提取基因组DNA,应用PCR技术对BRCA1基因的全部编码序列进行扩增,突变分析由变性高效液相色谱分析(DHPLC)进行预筛后,进行DNA测序方法证实.免疫组化法检测患者中ER、PR、HER2和ALN的表达情况,并分析其表达与BRCA1基因突变的关系.结果 58例乳腺癌患者中,2例年龄<35岁的患者发生BRCA1致病突变,而且其中1个为新发现的拼接点突变(331G→A).BRCA1基因突变位点与ER、PR、HER2和ALN的表达无关.结论 广东省早发性乳腺癌患者和家族性乳腺癌患者的BRCA1基因突变的发生率明显低于西方围家;331G→A致病突变位点可能是广东省早发性乳腺癌的特有突变位点;基因突变位点可能与ER、PR、HER2和ALN等组织学表达无关.     

散发性乳腺癌患者BRCA1基因突变检测

目的:研究散发性乳腺癌患者中,乳腺癌易感基因-1(BRCA1)的突变情况,并探讨其临床意义。方法:采用聚合酶链反应-单链构像多态性分析(PCR-SSCP)、标记染色双脱氧末端法DNA测序,对27例散发性乳腺癌患者以及正常对照组8例进行BRCA1基因全序列外显子突变检测。结果:27例患者中发现1例第16外显子基因突变,突变率为3.7%(1/27例);突变形式为4804C→G,突变结果引起单个氨基酸改变,使编码子1562由精氨酸代替脯氨酸。结论:在散发性乳腺癌患者中BRCA1基因突变率较低(3.7%),BRCA1可能通过突变以外的调节方式起作用。     

Spectrum of NSD1 gene mutations in southern Chinese patients with Sotos syndrome

Sotossyndrome(MIM117550),alsoknownascerebralgigantism,wasfirstdescribedin1964.1Sincethen,hundredsofcaseshavebeenreported.2,3It isadiseasewiththecombinationofsomaticovergrowth, advancedboneage,characteristicfacialappearanceand developmentaldelay.2 OvergrowthinSotossyndromeisusuallyevidentsince birth.Allthegrowthparametersareincreased,withthe headsizeremaininglargewhileheightandweightshowing atendencytonormalizewithadvancingage.The characteristicfacialfeaturesarehighprominentforehead, hypertel…     

人胰腺腺癌中抗癌基因P53的突变

用ＰＣＲ－ＳＳＣＰ分析及ＤＮＡ直接测序法检测了１６例胰腺腺癌的Ｐ５３基因５－８外显子，结果发现３７．５％有Ｐ５３基因点突变，突变分布于外显子５－８，无特殊突变倾向点。６例有Ｐ５３突变者，５例为１个等位基因突变，另１个等位基因丢失；１例同时含有突变型和隆型等位基因。     

胰腺癌基因改变的研究进展

大量研究表明人类癌症是一种多基因病,与胰腺癌发生有关的基因改变主要包括K-ras、c-myc、c-fos、c-erbB-2等癌基因;p53、p16、DPC4/SMAD4、DCC等抑癌基因;EGF、FGF、HGF、PDGF、VEGF、TGF-β等生长因子及其受体.常见为K-ras、p53、p16、DPC基因改变,但胰腺癌的发生是一多步骤的现象,基因改变的积累至关重要.     

中国南部人群家族性及早发性乳腺癌患者p53基因胚系突变分析

目的：分析p53基因在中国南方部分地区家族性和早发性乳腺癌中的突变位点及特征。方法：以中国南 方地区150例家族性和早发性乳腺癌患者为研究对象,提取静脉血基因组DNA,对p53基因的全部编码序列及外显子 与内含子拼接区进行扩增。 采用变性高效液相色谱进行预筛后,应用DNA测序分析和证实基因突变的结果。结果： 150例患者中,9例的p53编码区域共发现6种不同的p53变异,其中869_888ins20(插入突变)为新发现的致病性突变, 643_660del18(缺失突变)为已报道的致病性突变,91G>A,215C>G,537T>G和743G>A为已报道有致病意义的错义突 变位点。此外,还发现了第4外显子区域的同义突变141G>A及第3内含子区域的缺失突变IVS3+54_70del16和9个基因多 态性位点。家族性及早发性乳腺癌的p53总突变率为6.00%,其中家族性乳腺癌的p53突变率约为6.81%。早发性乳腺癌 突变率约为6.25%。结论：中国南部人群家族性乳腺癌患者的p53基因胚系总突变率高于国内外文献报道,首次发现 的插入突变869_888ins20的致病意义有待今后的功能学验证。缺失突变643_660del18丰富了国人p53基因突变数据库, 有可能是中国乳腺癌人群的特有突变。     

胰腺癌患者血清CEACAM1的测定及其诊断价值

目的研究肿瘤标志物CEACAM1对胰腺癌诊断价值。方法收集南方医院、珠江医院和佛山市第一人民医院2008年1月~2009年10月住院胰腺癌病人50例和慢性胰腺炎对照50例;采用ELISA测定各组患者血清CEACAM1水平;根据ROC曲线中敏感性和特异性之和最高点为参考,制定CEACAM1的截断值并计算ROC曲线下面积;根据肿瘤标志物诊断效率评估表评价各个肿瘤标志物对胰腺癌诊断效率。结果在胰腺癌病人中,CEACAM1的血清值和阳性率与慢性胰腺炎组比较有显著升高(P<0.05);根据制定的ROC曲线,CEACAM1在本次实验中血清中含量测定的截断值为13.835 ng/ml,曲线下面积为0.780(P<0.05)。胰腺癌敏感性CEACAM1>CA242>CA19-9(P>0.05);特异性CA242>CA19-9>CEACAM1(P<0.05)。结论 CEACAM1对诊断胰腺癌的敏感性高于CA19-9和CA242,但是其特异性较低,不能单独用于胰腺癌的诊断。     

一条全长胰腺癌相关新基因的初步研究

目的 对应用cDNA芯片技术筛选出的一条全长胰腺癌相关新基因进行鉴定.方法 对应用cDNA芯片技术筛选出的一条全长胰腺癌相关新基因进行测序和生物信息学分析,应用RT-PCR和Northern blot检测该基因在12例胰腺癌和癌旁正常胰腺组织以及4种胰腺癌细胞株中的表达情况.结果 生物信息学分析显示新基因定为于染色体4p15,有一包含501 bp的ORF,拟编码由166个氨基酸组成的蛋白质,其理论分子量为18 293.23,等电点为8.57.BLASTp分析发现,该蛋白与一种人类假想蛋白FLJ90013同源性大于90%,可能为该假想蛋白家族新成员.RT-PCR显示新基因在12例胰腺癌组织和4种胰腺癌细胞株中均表达,而在正常胰腺组织中未见表达.Northern blot分析得到相同的结果.结论 新基因与胰腺癌的发生、发展密切相关,可作为胰腺癌诊断和基因治疗的分子靶标.     

SF3B1基因在胰腺癌组织中的表达及其与肿瘤分期分化程度的相关性

目的 探讨剪切因子3B亚基1(SF3 B1)基因在胰腺癌组织中的表达及其与肿瘤分期、病理程度相关性.方法 回顾性分析2017年9月至2019年8月空军军医大学第一附属医院行根治术治疗的48例胰腺癌患者临床资料,对术中切除的癌组织及癌旁组织行免疫组化染色,比较SF3B1基因在胰腺癌、癌旁组织中的阳性率;根据TNM分期将胰...     

Detecting K- ras and p53 gene mutation from stool and pancreatic juice for diagnosis of early pancreatic cancer

目的：通过检测胰液、粪便癌基因K-ras和抑癌基因p53的突变,探索早期诊断胰腺癌的手段。方法：1994年至2000年5月住院和门诊患共201例,正常对照60例,通过聚合酶链反应-限制片段长度多态性（PCR-RFLP）方法检测胰液、粪便K-ras突变,PCR-SSCP方法检测p53突变。结果：胰腺癌患胰液K-ras突变率为87.8％（36/41）,胰腺良性病变为23.5％（4/17）。粪便K-ras扩增成功率为90.0％（235/261）。胰腺癌患粪便K-ras突变率为88.0％（66/75）,胰腺良性病变为51.1％（24/47）,正常人粪便K-ras突变率为19.6％（9/46）。胰腺癌胰液细胞p53突变率47.4％（18/38）,胰腺良性疾病为12.5％（2/16）。胰腺癌粪便p53突变率为37.1％（23/62）,慢性胰腺炎为19.1％（4/21）。结论：胰腺癌患胰液K-ras突变的敏感性、特异性高,可以作为胰腺癌诊断的辅助手段。粪便K-ras结合p53检测可以提高敏感性,在胰腺癌筛选中有潜在的价值,联合血清CA19-9等肿瘤标记物检测有助于提高胰腺癌早期诊断率。     

人胰腺癌组织中R—ras的表达及其临床意义

目的检测人类胰腺癌组织中R-ras的表达情况并探讨其临床意义。方法用免疫组织化学方法检测65例胰腺癌组织及癌旁组织中R-ras、P21ras和增殖细胞核抗原(PCNA)的表达,并对检测结果作统计学分析。结果胰腺癌组织中R-ras、P21ras和PCNA的阳性表达率均显著高于癌旁组织中的表达率(均P<0.01);R-ras阳性表达率与胰腺癌组织的病理分化程度无关(P>0.05);R-ras阳性表达率与P21ras和PCNA的阳性表达率均相一致(均P<0.01)。结论R-ras在原发性胰腺癌组织中表达增加,可能参与了胰腺癌的发病过程。     

外周血ctDNA基因突变对早中期非小细胞肺癌患者的诊断效能分析

目的 分析外周血循环肿瘤DNA(circulating tumor DNA, ctDNA)基因突变对非小细胞肺癌患者的诊断效能。方法 纳入2016年12月至2017年12月在河北大学附属医院初诊初治且行单孔胸腔镜手术治疗的50例非小细胞肺癌(non-small cell lung cancer, NSCLC)患者为研究对象。收集所有研究对象外周血后分离血浆,并提取ctDNA。 Illumina HiSeq 3000平台进行目标区域的高通量测序,并以基因突变频率作为ctDNA的检测分析指标。结果 50例患者ctDNA检测非小细胞肺癌的敏感性明显高于血清中癌胚抗原检测的敏感性(P<0.05);ctDNA基因突变频率在不同年龄、性别、病理类型、吸烟史等患者间差异无统计学意义(P>0.05),而在不同肿瘤大小及临床分期患者中差异有统计学意义(P<0.05)。结论 外周血ctDNA基因突变可敏感诊断早中期NSCLC,且与早中期NSCLC肿瘤大小及临床分期有相关性。     

基因表达谱芯片在胰腺癌相关基因筛选中的应用研究

目的：探讨基因表达谱芯片技术在高通量筛查肿瘤相关基因群及研究胰腺癌分子病理变化中的应用价值。方法：应用含有４０９６条人类全长基因的ｃＮＤＡ表达谱芯片,对３例临床切除的胰腺癌及正常胰腺标本的基因表达谱进行分析。结果：在３例胰腺癌和正常胰腺组织中均有差异表达的基因３９８条,其中新基因２８９条,老基因１０９条。从老基因中筛选出有显著表达差异基因３７条,其中在胰腺癌组织中上调基因２０条,下调基因１７条。结     

新疆维吾尔族非综合征型先天缺牙家系遗传分析及MSX1基因突变检测

目的探讨新疆维吾尔族非综合征型先天缺牙发病的分子机制。方法对2个维吾尔族先天缺牙家系绘制系谱图,分析家系遗传特征,并采集家系成员颊黏膜拭子,提取DNA,采用聚合酶链反应(PCR)技术结合DNA双向测序技术检测MSX1基因突变。结果 MSX1基因外显子1的353位点和外显子2的448位点检测出2个单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点。结论 MSX1基因外显子1的353位点的改变可能与新疆维吾尔族非综合征型先天缺牙的发生有关。     

Unique GGT→GTT mutation at K-ras codon 12 in six human pancreatic cancer cell lines from Chinese patients

Objective To investigate the K-ras mutation pattern in six pancreatic cancer cell lines from Chinese patients. Methods All six cell lines were analyzed for mutations in exon 1 of the K-ras gene by polymerase chain reaction (PCR) and direct sequencing.Results All 6 pancreatic cancer cell lines had GGT→GTT mutations at K-ras codon 12 but no mutations at codon 13.Conclusion The unique GGT→GTT mutation at codon 12 plays a potential role in the carcinogenesis of pancreatic cancers in Chinese.     

Adenoviral mediated suicide gene transfer in the treatment of pancreatic cancer

Objective To determine the efficacy of adenovirus- mediated suicide gene transduction comb ined with prodrug 5- fluorocytosine (5FC) as a therapeutic protocol for pancreat ic cancer. Methods Cytosine Deaminase(CD) gene was cloned into pAdTrack- CMV- CD, pAdTrack- CMV- CD and pAdEasy- 1 were recombined in bacteria. The newly recombined adenovirus (A d)- CD containing green fluorescent protein (GFP) were packaged and propagated i n 293 cells and purified by cesium chloride gradient centrifugation. Human panc reatic carcinoma cell line- Patu8988 was infected with this virus, then 5FC was added. XTT assay was used to estimate relative numbers of viable cells. In viv o model of pancreatic cancer was established by injecting 1. 0×10[7] Patu8988 c ells subcutaneously in Balb/c nude mice. When tumors were palpable, Ad- CD was injected into each tumor and 5FC was administered. Results Positive clones were selected using endonuclease to digest the recombinants and the concentration of viral liquids containing the CD gene was 2×10[11] pfu /ml. Significant cytotoxic activity as shown for 5FC in the CD gene transduced 8988 cell line, while little effect was found in the nontransduced pancreatic ca rcinoma cells. Antitumor effect was observed in Patu8988 xenograft nude mice wi th in situ CD gene transduction. Conclusions CD gene mediated by adenovirus has high infectivity and may be useful for gene t herapy in pancreatic carcinoma. These data demonstrate the use of an enzyme pro drug strategy in experimental pancreatic cancer.     

p53基因在肺癌患者中的突变

目的　探讨肺癌组织中p5 3基因蛋白表达和肺癌患者血白细胞p5 3基因突变及其相互关系。方法　应用免疫组织化学S -P法检测肺癌组织中p5 3基因蛋白表达及聚合酶链反应 -单链构象多态性分析 (PCR -SSCP)检测其中肺癌患者血白细胞p5 3基因突变。结果　p5 3基因蛋白在 5 1例肺癌组织中的阳性表达率为 5 9% ,其中p5 3蛋白在鳞癌、腺鳞癌和腺癌的表达差异无显著性 (P >0 .0 5 ) ,但有淋巴结转移者 ( 77% )显著高于无淋巴结转移者 ( 4 5 % ,P <0 0 5 ) ,p5 3蛋白在肺癌癌旁正常肺组织及白细胞中均为阴性 ;5 1例肺癌患者中 2 8例血白细胞DNA均未见p5 3基因突变。结论　p5 3基因在肺癌患者血白细胞DNA未见突变 ,p5 3基因蛋白阳性的肺癌患者更易发生淋巴结转移