Construction and Expression of Human PTEN Tumor Suppressor Gene Recombinant Adenovirus Vector

Phosphatase and tensin homologue deleted onchromosome10(PTEN)is a newtumor suppres-sor gene and possesses mutation in multiple kindsof tumors which include breast cancer[1].Teng[2]found the incidence for loss of heterozygosity(LOH)in breast cancer speci mens was48%(32/67),and the mutation and inactivation of PTENgene plays a central role in cell migration,growthandinvasion of breast cancer[3].This study ai ms atestablishing a kind of proliferative defective recom-binant adenovirus vector A…     

Transfection of Antisense Oligodeoxynucleotide Inhibits Heparanase Gene Expression and Invasive Ability of Human Pancreatic Cancer Cell in vitro

Cancer cell invasion and metastasis are criticalsteps in pancreatic cancer progression that worsenthe prognosis.They are facilitated via disassemblyof the main structural elements of basement mem-branes(BM)and extracellular matrix(ECM)byenzymes produced by the cancer cells[1].Hepara-nase(HPSE)is an endoglycosidase responsible forthe degradation of the heparan sulfate proteoglycan(HSPG),a major component of ECM and BM.HPSE mediates tumor invasion pri marily throughthe direct cleavage of…     

Genetic study of a large Chinese kindred with von Hippel-Lindau disease

Background Von HippeI-Lindau (VHL) disease is a heraditary cancer syndrome caused by germline mutations of the VHL tumor on the suppressor gene. This study was to show the clinical characteristics of a large Chinese kindred with yon HippeI-Lindau disease and to evaluate the role of the genetic test of VHL disease in the diagnosis of VHL disease and clinical screening of members of the VHL disease family.Methods DNA extracted from peripheral blood was amplified by PCR to three exons of the VHL gene in 27 members of a large kindred with VHL disease. PCR products were directly sequenced. The involvements of multi-organs in the kindred with VHL disease were confirmed by history taking and radiography.Results Of 47 members in the four generations of the kindred, 18 members were diagnosed as having VHL desease. Clinical manifestations of 18 patients included: central nervous system (CNS)hemangioblastoma (5), renal cell carcinoma and CNS hemangioblastoma (3), renal cell carcinoma and retinal angioma (3), renal cell carcinoma and multiple pancreatic cysts (1), renal cell carcinoma and retinal angioma and multiple pancreatic cysts (2), renal cell carcinoma and CNS hemangioblastomas and multiple pancreatic cysts (1), and multiple pancreatic cysts and multiple renal cysts (1), multiple pancreatic cysts (2). The common lesions of the 18 patients were renal cell carcinoma (55.6%), CNS hemangioblastoma (50.0%), retinal angioma (27.8%), and multiple pancreatic cysts (38.9%). Among the 27 members who volunteered for genetic analysis, 15 members including 9 affected family patients and 2 asymptomatic patients and 4 carriers, who are still alive, presented a codon 78 from Asn to Ser change at nucleotide 446 (A→G) in exon 1. Four members were carriers with the same VHL gene mutation. Two asymptomatic patients were initially diagnosed by genetic testing and subsequently confirmed radiologically and surgically. Members without gene mutation had no clinical evidence of VHL disease.Conclusions The large Chinese kindred with VHL disease was classified as type I . The main characteristics in the kindred were higher incidence of renal cell carcinoma and lower incidence of retinal angioma. Genetic test plays an important role in early detecting asymptomatic patients and the carriers in clinical screening of members of the families with VHL disease. It is also important to prevent the transmission of VHL disease to their offsprings in the kindred.     

REDUCED TUMORIGENICITY OF METASTATIC HUMAN LUNG CANCER CELL SUBLINE （PGCL3） TRANSFECTED WITH hRARβ GENE

The recombinant PSG5-RARβ plasmid and the G418 resistant PSVzneo plasmid (10:1) were cotransfected into PGCL3 cells by coprecipitation with calcium phosphate. The transfectants CR3 and CR4, which xpressed the RARβ gene, were identified by Northern blot hybridization. The results showed that the in vitro growth and invasion of CR3 and CR4 were dramatically reduced compared to the control transfected cell (CSV1). Furtherraore, the colony-forming abilities in soft agar and the tumorigenicity in nude mice of CR3 and CR4 were abrogated, Our results suggests that RARβ functions not only as a receptor mediating the RA action, but also as a suppressor in lung tumorigenesis.     

Expression of p63 and Cyclooxygenase-2 and Their Correlation in Skin Tumors

To study the expression of p63 and cyclooxygenase-2 (cox-2) in skin tumors and evalu- ate the correlation between p63 and cox-2, the expressions of cox-2 and p63 were measured by streptavidin-peroxidase complex immunohistochemical technique in 17 cases of skin squamous cell carcinoma(SCC), 19 cases of Bowen's disease(Bowen), 11 cases of actinic keratosis(AK), 12 cases of seborreic keratosis(SK) and 13 specimens of normal skin. Our results showed that the expression of p63 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while the expression of p63 in seborreic keratosis was sig- nificantly higher than that in normal skin. The expression of cox-2 in skin squamous cell carcinoma, Bowen's disease and actinic keratosis were significantly higher than that in seborreic keratosis, while no statistical difference was noted in the expression of cox-2 between seborreic keratosis and normal skin. Cox-2 expression was positively correlated with the high p63 expression in malignant skin tu- mors. The increased expression of cox-2 and p63 may play an important role in the development of skin tumors and work synergetically in malignant skin tumors.     

Polymorphisms in tumor necrosis factor genes and susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in a population of high incidence region of North China

Background We investigated the possible association of the functional polymorphisms in the tumor necrosis factor (TNF) genes with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA). Methods The TNF-α-308G/A and TNF-β+252G/A single nucleotide polymorphisms (SNPs) were genotyped using polymerase-chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, in 555 cancer patients (291 ESCC and 264 GCA) and 437 healthy controls in a high incidence region of North China. Results Among healthy controls, frequencies of the TNF-α 1/1, 1/2 and 2/2 genotypes were 89.4%,9.2% and 1.4% respectively, while frequencies of the TNF-β B1/B1, B1/B2 and B2/B2 genotypes were 12.6%,32.3% and 55.1%, respectively. No significant difference was found in the overall genotype and allelotype distribution of the TNF-α-308G/A and TNF-β+252G/A SNPs among cancer patients and controls. However, both the B1/B1 genotype and B1/B2 genotype significantly increased the risk of developing ESCC ［the age and gender adjusted odds ratio (OR)=2.04 and 1.91, 95% confidence interval (CI)=1.04-4.43 and 1.14-2.60, respectively］ and GCA (the age and gender adjusted OR=2.68 and 2.64, 95% CI=1.14-6.29 and 1.47-4.72, respectively) in individuals with negative family history of UGIC, in comparison with the B2/B2 genotype. When the two TNF polymorphisms were combined and analyzed, individuals with the TNF-β B1/B2 and TNF-α1/2 or 2/2 genotypes significantly reduced the risk of developing ESCC and GCA, in comparison with those harboring the TNF-β B2/B2 and TNF-α 1/1 genotypes (the age and gender adjusted OR=0.37 and 0.34, 95% CI=0.15-0.92 and 0.13-0.90, respectively). ConclusionsTherefore, the TNF-α-308G/A and TNF-β+252G/A genotyping may be used as a stratification markers to predicate the risk of ESCC and GCA development in North China. Chin Med J 2005; 118(22):1870-1878     

Effects of Gene Tranfection with CH50 Polypeptide on the Invasion Ability of Bladder Cancer Cell Line BIU-87

Fibronectin(FN) ,a glycoprotein,possesses mul-tiple cell bioactivities such as influencing the cellularmorphology, controlling cellular migration,inducingcellular differentiation and affecting i mmune cell func-tions etc ,whichis closelyrelated withthe carcinogene-sis ,invasion and metastasis[1].CH50 polypeptide is composed of Cell I andHep II bifunctional-domain of human FN, main-tains the function of the original structure and doesnot formnovel function because of deletion of frag-ments a…     

ASSOCIATION BETWEEN LOW-DENSITY LIPOPROTEIN RECEPTOR-RELATED PROTEIN GENE, BUTYRYLCHOLINESTERASE GENE AND ALZHEIMER‘S DISEASE IN CHINESE

INTRODUCTION Alzheimer's disease (AD), an insidiously progressive dementia primarily affecting the elderly, is characterized clinically by gradual memory impairment and loss of pre- symptomatic persona leading ultimately to death. The neuropathological hallmarks of the disease are primarily senile plaques (SPs) and neurofibrillary tangles (NFTs). Genetic factors are of importance in the cause of AD. Mutations in the amyloid precursor protein (APP), presenilin- L(PS- 1) and presenili…     

MACC1基因在肝细胞癌中的表达及临床意义

目的 探讨MACC1基因在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及其临床意义.方法 应用逆转录聚合酶链反应(RT-PCR)方法 检测42例肝癌组织、对应癌旁肝组织以及17例正常肝组织中MACC1 mRNA的表达情况,并分析MACC1基因的表达与相关临床参数的关系.结果 MACC1基因在肝癌组织的表达水平显著高于癌旁肝组织(P＜0.01),在癌旁肝组织的表达水平显著高于正常肝组织(P＜0.05),且MACC1基因的表达水平与肝癌的TNM分期、肝内或淋巴结转移、门静脉癌栓等均明显相关(均P＜0.05),而与肿瘤个数、血清甲胎蛋白(AFP)水平、有无肝硬化等无明显关系(均P＞0.05).结论 MACC1基因可能在肝细胞癌的侵袭转移中发挥着重要作用,有可能成为肝癌治疗的新靶点之一.     

Glutathione S-transferase M1 and T1 genotypes and endometriosis risk: a case-controlled study

Objective To investigate the correlation between glutathione S-transferase (GST) M1 and T1 genotypes and endometriosis risk (EM). Methods Polymerase chain reaction (PCR) technique was used to detect the presence or absence of the GSTM1 and GSTT1 genes in genomic DNA isolated from the blood samples of 68 Han Chinese women with endometriosis and 28 without endometriosis. Results The frequencies of GSTM1 and GSTT1 null genotypes in women with endometriosis were 0.721 (49/68) and 0.779 (53/68), respectively, and in women without endometriosis were 0.429 (12/28) and 0.321 (9/28), respectively. There was a significant difference with regard to the frequencies of GSTM1 and GSTT1 null genotypes between the women with and without endometriosis (P<0.01). Furthermore, the frequencies of GSTM1 and GSTT1 null genotypes were significantly higher in the patients with stage Ⅲ and Ⅳ endometriosis ［0.731 (38/52) and 0.788 (41/52), respectively］ than in women without endometriosis (P<0.01), and the frequency of GSTT1 null genotype was statistically higher in patients with stage Ⅰ and Ⅱ endometriosis ［0.75 (12/16)］ than in the women without endometriosis (P<0.01). No correlation between GSTM1 and GSTT1 null genotypes and age, induced abortion or dysmenorrhea was detected in this study (P>0.05). Conclusion GSTM1 and GSTT1 null genotypes may be risk factors for the development of endometriosis.     

Relationship between the Expression of MTA-1 Gene and the Metastasis and Invasion in Human Osteosarcoma

Summary：To compare the expression level of metastasis associated-1 （MTA 1 ） gene in high and low metastatic： human osteosarcoma cell lines and examine the relationship of MTA 1 expression and the metastasis potentiality of osteosarcoma cells, the expression of MTA 1 in MC-63 osteosarcoma cell lines with high and low metastasis potential was detected by semiquantitative TR-PCR. Boyden chamber invasion assay was used to evaluate the invasive capacity in vitro in two osteosarcoma cell lines, The low metastasis MG-63 cells were transfected with MTA 1 full-length cDNA expression plasmid by lipofectamine and the changes of MTA 1 expression and in vitro invasion potential were examined after the transfection. Our results showed that MG63 cell line with high metastasis potential expressed significantly higher MTA 1 than that of MG63 cells with low metastasis as reavealed by RT-PCR The invasion potential of low metastasis MG63 cell line was increased after MTA 1 gene transfection. It is concluded that there may be a relationship between MTA 1 and invasive potentiality of human osteosarcoma cells, and the mechanism of MTA 1 in osteosarcoma metastasis and its possible role in associated gene therapy deserve further study.     

SUPPRESSION OF MALIGNANT PHENOTYPE OF A TRANSFORMED MOUSE MAMMARY EPITHELIAL CELL LINE （11A1） BY TUMOR SUPPRESSOR GENE RB

A malignant transformed mammary epithelial cell line (11 A1) was transfected with liposome encapsulated eukaryotic expression plasrrdd pCMV-neo-RB, yielding 4 constant clones which have obvious phenotypic reversion changes, and named 11Al-R1～R4 respectively. Further experiments showed that the llA1-R1 behaved like normal epithelial cells in both morphological and hiolcgical characteristics, with decreased clonogenicity in solid argar medium as well as decreased tumorigenicity. Northern blot hybridization showed increased expression of RB gene and decreased expression of c myc gene in llA1-R1, 11A1-R2 cells compared to llA1 cells. This was an ideal phenotypic reversion model for epithelial transformed cell line and demonstrated that the RB gene can reexpress and suppress malignant phetlotype in RB inactive cells.     

Interleukin- 1 gene polymorphism disease activity and bone mineral metabolism in rheumatoid arthritis

Objective To determine whether interleukin- 1α and 1β gene polymorphism is associated w ith rheumatoid arthritis disease activity and bone mineral metabolism, and whet her there is any relationship between IL- 1β and rheumatoid arthritis (RA) moti f gene. Methods IL- 1 gene polymorphisms were analyzed in 65 RA patients who met American College of Radiology (ACR) criteria and 60 controls. From genomic DNA, 2 polymo rphisms in each gene for IL1α- 889 and IL- 1β+3953 were typed by PCR- RFLP and HLA- DRB1 allele typing was also undertaken by PCR- SSOP. Some clinical and l aboratory parameters were collected. The allelic frequencies and carriage rate s were compared between RA patients and controls and between patients with acti ve and quiescent disease. Comparison was also made between IL- 1 polymorphism a nd parameters of bone mineral metabolism and between patients with the HLA- DRB1 RA motif plus IL- 1β2 and patients without the two alleles. Fisher test an d the analysis of variance was used to analyze the data. Results There was no significant difference in the frequency and carriage rate of IL- 1 α polymorphisms between RA patients and the controls. The β2/2 genotype of IL - 1β was more common in female RA patients compared with controls (P=0. 001 ). A lower carriage rate of IL- 1β2 occurred in male RA patients (P=0. 0 01). A higher carriage rate of IL- 1α2 is associated with a higher ESR (P =0. 008), HAQ score (P=0. 03), and vit- D(3) (P<0. 001), but conversely a lower SJC (p=0. 002), a lower RF (P=0. 002) and a lower BMD at the l um bar spine (P=0. 001). A higher frequency of IL- 1α1 is associated with a l ow er CRP value (P=0. 009). An increased IL- 1β2 carriage is associated with active rheumatoid disease as indicated by a higher CRP (P<0. 001), ESR ( P<0. 001) and pain score (P=0. 001) and a higher BMD at the lumbar spin e (P=0. 007), lower vit- D(3) and. Udpd/Crea level The presence of the HLA DR B1 RA motif and IL- 1β allele 2 at same time did not contribute to disease acti vity. Conclution Polymorphisms of the IL- β gene may affect the RA occurrence. Carriage of IL- 1β2 polymorphisms is associated with more active disease in RA and the presenc e of both the IL- 1α2 and the IL- 1β1 allele in RA influences bone resorption .     

p21WAF1/CIP1 gene DNA sequencing and its expression in human osteosarcoma

Background Mutation and expression change of p21^WAF1/CIP1 may play a role in the growth of osteosarcoma. This study was to investigate the expression of the p21^WAF1/CIP1 gene in human osteosarcoma, p21^WAF1/CIP1 gene DNA sequence change and their relationships with the phenotype and clinical prognosis.Methods p21^WAF1/CIP1 gene in 10 normal people and the tumours of 45 osteosarcoma patients were examined using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) with silver staining. The PCR product with an abnormal strand was sequenced directly. The p21^WAF1/CIP1 gene mRNA and P21 protein of 45 cases of osteosarcoma were investigated by using in situ hybridization and immunohistochemistrv, respectively.Results The occurrence of P21 protein in osteosarcoma was 17. 78% (8/45), and p21^WAF1/CIP1 mRNA expression in osteosarcoma was 42.22% (19/45). The p21^WAF1/CIP1 gene DNA sequencing of amplified production showed that in p21^WAF1/CIP1 gene exon 3 of 36 cases of human osteosarcoma,there were 17 cases (47.22%) with C→T at position 609; 10 normal blood samples‘ DNA sequence analysis yielded 8 cases (80.00%) with C→T at the same position.Conclusions Along with the increase of malignancy, the expression of p21^WAF1/CIP1 mRNA and P21 protein in osteosarcoma tends to decrease. It is uncommon for the p21^WAF1/CIP1 gene mutation to occur in human osteosarcoma. As a result, the possible existence of tumour subtypes of p21^WAF1/CIP1 gnemutation should be investigated. Our research leads to the location of p21^WAF1/CIP1 gene polymorphism of Chinese osteosarcoma patients, which can provide a basis for further research.     

nm23-H1、p16抑癌基因蛋白在甲状腺肿瘤中的表达及其临床意义

目的:探讨nm23-H1、p16抑癌基因蛋白在甲状腺肿瘤中的表达意义.方法应用SP免疫组化方法检测了54例甲状腺癌、16例腺瘤、10例甲状腺非瘤病变及癌旁正常组织中nm23-H1、p16抑癌基因蛋白的表达.结果 nm23-H1、p16基因在正常组织中阳性表达率为分别为100%(12/12)及91.7%(11/12),但两者均以弱表达为主,并且阳性着色均定位于胞核.甲状腺腺瘤及分化型甲状腺癌中nm23-H1、p16表达率及表达强度均增强(P<0.05),而甲状腺未分化癌中nm23-H1及p16表达率及表达强度则降低(P<0.01).nm23-H1及p16蛋白表达与甲状腺癌的分化程度呈正相关(P<0.01),与颈淋巴结转移无相关性,但nm23-H1与包膜侵犯呈负相关(P<0.05),nm23-H1、p16在甲状腺癌中的表达有相关关系(P<0.01).结论 nm23-H1及 p16在甲状腺癌的表达中有相关关系(P<0.01).以nm23-H1及p16蛋白失活与甲状腺未分化癌的发生密切相关,但与颈淋巴结转移无关.nm23-H1及p16可作为临床判断恶性程度及生物学行为的一个参考指标.     

Apolipoprotein E polymorphism in the early onset of coronary heart disease

Objective To assess the relationship between apolipoprotein E (apoE) polymorphism and the early onset of coronary heart disease (CHD) and the effect of apoE on lipids and lipoproteins in healthy Chinese subjects. Methods Sixty-eight patients with CHD younger than 55 years (CHD1), 136 patients with CHD older than 65 years (CHD2), and 136 healthy subjects were enrolled, and their plasma levels of triglyceride (TG), total cholesterol (TC) and high density lipoprotein cholesterol (HDL-C) were determined. The apoE genotypes were identified by polymerase chain reaction-restriction fragment length polymorphism.Results apoE 3/4 genotype and E4 allele frequency in the CHD1 group were higher than those in the CHD2 group and healthy subjects, while no differences were found between CHD2 and healthy subjects. Meanwhile, the plasma levels of TC and low density lipoprotein cholesterol (LDL-C) were higher in the CHD2 group than in both CHD1 group and healthy subjects.Each apoE isoprotein has variable TC and LDL-C levels that is E2 (E2/2+E2/3)＜E3(E3/3)&lt;E4(E4/4+E3/4). Conclusion apoE is one of the genetic factors that affect TC and LDL-C levels, and apoE 4 has a very close relation to CHD, suggesting that apoE 4 is an independent genetic factor of the early onset of CHD.     

转移性和非转移性食管癌组织中KAI1基因的表达

目的:探讨KAI1基因与食管癌转移的关系.方法:应用原位杂交法分析KAI1基因在有淋巴结转移和无淋巴结转移的食管癌组织中的表达.结果:在有淋巴结转移和无淋巴结转移的食管癌组织中KAI1 mRNA的表达未见显著差异 (P>0.05),且KAI1 mRNA表达与食管癌的分化无关(P>0.05).结论:KAI1基因对食管癌转移的发生无抑制作用;KAI1基因的表达也与食管癌的分化无关.