CLONING AND SEQUENCING OF IMMUNOGLOBULINVARIABLE－REGION GENE OF A MONOCLONALANTIBODY SPECIFIC FOR HUMANHEPATOCARCINOMA

ＣＬＯＮＩＮＧＡＮＤＳＥＱＵＥＮＣＩＮＧＯＦＩＭＭＵＮＯＧＬＯＢＵＬＩＮＶＡＲＩＡＢＬＥ－ＲＥＧＩＯＮＧＥＮＥＯＦＡＭＯＮＯＣＬＯＮＡＬＡＮＴＩＢＯＤＹＳＰＥＣＩＦＩＣＦＯＲＨＵＭＡＮＨＥＰＡＴＯＣＡＲＣＩＮＯＭＡＹａｎｇＰｉｎｇ杨萍；ＧａｏＬｅｉ高...     

THE EFFECT OF ACTIVE COMPONENTS OF LYCIUM BARBARUM AND GARLIC（LB－GO）ON THE SYNTHESIS OF DNA AND ULTRASTRUCTURE OF U_（14） CERV

ＴＨＥＥＦＦＥＣＴＯＦＡＣＴＩＶＥＣＯＭＰＯＮＥＮＴＳＯＦＬＹＣＩＵＭＢＡＲＢＡＲＵＭＡＮＤＧＡＲＬＩＣ（ＬＢ－ＧＯ）ＯＮＴＨＥＳＹＮＴＨＥＳＩＳＯＦＤＮＡＡＮＤＵＬＴＲＡＳＴＲＵＣＴＵＲＥＯＦＵ_（１４）ＣＥＲＶＩＸＣＡＮＣＥＲＣＥＬＬＳＩＮＭＩＣ?..     

PROMOTION OF CHEMICAL CARCINOGENESIS AND P53 EXPRESSION BY REDUCTION OF SUPEROXIDE DISMUTASE ACTIVITY IN THE LUNG OF RAT IN V

ＰＲＯＭＯＴＩＯＮＯＦＣＨＥＭＩＣＡＬＣＡＲＣＩＮＯＧＥＮＥＳＩＳＡＮＤＰ５３ＥＸＰＲＥＳＳＩＯＮＢＹＲＥＤＵＣＴＩＯＮＯＦＳＵＰＥＲＯＸＩＤＥＤＩＳＭＵＴＡＳＥＡＣＴＩＶＩＴＹＩＮＴＨＥＬＵＮＧＯＦＲＡＴＩＮＶＩＶＯ￥ＹｕＬｕｎｙｉｎ；喻伦银；Ｂｉ...     

ANTITUMOR EFFECT OF INTRATUMORAL INJECTION OF LIPOSOME-ENCAPSULATED G-CSF GENE AND IN SITU BIOLOGICAL CHARACTERISTICS OF THE TREATED TUMOR CELLS

ＡＮＴＩＴＵＭＯＲＥＦＦＥＣＴＯＦＩＮＴＲＡＴＵＭＯＲＡＬＩＮＪＥＣＴＩＯＮＯＦＬＩＰＯＳＯＭＥＥＮＣＡＰＳＵＬＡＴＥＤＧＣＳＦＧＥＮＥＡＮＤＩＮＳＩＴＵＢＩＯＬＯＧＩＣＡＬＣＨＡＲＡＣＴＥＲＩＳＴＩＣＳＯＦＴＨＥＴＲＥＡＴＥＤＴＵＭＯＲＣＥＬＬ...     

ENHANCED ANTITUMOR EFFECTS OF SUICIDE GENE THERAPY BY SIMULTANEOUS TRANSFER OF GMCSF GENE IN LEUKEMIA-BEARING MICE

ＥＮＨＡＮＣＥＤＡＮＴＩＴＵＭＯＲＥＦＦＥＣＴＳＯＦＳＵＩＣＩＤＥＧＥＮＥＴＨＥＲＡＰＹＢＹＳＩＭＵＬＴＡＮＥＯＵＳＴＲＡＮＳＦＥＲＯＦＧＭＣＳＦＧＥＮＥＩＮＬＥＵＫＥＭＩＡＢＥＡＲＩＮＧＭＩＣＥ１ＪｕＤｉａｎｗｅｎ鞠佃文ＣａｏＸｕｅｔａｏ２曹雪...     

THE THERAPEUTIC EFFECT OF INTRATUMORAL INJECTION OF GM-CSF GENE-MODIFIED ALLOGENIC MACROPHAGES ON TUMOR-BEARING MICE

ＴＨＥＴＨＥＲＡＰＥＵＴＩＣＥＦＦＥＣＴＯＦＩＮＴＲＡＴＵＭＯＲＡＬＩＮＪＥＣＴＩＯＮＯＦＧＭＣＳＦＧＥＮＥＭＯＤＩＦＩＥＤＡＬＬＯＧＥＮＩＣＭＡＣＲＯＰＨＡＧＥＳＯＮＴＵＭＯＲＢＥＡＲＩＮＧＭＩＣＥＹｕＹｉｚｈｉ于益芝ＣａｏＸｕｅｔａｏ?..     

A MICROCOMPUTER PROGRAM FOR CALCULATINGTHE CONFIDENCE INTERVALS OF SURVIVAL PROBABILITY IN MEDICAL FOLLOW－UP STUDIES

ＡＭＩＣＲＯＣＯＭＰＵＴＥＲＰＲＯＧＲＡＭＦＯＲＣＡＬＣＵＬＡＴＩＮＧＴＨＥＣＯＮＦＩＤＥＮＣＥＩＮＴＥＲＶＡＬＳＯＦＳＵＲＶＩＶＡＬＰＲＯＢＡＢＩＬＩＴＹＩＮＭＥＤＩＣＡＬＦＯＬＬＯＷ－ＵＰＳＴＵＤＩＥＳＸｉａｎｇｙｏｎｇｂｉｎｇ项永兵；Ｇａｏｙｕ...     

ESTABLISHMENT OF IMMUNO－PCR TECHNIQUE FOR THE DETECTION OF TUMOR ASSOCIATED ANTIGEN MG＿7－Ag ON THE GASTRIC CANCER

任军，樊代明，周绍娟，张学庸，杨安钢，李明峰，陈峥ＥＳＴＡＢＬＩＳＨＭＥＮＴＯＦＩＭＭＵＮＯ－ＰＣＲＴＥＣＨＮＩＱＵＥＦＯＲＴＨＥＤＥＴＥＣＴＩＯＮＯＦＴＵＭＯＲＡＳＳＯＣＩＡＴＥＤＡＮＴＩＧＥＮＭＧ＿７－ＡｇＯＮＴＨＥＧＡＳＴＲＩＣＣＡＮＣＥＲ￥Ｒ...     

STUDY ON THE PRECANCEROUS ESOPHAGEAL LESIONSOF RHESUS MONKEYS FROM TAIHANG AREAOF HIGH MORBIDITY OF ESOPHAGEAL

ＳＴＵＤＹＯＮＴＨＥＰＲＥＣＡＮＣＥＲＯＵＳＥＳＯＰＨＡＧＥＡＬＬＥＳＩＯＮＳＯＦＲＨＥＳＵＳＭＯＮＫＥＹＳＦＲＯＭＴＡＩＨＡＮＧＡＲＥＡＯＦＨＩＧＨＭＯＲＢＩＤＩＴＹＯＦＥＳＯＰＨＡＧＥＡＬＺｈａｎｇＨｏｎｇｘｕ张红绪；ＺｈａｏＸｉａｏｊｉｎ赵晓进...     

PROLIFERATION OF ANTI－CD_3 McAb AND IL－2 INDUCED SPLENOCYTES AND ANTITUMOR EFFECT OF THEIR CULTURE SUPERNATANTS

ＰＲＯＬＩＦＥＲＡＴＩＯＮＯＦＡＮＴＩ－ＣＤ_３ＭｃＡｂＡＮＤＩＬ－２ＩＮＤＵＣＥＤＳＰＬＥＮＯＣＹＴＥＳＡＮＤＡＮＴＩＴＵＭＯＲＥＦＦＥＣＴＯＦＴＨＥＩＲＣＵＬＴＵＲＥＳＵＰＥＲＮＡＴＡＮＴＳ￥ＳｈｅｎＧｕａｎｘｉｎ；沈关心；ＷａｎｇＸｉａｌｉｎ；?..     

反义核酸对肿瘤的治疗作用

鸟氮酸脱羧酶（ODC）是多胺生物合成途径中的限速酶．其活性的异常升高及多胺堆积与恶性肿瘤的发生、发展及转移密切相关。作者构建一个能表达人ODC反义RNA的真核表达质粒pCMV／ODCas，用磷酸钙沉淀法将其导入人肺鳞癌细胞系LTEP-78(L78）经G418筛选，获两株转染细胞系L78／ODCasl和L78／ODCas4。与亲本细胞相比，其生长速度减慢，软琼脂集落形成能力减弱，DNA、RNA及蛋白质等生物合成受到抑制，ODC酶活性降低。经Southern blot及PCR分析，进一步证明了上述变化与ODC反义序列稳定完整地整合到转染细胞基因组中有关。     

Single-injection ornithine decarboxylase-directed antisense therapy using atelocollagen to suppress human cancer growth

BACKGROUND: Substantial evidence supports a direct role of ornithine decarboxylase (ODC) in the development and maintenance of human tumors. Although antisense oligonucleotide therapy targeting various genes are useful for cancer treatment, 1 of the major limitations is the problem of delivery. A novel antisense oligonucleotide delivery method is described that allows prolonged sustainment and release of ODC antisense oligonucleotides in vivo using atelocollagen. METHODS: The effect of ODC antisense oligonucleotides in the atelocollagen on cell growth of gastrointestinal cancer (MKN 45 and COLO201) and rhabdomyosarcoma (RD) was studied in vitro using a cell-counting method with a hemocytometer. In vivo, the effect of intratumoral, intramuscular, and intraperitoneal single administration of ODC antisense oligonucleotides in the atelocollagen on tumor growth of MKN45, COLO201, and RD cells was studied. ODC activity and polyamine contents were measured. RESULTS: In vitro, ODC antisense oligonucleotides in the atelocollagen remarkably suppressed MKN45, COLO201, and RD cell growth. A single administration of antisense oligonucleotides in the atelocollagen via 3 routes remarkably suppressed the growth of MKN45, COLO201, and RD tumor over a period of 35-42 days. CONCLUSIONS: As various human cancers significantly express ODC, the results strongly suggest that this new antisense method may be of considerable value for treatment of human cancers.     

正反义人乙酰肝素酶真核表达体系构建及其意义的研究

目的:构建正反义人乙酰肝素酶(HPA)的肝癌细胞克隆表达体系,研究其对肝癌细胞转移潜能的影响。方法:建立正反义人HPA真核表达载体,脂质体法转染高转移人肝癌细胞系HCC-P,G418选择培养,以RT-PCR和免疫组织化学检测HPA mRNA及蛋白表达。细胞计数,MTT实验测定细胞生长状况。裸鼠尾静脉注射转染肿瘤细胞,30 d后称肺脏质量,计数肺表面转移结节数,比较不同转染细胞的转移潜能。结果:限制性内切酶及测序证实载体构建成功;RT-PCR证实,转染正反义载体的细胞在HPA mRNA水平上相差10.13倍;免疫组织化学证实,HPA高效稳定表达于转染正义载体的细胞胞质及胞膜上;细胞生长曲线和MTT实验表明,转染基因对细胞生长无明显影响,P>0.05。注射转染反义肝癌细胞组荷瘤鼠肺脏质量和肺表面转移结节数较对照组显著减少,正义组则相反,P<0.01。结论:成功克隆正反义人HPA,并稳定转染HCC-P。HPA基因对肝癌细胞转移潜能具有促进作用,而反义基因则抑制肿瘤的生长。     

Antizyme inhibitor 1: a potential carcinogenic molecule

Polyamines are multivalent and organic cations essential for cellular growth, proliferation, differentiation, and apoptosis. Increased levels of polyamines are closely associated with numerous forms of cancer. An autoregulatory circuit composed of ornithine decarboxylase (ODC), antizyme (AZ) and antizyme inhibitor (AZI) govern the intracellular level of polyamines. Antizyme binds with ODC to inhibit ODC activity and to promote the ubiquitin‐independent degradation of ODC. Antizyme inhibitor binds to AZ with a higher affinity than ODC. Consequently, ODC is released from the ODC–AZ complex to rescue its activity. Antizyme inhibitor increases the ODC activity to accelerate the formation of intracellular polyamines, triggering gastric and breast carcinogenesis as well as hepatocellular carcinoma and esophageal squamous cell carcinoma development. Antizyme inhibitor 1 (AZIN1), a primary member of the AZI family, has aroused more attention because of its contribution to cancer. Even though its conformation is changed by adenosine‐to‐inosine (A→I) RNA editing, it plays an important role in tumorigenesis through regulating intracellular polyamines. Encouragingly, AZIN1 has been revealed to have an additional function outside the polyamine pathway so as to bypass the deficiency of targeting the polyamine biosynthetic pathway, promising to become a critical target for cancer therapy. Here, we review the latest research advances into AZIN1 and its potential contribution to carcinogenesis.     

Specific inhibition of K-ras expression and tumorigenicity of lung cancer cells by antisense RNA

A human lung cancer cell line (H460a) with a homozygous spontaneous K-ras mutation was transfected with a recombinant plasmid that synthesizes a 2-kilobase genomic segment of the K-ras protooncogene in antisense orientation. Translation of the mutated K-ras mRNA in H460a cells was specifically inhibited, whereas expression of H-ras and N-ras was unchanged. A 3-fold growth inhibition occurred in H460a cells when expression of the mutated ras p21 protein was down-regulated by antisense RNA. However, cells remained viable despite the absence of K-ras expression. The growth of H460a tumors in nu/nu mice was substantially reduced by expressed K-ras antisense RNA.     

二氟甲基鸟氨酸及反义bcl-2协同诱导HL60细胞凋亡

目的　观察二氟甲基鸟氨酸 (DFMO)及反义bcl 2调控bcl 2基因表达对HL6 0细胞凋亡的诱导作用。方法　构建反义bcl 2逆转录病毒重组体 ,建立产病毒细胞系 ,转染HL6 0细胞 ,用形态观察、生长曲线、FCM分析、集落形成、DNA电泳图谱、分子杂交及免疫组化等方法研究细胞生长特性及细胞凋亡诱导。结果　成功地构建了反义bcl 2逆转录病毒重组体及产病毒细胞系。以此转染HL6 0细胞 ,引起了bcl 2mRNA及蛋白表达下降 ,但生长速率、细胞周期分布及鸟氨酸脱羧酶 (ODC)基因表达水平与亲本细胞比较差别不大 ;无明显的细胞凋亡 ,仅集落形成能力有所减弱。用小剂量DFMO处理反义bcl 2转染的细胞 ,不仅生长受到明显抑制 ,而且可诱导细胞凋亡。结论　DFMO与反义bcl 2对HL6 0细胞增殖抑制及凋亡诱导有协同作用。抑制bcl 2表达能提高肿瘤细胞对DFMO作用的敏感性。     

嵌合性启动子HRE.CArG的构建及对肝癌细胞乏氧和射线应答的调控

实体瘤中乏氧区的存在是影响放疗效果最主要的因素之一[1].在一个基因启动子内利用乏氧和射线应答元件联合,限制治疗基因仅在乏氧和/或受照组织激活,可提高治疗比率.该文研究目的是构建包含HREs和CArG元件的乏氧-辐射嵌合性基因启动子,并观察乏氧及照射时该启动子诱导增强的绿色荧光蛋白(enhanced green fluorescent protein,EGFP)在BEL7402肝癌细胞中的表达变化.     

VEGF反义RNA对人食管癌细胞抑制作用的研究

目的: 研究血管内皮生长因子(VEGF)反义RNA对人食管癌细胞的抑制作用,探讨以VEGF反义RNA进行食管癌基因治疗的可行性.方法: 采用脂质体法将反义VEGFcDNA质粒转染入食管癌细胞TE-1;用MTT法检测细胞增殖情况;用免疫组化法检测VEGF蛋白表达;原位杂交和RT-PCR技术检测VEGFmRNA的表达.流式细胞术检测细胞的凋亡率和周期分布.结果: 被反义VEGFcDNA质粒转染的食管癌细胞有外源性VEGF反义基因的整合及表达,该细胞的VEGFmRNA及蛋白表达水平降低,但细胞生长速率无明显改变,未发生明显凋亡现象;接种裸鼠后,形成肿瘤的时间明显延长,肿瘤的体积和重量明显减小.结论: VEGF反义RNA可抑制食管癌细胞VEGF表达和裸鼠体内的肿瘤生长.该表达体系为食管癌基因治疗的进一步研究奠定了基础.