人原发性肝癌中的转甲状腺素蛋白(Transthyretin)基因

本文从人正常肝对原发性肝癌的递减式cDNA文库(Subtracting cDNA libra-ry)中筛选出pG8cDNA克隆.Northem杂交证明它在正常肝中高度转录,而在9例肝癌中转录严重受阻遏.9例肝癌DNA MspI酶切杂交信号提示,4例肝癌中该基因存在部分DNA片段的丢失.在另7对肝癌及癌旁组织样本中,有4例肝癌中该基因同样存在部分片段丢失.cDNA序列分析证明它与转甲状腺素蛋白(Transthyretin,TTR)基因的编码区全部同源.本文首次报道了在人肝癌中TTR基因转录严重受阻遏及在基因结构上可能存在丢失或缺陷,提示TTR基因可能是人肝癌中基因缺陷的一个标记或抗癌基因之一.     

IDENTIFICATION OF HUMAN N-ras AS THE COMMON ONCOGENE IN NIH 3T3 CELLS TRANSFORMED WITH DNAs FROM HUMAN PRIMARY HEPATIC CANCER AND HEPATOMA 7402 LINE

DNA was extracted from NIH 3T3 cells transformed with DNAs from human primary hepatic cancer (PHC) and Hepatoma 7402 cell line. The transformant DNA was analyzed by Southern transfer and hybridization with ~(32)P-labeled probes of various oncogenes.The EcoRI 7.2 and 9.0 kb bands characteristic of human N-ras gene were identified in transformed NIH 3T3 cells derived both from PHC and 7402 DNA. The BamHI 6.6kb band characteristic of human c-Ha-ras I was present only in 7402 transformants, but not in PHC transformants.Using ~(35)S-methionine incorporation, immunoprecipitation with anti-p21 monoclonal antibodies, SDS-PAGE and autoradiography, it was demonstrated that p21 synthesis was remarkably enhanced in 1402 cells as well as in transformed cells derived from both 7402 and PHC DNA.Taking the data together, it. strongly implies that N-ras is one of the transforming genes for human liver cancer.     

鸭肝癌癌基因的研究——mht(raf)及Ha-ras

本文首次报道了鸭肝癌及癌旁组织DNA对NIH 3T3及RAT-1细胞的转化活性,并获得了转化细胞株。从转化细胞株DNA分析,证明鸭肝癌和癌旁DNA的转化细胞株中均存在mht(mil或raf)和C-Ha-ras的顺序。这对进一步研究乙型肝炎病毒,化学致癌物引发鸭肝癌的分子机理提供了重要的基础。     

TRANSFORMING GENES IN DUCK HEPATIC CARCINOMA——mht(raf) AND Ha-ras

The transfection of NIH 3T3 cells was performed with DNAs from 2 duck primary hepatic carcinomas (DHC 40K, 9K) and I tumor-adjacent liver tissue (TAL, 9N). Transfectants were found from 40K, 9K and 9N DNAs. The secondary transfectants were obtained after transfection of RAT-1 cells with DNAs from primary transfectants. After hybridization with Ha-ras, Ki-ras, N-ras and mht oncogenes, it was found that duck mht (5.2 and 3.2 kb EcoRI fragments) and duck Ha-ras (3.4 kb EcoRI fragments) were present in all these transformants.This is the first report on transforming genes in duck primary hepatic cancer as well as tumoradjacent liver tissue     

TRANSTHYRETIN (PREALBUMIN) GENE IN HUMAN PRIMARY HEPATIC CANCER

From a subtracting cDNA library constructed from normal liver versus human primary hepatic cancer (PHC), a cDNA clone pG8 was isolated. Using it as a probe, RNA extracted from one human liver and 9 PHC samples were analyzed by Northern hybridization. As expected, its mRNA was highly expressed in liver; however, the expression was strikingly suppressed in PHC. Only weak signal was observed in 2 out of 9 PHC, while no signal was detectable in the other 7 samples. Utilizing pG8 as a probe, DNA from the same PHC specimens was analyzed after MspI digestion and Southern hybridization. Deletion of DNA fragment was observed in 4 out of 9 samples. In further study of cancer and non-cancerous liver from other 7 PHC patients, similar deletion of DNA fragments in cancer was observed in 4 out of 7 samples. After sequencing of the clone of 572 bp, it was unexpectedly found that pG8 was completely homologous to the coding sequence of transthyretin, TTR gene, as TTR (or prealbumin) gene has been known to be linked to a     

人原发性肝癌及7402细胞株和髓细胞白血病细胞株(K562)的癌基因(转化基因)的共同属性——N-ras基因

用人的原发性肝癌(PHC)、肝癌7402细胞株(郭婵等,1984)及髓细胞白血病K562细胞株的DNA转染的NIH/3T3转化株DNA,以Southern吸印和~(32)P-标记的癌基因探针做分子杂交来分析,结果如下:在PHC DNA转化的NIH/3T3细胞DNA中,鉴定有N-ras基因的7.2及9.0kb(EcoRI酶切)的带,与人白细胞和肝癌DNA中出现的专一性条带相同,但是没有人Ha-ras(BamHI 6.6kb)或Ki-ras(EcoRI 3.0kb)的专一性条带。在以7402及K562细胞株DNA转化的细胞DNA中,同时发现有人的N-ras和Ha-ras基因(6.6kb,BamHI酶切),人的Ki-ras特异片段(3.0kb,EcoRI酶切)没有被检测到。PHC的DNA中N-ras基因在带型及杂交强度上没有大的改变,由于N-ras基因(非Ki-ras,Ha-ras)的表达,在多数PHC中明显地增加,提示了N-ras至少是人肝癌的转化基因之一。     

Nucleotide Sequence of a Cloned Human HBV Mutant (pDKHBV) in Duck Hepatoma of Qidong County

This is the first report to describe the presence of an HBV-like DNA sequence in two hepatoma and their tumor surrounding liver tissues, one precancerous and one non-malignant liver tissue of ducks collected from Qidong County of China. The HBV-like sequences were either in an episomal form of 3.2 kb or in an integrated form of various sizes, while the DHBV DNA sequences (3.0 kb) were either present or absent in these tissues and in different size pattern. Furthermore, there was no evidence of cross-hybridization between HBV-like DNA sequences in duck and DHBV DNA. A 3.2 kb HBV-like DNA sequence has been cloned from one duck hepatoma (40 K), designated as pDKHBV. The 3218 bp full-length nucleotide sequence of this clone has been determined, which had no apparent homology with Duck Hepatitis B Virus (DHBV) genome, but was highly homologous to human HBV adw_2 subtype (99.0%). The sequence was composed of four open reading frames for HBV gene Pre-S/S, X, C and P respectively. In addition to multiple sites     

启东鸭肝癌中人HBV突变体(pDKHBV)的DNA序列

本文首次报道对启东鸭肝癌中存在的人HBV样DNA~([3])(pDKHBV)的全序列分析。本工作建立了pDKHBV的M_(13)重组克隆及亚克隆,并应用末端终止法测定了它的全序列。结果表明,它长3218bp,与鸭乙型肝炎病毒无关,而同人乙型肝炎病毒(HBV)基因组有91％以上同源性(其中与adw_2亚型有99.0％同源性),存在与HBV一致的4个开放阅读框架,并在Pre-S_1基因下游59个核苷酸,以及C基因3′端分别有1个和2个核苷酸的缺失,造成Pre-S_1基因第58个密码子、C基因第158个密码子出现终止信号,以及P基因第21个密码子起发生移码突变。以上结果表明启东鸭肝癌中存在adw_2亚型HBV的非复制型突变体。由于启东鸭肝癌高发且与人肝癌高发有相关性,提示HBV作为人和鸭肝癌共同病因的可能性。