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L L Hsieh

Publications and source records attributed to L L Hsieh.

66 records · Page 4Linked to original sources
PubMed1988

Immunological detection of aflatoxin B1-DNA adducts formed in vivo.

Two monoclonal antibodies (6A10 and 12F5) were obtained after fusion of mouse P3X63-AG.8.653 myeloma cells with spleen cells isolated from BALB/c mice immunized with imidazole ring-opened aflatoxin B1 (AFB1)-DNA and characterized by competitive enzyme-linked immunosorbent assays. Both antibodies are highly specific for imidazole ring opened AFB1-DNA and show some cross-reactivity with AFB1-DNA and no cross-reactivity with 8,9-dihydro-8-(7-guanyl)-9-hydroxy-AFB1, AFB1 conjugated with bovine serum albumin, aflatoxin M1 conjugated with bovine serum albumin, AFB1, or aflatoxin G1. Antibody 6A10 was further characterized and showed no cross-reactivity with DNA modified by several other carcinogens. It could detect adducts with 4-fold higher sensitivity in highly modified DNA (2.5 adducts/100 nucleotides) than in low modified DNA (4 adducts/10(5) nucleotides). With low modified DNA the limit of sensitivity is 5 adducts/10(7) nucleotides. Antibody 6A10 reliably detected adducts formed in vivo in rats and mice treated with AFB1. In a pilot study, AFB1 adducts were detected in liver tissues from individuals living in areas with suspected exposure to AFB1. Monitoring adduct levels in human tissue may provide information not only on carcinogen exposure but also on the relationship among infection with hepatitis B virus, dietary exposure to aflatoxin B1, and liver cancer.

Aflatoxin B1↗
PubMed1988

Amplification of the EGF receptor and c-myc genes in human esophageal cancers.

The incidence of esophageal cancer is extremely high in Linxian County and certain other regions of the People's Republic of China. Epidemiologic and laboratory studies suggest that N-nitroso carcinogens and other environmental factors play a causative role. In the present study, employing over 100 DNA samples obtained from Lin-xian patients who underwent surgery for esophageal cancer, we have found a significant frequency of amplification of either the human epidermal growth factor receptor (HER-I) gene or the c-myc oncogene. These changes were found not only in tumor specimens, but also in adjacent non-tumor (grossly normal) tissue specimens obtained from patients with esophageal cancer. RNA samples were also obtained from over 30 tissue samples. These revealed considerable variation in the abundance of HER-I and c-myc transcripts in both the tumor and adjacent non-tumor specimens. A few samples revealed extremely high levels of these transcripts. Thus, changes in gene copy number or level of expression of HER-I or c-myc DNA sequences may play an important role in the pathogenesis of esophageal cancer in this high-risk region.

Blotting, Northern↗
PubMed1988

Changes in expression of oncogenes and endogenous retroviral-like sequences during colon carcinogenesis.

The possible roles in experimental colon carcinogenesis of two protooncogenes (c-myc and c-H-ras), two endogenous retrovirus-related DNA sequences [rat leukemia virus (RaLV) and the 30S sequence], and two cell cycle related genes (beta-actin and ornithine decarboxylase) were studied by analyzing the levels of their corresponding RNAs during the course of azoxymethane induced and high fat promoted colon carcinogenesis. F-344 male rats received three s.c. injections of azoxymethane (15 mg/kg) or normal saline and were then subdivided into high or low fat diet groups. During subsequent serial sacrifices normal colon mucosa, adenomas, and carcinomas were harvested for histology and RNA extraction. Seventy-one RNA samples were analyzed by the Northern blot hybridization procedure using the appropriate 32P-labeled DNA probes. A marked increase in the abundance of c-myc, RaLV, and 30S RNAs were seen in all of the colon tumors, including adenomas and invasive carcinomas. No or a very low level of expression of RaLV and c-myc RNA was found in the flat grossly normal mucosa adjacent to the tumors and in the mucosa of the control rats. Some of the colon tumors also displayed increased levels of c-H-ras, ornithine decarboxylase and beta-actin RNAs but these findings were less striking and more variable than those seen with c-myc, RaLV, and 30S RNAs. These results suggest that increased expression of the c-myc protooncogene and of the endogenous retrovirus-like sequences (RaLV) and 30S are hallmarks of colon carcinogenesis in this model system.

Animals↗
PubMed1988

Expression of endogenous retrovirus-like sequences and cellular oncogenes during phenobarbital treatment and regeneration in rat liver.

The expression of two cellular oncogenes (c-myc and c-Ha-ras), the epidermal growth factor receptor gene, and two endogenous retrovirus-like sequences (rat leukemia virus and 30S) was examined in control (nonregenerating) rat livers and at various times after partial hepatectomy. One group of rats had been fed phenobarbital (0.05%) for 16 days prior to the partial hepatectomy. The feeding of phenobarbital (0.05%) itself led to a 65% decrease in the level of epidermal growth factor receptor RNA, but no major change in the level of c-myc, H-ras, rat leukemia virus, or 30S RNAs, in the control rat livers. There was a considerable increase (4- to 5-fold) in the level of c-myc transcripts, at 12 and 48 h after partial hepatectomy in the phenobarbital-treated rats, and at 12 and 24 h in the rats on the control diet. By 72 h, the level of c-myc transcripts returned to normal in both groups of rats. A slight increase (about 1.5-fold) in the level of c-H-ras transcripts was seen at 24 h, which returned to normal levels by 168 h, in the regenerating livers of both the phenobarbital-treated and control diet rats. The regenerating livers displayed a marked decrease (3- to 4-fold) in the level of epidermal growth factor receptor RNA in both the phenobarbital and control diet rats. A marked increase (5- to 6-fold) in the level of transcripts homologous to the endogenous rat leukemia virus-like sequence was seen at 24 h in all of the regenerating livers, but there was no significant change in the level of RNAs homologous to 30S. Thus, the proliferation of normal rat liver cells mimics some but not all of the changes in mRNA levels that we have previously described in rat liver tumors.

Animals↗
PubMed1987

Expression of retroviral sequences and oncogenes in rat liver tumors induced by diethylnitrosamine.

The expression of three cellular oncogenes (c-myc, c-Ha-ras, and c-delta-raf), the epidermal growth factor receptor gene, and two endogenous retrovirus-like sequences [rat leukemia virus (RaLV) and 30S] was examined in control rat livers and in 16 liver tumors. The tumors were induced in Sprague-Dawley male and female rats by a single i.p. injection of diethylnitrosamine at 1 or 2 days after birth, followed by dietary exposure to phenobarbital beginning at weaning. Increased expression of c-myc was seen in most of the tumors, but there was no consistent increase or decrease in expression of c-Ha-ras or c-delta-raf. It is of interest that a number of the tumor samples showed a decrease in epidermal growth factor receptor RNA. In all of the tumors, including both hepatocellular adenomas and carcinomas, there was a marked increase in expression of the endogenous RaLV sequence, and over 90% of the tumors displayed increased expression of the 30S endogenous retroviral-like sequence. No or a very low level of expression of the RaLV and 30S sequences was found in the control livers. The extent of expression of the RaLV and 30S sequences in individual tumors did not correlate with the extent of expression of c-myc or c-Ha-ras. Although increased expression of certain endogenous retrovirus-related sequences appears to be a common finding during rat liver carcinogenesis, the significance of this finding remains to be determined.

Animals↗
PubMed1986

Immunologic quantification of carcinogen-DNA adducts.

Sensitive immunological methods for the detection of carcinogen-DNA adducts have recently been developed. These techniques are particularly useful for screening human populations for exposure to environmental carcinogens. Measurement of the biologically effective dose in humans may be useful in detecting carcinogenic hazards and carrying out risk estimates. We have developed monoclonal antibodies to several carcinogen-DNA adducts. These have included DNA modified by a benzo[a]pyrene diol epoxide (BPDE-I), 1-aminopyrene (1-AP) and 8-methoxypsoralen (8-MOP). BALB/cCr mice were immunized with the modified DNAs complexed electrostatically to methylated bovine serum albumin. Several stable clones have been isolated for each of the modified DNAs and characterized by enzyme-linked immunosorbent assay (ELISA). All antibodies are highly specific for the appropriate modified DNA and do not cross-react with nonmodified DNA. The antibodies to BPDE-I-DNA have significant cross-reactivity with DNAs modified by similar antitrans diol epoxides of benz[a]anthracene and chrysene. These DNAs all contain N-2 of guanine adducts. The antibody probably recognizes a shared determinant encompassing the guanine base and the hydrocarbon ring containing the hydroxide groups. The antibodies cross-react with BPDE-I-dG, the monoadduct isolated from DNA, but with lower sensitivity than for the intact modified DNA. They do not react with acetylaminofluorene (AAF) or 1-AP modified DNA, both of which contain C-8 of guanosine adducts. The antibodies to 1-AP-modified DNA demonstrate cross-reactivity with 8-nitro-1-aminopyrene- and 6-nitro-1-aminopyrene-modified DNA, as well as some slight cross-reactivity with BPDE-I-DNA and AAF-DNA.(ABSTRACT TRUNCATED AT 250 WORDS)

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1985

Monoclonal antibodies to 1-aminopyrene-DNA.

Monoclonal antibodies were obtained after fusion of mouse P3X63-AG.8.653 myeloma cells with spleen cells isolated from BALB/cCr mice immunized with denatured DNA modified by 1-nitrosopyrene reduced with sodium ascorbate (AP-d-DNA) and complexed electrostatically to methylated bovine serum albumin. Ten stable hybridoma lines have been isolated and characterized by enzyme-linked immunosorbent assay (ELISA). They all recognize 1-aminopyrene (1-AP)-modified DNA, but not free 1-nitropyrene or 1-aminopyrene. Antibody 11H2 is the most specific for AP-DNA showing no cross-reactivity with unmodified native DNA. It also recognizes 8-nitro-1-AP and 6-nitro-1-AP modified DNA. There was some low cross-reactivity with DNA modified by a benzo[a]pyrene diol epoxide and N-acetoxy-N-2-acetylaminofluorene. Competitive ELISA with antibody 11H2 reliably detected AP-DNA adducts formed when 1-nitropyrene was incubated with Salmonella typhimurium TA1538. By immunological methods, AP-DNA adducts were shown to be unstable to heat denaturation. This suggests that specific monoclonal antibodies to carcinogen-DNA adducts will be useful not only for detecting and quantitating carcinogen-DNA damage but also for probing adduct stability.

Aminopyrine↗
PubMed1985

Quantitation of exposure to benzo[a]pyrene with monoclonal antibodies.

It is now possible to quantitate carcinogen adducts on DNA by highly sensitive immunoassays. These techniques are particularly useful for screening human populations for exposure to potential environmental carcinogens. We have developed a panel of monoclonal antibodies that react with benzo(a)pyrene (BP) modified DNA to be used in an enzyme linked immunoassay (ELISA) to quantitate adduct levels of both human and animal samples. BALBc/Cr mice were immunized with either DNA modified by 7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9, 10-tetrahydrobenzo(a)pyrene (BPDE-I-DNA) complexed electrostatically to methylated bovine serum albumin or with BPDE-I-modified guanosine conjugated with bovine serum albumin (BPDE-I-G-BSA). Four stable clones were produced from the spleen cells of animals immunized with BPDE-I-DNA and one from BPDE-I-G-BSA immunized animals. All antibodies were shown to be highly specific for BPDE-I-DNA and did not crossreact with nonmodified DNA or with N-2-acetylaminofluorene or 1-aminopyrene modified DNA. The antibodies differed in their sensitivity to BPDE-II-DNA, BPDE-I-poly G, BPDE-I-tetraols and BPDE-I-dG. In general, all the antibodies showed the greatest affinity for their original antigen. Those generated against modified DNA showed highest reactivity against modified DNA while the one antibody generated against the monoadduct showed highest reactivity with the monoadduct. These antibodies are currently being used in a highly sensitive competitive ELISA to quantitate levels of BP-DNA adducts in various animal and human tissue samples.

Animals↗
PubMedDate not supplied

p16INK4 gene mutation and allelic loss of chromosome 9p21-22 in Taiwanese hepatocellular carcinoma.

BACKGROUND: The p16INK4 (MTS1/CDNK2A) gene, located on chromosome 9p21, is an inhibitor of cyclin-dependent kinase 4. Various data have shown that it is frequently inactivated in several types of cell lines and primary human cancers. MATERIALS AND METHODS: Thirty cases with hepatocellular carcinoma were studied for possible p16INK4 gene mutation in Taiwan. Homozygous deletion was determined using polymerase chain reaction (PCR). The p16INK4 gene mutation was first screened by single strand conformation polymorphism, then direct DNA sequencing was performed on the cases with mobility shifts. Deletion mapping of chromosome 9p21-22 was also carried out with two polymorphic microsatellite markers (D9S925 and D9S168) using PCR. RESULTS: One of the 30 cases had homozygous deletion at exon 3 of the p16INK4 gene. Another tumor had altered electrophoresed mobility in exon 2 with G to T transversion in the first nucleotide of codon 61 by direct sequencing causing a stop codon (GAG-->TAG). At the D9S925 and D9S168 loci, six out of 24 (25%) and three out of 19 (16%) informative cases showed loss of heterozygosity, respectively. CONCLUSION: Point mutation and homozygous deletion of the p16INK4 gene are present in a subset of hepatocellular carcinomas in Taiwan. The patterns of the p16INK4 gene alteration are, however, different from those from other regions. In addition, allelic loss on chromosome 9p21-22 is not an uncommon event in hepatocellular carcinomas. Therefore, the significance of chromosome 9p loss deserves to be extensively investigated.

Adult↗