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Biomedical subjects

L H Wu

Publications and source records attributed to L H Wu.

At least 37 records · Page 2Linked to original sources
PubMed1996

Enhancement of chemosensitivity by tyrphostin AG825 in high-p185(neu) expressing non-small cell lung cancer cells.

The HER-2/neu gene product, p185(neu), is a membrane-bound receptor with tyrosine kinase activity. High levels of p185(neu) is correlated with intrinsic chemoresistance of non-small cell lung cancer (NSCLC) cell lines. We investigated the effects of tyrphostin AG825, a selective tyrosine kinase inhibitor preferentially inhibiting HER-2/neu kinase, on the chemosensitivities and on the drug-induced cell cycle changes of NSCLC cell lines that expressed different levels of p185(neu). Compared to the low-p185(neu) expressing cell lines, we found that the high-p185(neu) expressing cell lines were more resistant to doxorubicin, etoposide, and cis-diamminedichloroplatinum(II) but more sensitive to AG825. AG825 was able to significantly enhance the chemosensitivities of the high-p185(neu) expressing cell lines, whereas it had little effect on the chemosensitivities of the low-p185(neu) expressing cells, with a few exceptions in which minor antagonistic effects were observed. Although high concentrations of AG825 could reduce the drug-induced G(2) arrest that was accompanied by the activation of phosphorylated p34(cdc2), we failed to find any remarkably differential effects of AG825 on drug-induced G(2), arrest and the accompanying phosphorylation status of p34(cdc2) of the high- and and the low-p185(neu) expressing cell lines. In summary, tyrphostin AG825 can enhance chemosensitivity in high- but not in low-p185(neu) expressing NSCLC cell lines. This differential effect cannot be explained by the alterations of drug-induced cell cycle changes by AG825. Our results provide a rationale to develop p185(neu)- specific tyrphostin and to test them in combination with anticancer agents in vivo and in clinical trials.

Benzothiazoles↗
PubMed1996

Correlations between intrinsic chemoresistance and HER-2/neu gene expression, p53 gene mutations, and cell proliferation characteristics in non-small cell lung cancer cell lines.

Using a panel of 20 non-small cell lung cancer (NSCLC) cell lines established from previously untreated patients, we investigated the relationships between intrinsic chemoresistance (to four agents used commonly in the therapy of NSCLC) and HER-2/neu gene expression (which encodes glycoprotein p185neu), p53 gene mutations, and cell proliferation characteristics. Our results demonstrated that high p185neu expression was correlated with chemoresistance, low S-phase fractions, and long doubling times. By contrast, cell lines expressing relatively low levels of p185neu were relatively chemosensitive and had higher S-phase fractions and shorter doubling times. Although mutation of the p53 gene was a common event in this panel of cell lines (present in 18 of 20 lines), there was no relationship between mutations at any specific codon and chemoresistance or cell proliferation characteristics. Multivariate analysis revealed that the level of p185neu was the only independent predictor for chemoresistance to doxorubicin, etoposide, and probably cisplatin. Although intrinsic chemoresistance almost certainly is a multifactorial process, overexpression of p185neu may be an important factor in the chemoresistance of NSCLC.

Carcinoma, Non-Small-Cell Lung↗
PubMed1996

Structural motifs encoded by individual exons of the human neurokinin-1 receptor gene interact differentially with selective agonists and antagonists.

Three chimeric receptors were constructed by exchanging exons between human neurokinin NK1 and NK3 receptor genes. The N-terminal sequences of these chimeric receptors are encoded by exon 1, exon 1-2, or exon 1-3 of the NK1 receptor gene, whereas the remaining C-terminal sequences of these chimeric receptors are encoded by corresponding exons of the human NK3 receptor gene. Substance P bound with high affinities to all three chimeric receptors, suggesting that in addition to the common structures composed of conserved amino acid residues among neurokinin receptors, structural elements encoded by the first exon of the human NK1 receptor gene may also play an important role for substance P binding. On the contrary, potent NK1 antagonists L703,606 and SR140,333 did not show any detectable binding to these chimeric receptors. In accordance, sequences encoded by exon 4, and possibly exon 5, are likely to contain important structural motifs that may directly or indirectly influence the binding of these antagonists. Further comparison of the binding affinities of highly selective NK1 agonists, [Sar9, Met(O2)11] substance P, substance P methyl ester, and septide, revealed that each agonist may interact differently with the human NK1 receptor. These results show that the exon-exchanging technique can be a useful tool for studying structure-function relationships of receptors in which exon-intron junctions are fully conserved among receptor subtypes.

Amino Acid Sequence↗
PubMed1996

DNA flow cytometric analysis of chest-wall chondroma and chondrosarcoma.

DNA flow cytometric analysis was performed for prediction of malignancy potential, and hence of outcome, in 17 patients aged 29-76 (mean 52) years with cartilaginous tumour of the chest wall. Histologically there were nine chondromas and eight chondrosarcomas. The chondrosarcomas (3 grade I, 4 grade II, 1 grade III) were significantly larger than the chondromas (13.6 +/- 7.8 vs 4.2 +/- 2.0 cm, p < 0.001). DNA flow cytometry was performed on paraffin-embedded blocks of resected tumour. In eight of the 17 tumours (6 chondromas, 1 grade I and 1 grade II chondrosarcoma) measurement of DNA content failed because of low cellularity and severe calcification. Aneuploid DNA content was detected only in three chondrosarcomas (2 grade II, 1 grade III). The synthetic phase fraction was less than 15% in all the studied chondromas, but exceeded 25% in four of six chondrosarcomas. This case series though small, may indicate a trend towards association between higher histologic grading and greater incidence of aneuploidy with higher synthetic phase fraction. Tumour size, histologic grade, DNA ploidy and adequacy of surgical resection are the main influences on prognosis.

Adult↗
PubMed1996

Establishment and characterization of two cell lines derived from a single hepatocellular carcinoma containing multiploid DNA distribution.

Previous studies showed that patients with resectable multiploid hepatocellular carcinoma (HCC) had lower overall survival rate and higher recurrent rate than did those with diploid or single aneuploid tumors after hepatic resection. We describe in this study the establishment and characterization of cell lines derived from a single HCC nodule containing multiploid DNA distribution. Two human HCC cell lines, designated HAGS 2.1 and HAGS 2.2, were established by primary culture and single cell cloning methods from a patient with a multiploid HCC tumor. Both cell lines expressed bile canalicular domain-specific antigen of human hepatocyte. The HAGS 2.1 cells were spindle-shaped without prominent intracellular vesicles and had a doubling time of 38 h with DNA ploidy of 4.4 N; cells of HAGS 2.2 were polygonal with many intracellular vesicles and had a doubling time of around 42 h with DNA ploidy of 5.1 N. Hepatitis B surface antigen was detectable in HAGS 2.2 but negative in HAGS 2.1 cells. Both HAGS 2.1 and HAGS 2.2 expressed major histocompatibility complex (MHC) class I antigen, but the former expressed more MHC class II antigen than did the latter. Polymerase chain reaction and subsequent single strain conformation polymorphism analysis disclosed the presence of preS and S regions and the absence of C and X regions of HBV genome in cells of both HAGS 2.1 and HAGS 2.2. We conclude that the establishment of cell lines derived from a single HCC tumor containing multiploid DNA distribution might provide a good in vitro model to study the carcinogenesis and the recurrence mechanism of human hepatocellular carcinoma.

Antigens, Neoplasm↗
PubMed1995

Identification and characterization of c-erbB-2 proteins in serum, breast tumor tissue, and SK-BR-3 cell line.

We have identified and characterized c-erbB-2 protein molecules in sera from patients with carcinomas, in both cytosol and cell membrane extract from breast tumor tissue and in both the culture medium and cell extract of the SK-BR-3 cell line. These proteins were characterized by various chromatographic techniques and identified by the use of two immunoassays; one measures both the c-erbB-2 oncoprotein (p185) and its ectodomain (p120), and the other in-house assay reacts specifically for p185. We found that the majority of the immunoreactivity detected in the serum, tumor tissue cytosol, and conditioned cell medium was derived from the ectodomain molecule (p120) of the c-erbB-2 oncoprotein (p185), whereas only p185 was detected in the extracts from cell membrane of both tumor tissue and the SK-BR-3 cell line. The ectodomain molecules (p120) found in the serum, cytosol, and cell medium were very similar in terms of molecular size and charge property. The molecular weight was determined to be 120 kDa by the size exclusion HPLC method. Both p120 and p185 are glycoproteins and were retained by the ConA Sepharose column. Both molecules are also heterogeneous in charge and multiple peaks could be identified in the elution profiles of anion exchange HPLC and chromatofocusing. This information should not only facilitate the isolation of these molecules, but also improve preparation of specific antibodies, preparation of calibrators, and development of improved assays for these proteins.

Breast Neoplasms↗
PubMed1995

Two classes of structurally different antagonists display similar species preference for the human tachykinin neurokinin3 receptor.

Two classes of structurally different tachykinin neurokinin3 (NK3) antagonists were used to evaluate species difference in antagonist binding between human and rat NK3 receptors. In competition binding experiments with [125I-MePhe7]NKB as radioligand, PD 154740, PD 157672, SR 48968, and SR 142801 displayed lower Ki values for the human NK3 receptor (40 +/- 4, 12 +/- 1,350 +/- 50, and 0.40 +/- 0.05 nM, respectively) than for the rat NK3 receptor (2450 +/- 130, 288 +/- 25, > 10,000, and 11.0 +/- 0.5 nM, respectively). Data from in vitro functional assay showed similar species preference as observed with the competition binding assay. It was shown previously that substitution of only two amino acid residues in the rat receptor to their human counterparts could change the species selectivity of SR 48968, a weak NK3 antagonist. In the double-substituted rat mutant, all three antagonists (PD 154740, PD 157672, and SR 142801) displayed Ki values (76 +/- 8, 16 +/- 2, and 0.50 +/- 0.05 nM, respectively) very similar to the Ki values for the wild-type human NK3 receptor. Thus, in addition to their previously reported effects on SR 48968, these two amino acid residues are responsible for the species selectivity of these three additional NK3 antagonists. Because PD 154740 and PD 157672 are very different structurally from SR 48968 and SR 142801, our results indicate that the two identified residues may be involved in adopting a receptor conformation that favors the binding of NK3 antagonists that display species preference for the human NK3 receptor.

Animals↗
PubMed1994

Identification of methionine134 and alanine146 in the second transmembrane segment of the human tachykinin NK3 receptor as reduces involved in species-selective binding to SR 48968.

We have shown that SR 48968 possess sub-micromolar affinity for the human tachyknin NK3 receptor; however, its affinity for the rat NK3 receptor is greater than 10 microM. To determine the functional domain(s) responsible for the species variation in binding affinities, we have constructed several human/rat chimeric NK3 receptors. Based on studies of these chimeric receptors, the species-specific binding sites for SR 48968 were localized to five residues in the 1st and 2nd transmembrane segments of the human NK3 receptor. We have individually mutated all five residues in the rat receptor to their corresponding residues in human. Only two single-substituted mutants (V121M and G133A) show a small increase in their binding affinities for SR 48968. However, a mutant containing both substitutions was shown to have the same affinity for SR 48968 as the wild type human NK3 receptor. It is concluded that collectively these two amino acid changes are responsible for the species difference in binding affinities for SR 48968.

Alanine↗
PubMed1994

The non-peptide tachykinin NK2 receptor antagonist SR 48968 interacts with human, but not rat, cloned tachykinin NK3 receptors.

SR 48968, a non-peptide tachykinin NK2 receptor antagonist, has been shown to possess sub-micromolar affinity for NK3 receptors present in the guinea pig. In the present study, we have compared the binding affinities of SR 48968 to the cloned human and rat NK3 receptors expressed in CHO cells. Using [125I]-[MePhe7]-neurokinin B as the radioligand, SR48968 displayed an IC50 value of 350 nM for the human NK3 receptor as compared with a value of greater than 10 microM for the rat NK3 receptor. Exposure of cells transfected with human NK3 receptor cDNA to [Pro7]-neurokinin B increased inositol phospholipid turnover in a concentration-dependent manner and this response was blocked competitively by SR 48968. Our results demonstrate that SR 48968 is an antagonist at the human NK3 receptor and may be a useful tool for elucidating the species-dependent variations in the non-peptide antagonist binding site(s) on the NK3 receptor.

Animals↗
PubMed1994

The random inactivation of the X chromosome carrying the defective gene responsible for X-linked hyper IgM syndrome (X-HIM) in female carriers of HIGM1.

The molecular origin of X-linked hyper IgM syndrome has recently been identified as a defect in the ligand of CD40, gp39, a protein expressed on the surface of activated T cells. The availability of detailed pedigrees for three families with affected males allowed assessment of the random or nonrandom nature of the inactivation of the defective X chromosome as well as a determination of the origin of the mutation. X chromosome inactivation was studied because of the relevance to the ability to detect carriers of HIGM1 and the potential for phenotypic effect in the carriers. Using immunostaining, PCR, and DNA sequencing, we found that the defective gene for gp39 is not selectively inactivated. Even in the presence of extremely skewed inactivation, normal levels of serum Ig were found. In carriers in which the defective gene is predominantly expressed, staining alone revealed the carrier status reliably while cloning and sequencing of the cDNA was necessary when the normal gene was predominantly expressed. Unlike some other X-linked defects where extreme Lyonization may lead to disease, a small population of cells expressing the wild-type gp39 is sufficient to maintain normal humoral immunity and prevent the clinical symptoms of X-HIM.

Adolescent↗
PubMed1994

[Pharmacodynamics and clinical therapeutic effects of aerosol and injection of shuanghuanglian].

The results show that both aerosol and injection of shuanghuanglian can increase the activity of NK cells in peripheral blood of the human body as well as the ability of lymphocyte to produce alpha-interferon. Their broad-spectrum antiseptic and virus-inhibiting actions have also been confirmed. The absolute bioavailability of aerosol was 96.88% (baicalin) and 90.69% (chlorogeniacid). The significant response rate of aerosol and injection was 84% and 83% respectively. The total response rate was 92% and 93% respectively.

Adolescent↗
PubMed1994

The molecular basis of X-linked agammaglobulinemia, hyper-IgM syndrome, and severe combined immunodeficiency in humans.

The molecular basis for X-linked agammaglobulinemia, hyper-IgM syndrome, and severe combined immunodeficiency was recently identified. In X-linked agammaglobulinemia the molecular defect was found to reside in the gene encoding a novel cytoplasmic tyrosine kinase (bpk, atk, or btk) expressed by B and myeloid cells. This kinase belongs to a new subfamily of tyrosine kinases that contains SH1, SH2, and SH3 domains. A defect in the murine homologue of this kinase has been shown to be responsible for X-linked immunodeficiency in mice. Currently, the role of btk in B- and myeloid cell signaling is unknown. The molecular defect in X-linked hyper-IgM syndrome has been shown to reside in the gene encoding the T-cell activation protein gp39 (CD40L, TRAP). This protein binds to its counter receptor, CD40, on B cells and has been shown to participate in T-cell-dependent B-cell help leading to B-cell proliferation and isotype switching. X-linked severe combined immunodeficiency patients were found to have defects in the gene encoding the gamma-chain of the interleukin-2 receptor. This chain of the interleukin-2 receptor is constitutively expressed by T cells and is involved in the formation of high and intermediate affinity interleukin-2 receptor complexes. These two interleukin-2 receptor complexes are responsible for mediating interleukin-2-dependent signals.

Agammaglobulinemia↗
PubMed1993

Morphometric and immunocytochemical assessment of fungiform taste buds after interruption of the chorda-lingual nerve.

Unilateral interruption of the chorda-lingual nerve led to a loss of most epithelial axons and to the deterioration of fungiform taste buds in the anterior portion of the tongue of albino rats, mongolian gerbils, and golden hamsters. By three weeks after surgery the following percentages of fungiform taste buds had completely disappeared: 71% in gerbils, 28% in rats, and 26% in hamsters. Residual taste buds were classified into two groups: atrophic taste buds and taste bud remnants. Atrophic taste buds were smaller than normal and typically had no visible taste pore, although they retained the characteristic oval shape of a taste bud and numerous elongated cells. Taste bud remnants were non-oval fragments of taste buds with few elongated cells. Specific markers for elongated taste cells (monoclonal antibodies to keratin 19) confirmed that atrophic taste buds, as well as some taste bud remnants, had elongated taste cells. By 180 days after chorda-lingual nerve transection, 44% of rat fungiform taste buds had disappeared; morphometric analysis of the 311 residual taste buds established that 241 atrophic taste buds and 69 taste bud remnants were, respectively, 50% and 75% smaller than the average volume of 480 normal taste buds. The aggregate loss of gustatory tissue, calculated from the shrinkage of residual taste buds and the volume lost by the outright disappearance of many taste buds, was 88% for gerbils, 72% for rats, and 65% for hamsters. Evaluation in gerbils of the co-occurrence of taste buds and axons suggests residual taste buds were neurotrophically supported. Every gerbil fungiform papilla that lacked axons lacked a taste bud.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

DNA ploidy as a major prognostic factor in resectable ampulla of Vater cancers.

The archival paraffin-embedded specimens from 63 ampulla of Vater cancers after pancreaticoduodenectomy between 1965 and 1991 were analyzed by flow cytometry. Of the 63 cancers, 31 (49.2%) were diploid DNA cancers and 32 (50.8%) were aneuploid. Patients with diploid DNA cancer had a median survival time of 159.0 months, and patients with aneuploid DNA cancer had 24.0 months. This difference is statistically significant (P = 0.0257). The aneuploid group did have a poorer prognosis than the diploid group. The multivariate analysis demonstrated that DNA ploidy was an independent and very important prognostic factor, even stronger than the stage and lymph node status. There was a tendency toward higher values of S-phase fraction, proliferative index, and total aneuploid DNA fraction in the shorter survival groups, but they were of no statistical significance. These data suggest that DNA ploidy appears to be the most important and the only pre-operative predictor of prognosis in resectable ampulla of Vater cancers since endoscopic biopsy is feasible.

Ampulla of Vater↗
PubMed1993

Suicide attempts in rapid cycling bipolar disorder patients.

We studied 100 rapid cycling patients (33% of whom had truncated episodes), and 120 non-rapid cycling bipolar patients for history of suicide attempts. In contrast to our hypothesis, no significant differences were found when comparing suicide attempt histories for these two groups.

Adult↗
PubMed1993

Deoxyribonucleic acid histogram of testes in primary transsexualism.

The influence of oestrogen on spermatogenesis was evaluated by comparing its effect on the testes of primary male transsexuals and those who had undergone hormone therapy for prostatic carcinoma. Eight primary transsexuals were studied. They had been diagnosed on clinical and psychiatric evidence and had been on oestrogen therapy for several years since puberty. Histological sections of testicular tissue obtained at reassignment surgery from 8 phenotypic male transsexuals (aged 24-32 years) with an XY chromosome complement were studied by light microscopy. The formalin-embedded specimens were analysed by flow cytometry for deoxyribonucleic acid (DNA) histograms. Both the histology and DNA histograms revealed a pattern of maturation arrest in 12 of 16 testes in which the diploid cell compartment occupied most of the spermatogenetic element, followed by tetraploid and monoploid cells. Two testes showed impaired spermatogenesis and 2 were normal. The DNA histograms and pathology were also evaluated in 20 testes after 3 to 8 years of hormone therapy in patients with advanced prostatic carcinoma (aged 60-78 years). No maturation arrest was found in these patients. Sixteen of them had a pattern of fibrosis and atrophy and 4 had impaired spermatogenesis. It was concluded that oestrogen influenced spermatogenesis and affected the maturation of spermatogonia mostly during puberty.

Adult↗