Adenovirus-mediated gene therapy specific for small cell lung cancer cells using a Myc-Max binding motif

Recent clinical trials of gene therapy for patients with thoracic cancers have shown that these treatments were well tolerated with minimal side effects and that we need to further enhance specificity as well as efficiency of gene transfer to target cancer cells. We previously reported that myc-overexpressing SCLC cell lines became selectively sensitive to ganciclovir (GCV) by transducing the herpes simplex virus thymidine kinase (HSV-TK) gene under the control of the Myc-Max response elements (a core nucleotide sequence, CACGTG) and that this construct (MycTK) could be utilized to develop a novel treatment against chemo-radio-resistant SCLC. We report here in vivo antitumor effects and safety of a replication-deficient adenoviral vector containing the Myc-Max binding motif (AdMycTK) on SCLC cells. In vitro infection with AdMycTK selectively rendered myc-overexpressing SCLC cell lines 63- to 307-fold more sensitive to GCV. In vivo injections with AdMycTK followed by GCV administration markedly suppressed the growth of myc-overexpressing tumors established in the subcutis or in the peritoneal cavity of athymic mice. On the other hand, infection with AdMycTK did not significantly affect either in vitro GCV sensitivity of the cells expressing very low levels of the myc genes or the growth of their subcutaneous tumors. Moreover, we observed no apparent side effects of this treatment including body weight loss or biochemical abnormalities in contrast to the treatment with AdCATK that conferred strong but nonspecific expression of the HSV-TK gene. These results suggested that AdMycTK/GCV therapy is effective on SCLC patients whose tumors overexpress myc family oncogenes.     

Catechol estrogens induce oxidative DNA damage and estradiol enhances cell proliferation

Estrogen-induced carcinogenesis involves enhanced cell proliferation (promotion) and genotoxic effects (initiation). To investigate the contribution of estrogens and their metabolites to tumor initiation, we examined DNA damage induced by estradiol and its metabolites, the catechol estrogens 2-hydroxyestradiol (2-OHE(2)) and 4-hydroxyestradiol (4-OHE(2)). In the presence of Cu(II), catechol estrogens formed piperidine-labile sites at thymine and cytosine residues in (32)P 5'-end-labeled DNA fragments and induced the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine. NADH markedly enhanced Cu(II)-dependent DNA damage mediated by nanomolar concentrations of catechol estrogens. Catalase and bathocuproine inhibited the DNA damage, suggesting the involvement of H(2)O(2) and Cu(I). These results suggest that H(2)O(2), generated during Cu(II)-catalyzed autoxidation of catechol estrogens, reacts with Cu(I) to form the Cu(I)-peroxide complex, leading to oxidative DNA damage, and that NADH enhanced DNA damage through the formation of redox cycle. To investigate the role of estrogens and their metabolites in tumor promotion, we examined their effects on proliferation of estrogen-dependent MCF-7 cells. Estradiol enhanced the proliferation of MCF-7 cells at much lower concentrations than catechol estrogens. These findings indicate that catechol estrogens play a role in tumor initiation through oxidative DNA damage, whereas estrogens themselves induce tumor promotion and/or progression by enhancing cell proliferation in estrogen-induced carcinogenesis.     

Characterization of mucin antigens recognized by monoclonal antibodies raised against human colon cancer cells

Two monoclonal antibodies, MLS 102, which recognizes cancer-associated mucin antigens, and MLS 103, which recognizes normal mucin, were used to isolate, by immunoaffinity chromatography, the corresponding antigens from cell lysates and spent medium of a human colorectal carcinoma cell line, LS 180. The MLS 102 antigen contained serine, threonine, and proline as major amino acids. The carbohydrate chains of the MLS 102 antigen were composed of O-linked NeuAc alpha 2----6GalNAc (56%), N-acetylgalactosamine (25%), and longer oligosaccharide chains. The MLS 103 antigen differed from the MLS 102 antigen in both amino acid and carbohydrate composition. Most O-linked oligosaccharides of the MLS 103 antigen were longer than the disaccharide found in the MLS 102 antigen. Immunostaining of LS 180 cells using MLS 102 and MLS 103 revealed that the cells are heterogeneous with respect to the expression of the antigens.     

Effect of methionine-enkephalin analog (FK 33-824) on plasma motilin

In an attempt to elucidate the effect of opioid peptide on the secretion of various gastrointestinal hormones, the methionine-enkephalin analog, FK 33-824 (FK) was injected intramuscularly in healthy male adults. The plasma levels of motilin, gastrin and cholecystokinin were assessed by specific radioimmunoassay. After the injection of FK, the plasma level of motilin markedly decreased from the baseline value of 456 +/- 70.2 pg/ml to 264 +/- 44.7 pg/ml at 120 min. On the other hand, the plasma levels of cholecystokinin and gastrin remained unchanged. These data indicate that endogenous methionine-enkephalin may have an direct inhibitory effect on the secretion of motilin.     

Ion specificity of rat chorda tympani fibers to chemical and electrical tongue stimulations

Responses of rat single chorda tympani fibers to ionic chemical stimuli and anodal tongue stimulations with ionic bathing solutions were examined. Fourteen out of 30 fibers showed the highest sensitivities to NaCl for both chemical and electrical stimulations, whereas the remaining 16 fibers showed almost no ion specificity for electrical stimulation even though most of them have the highest sensitivities to HCl for chemical stimulation. Ion specificity for electrical stimulation was significantly smaller than that for chemical stimulation.     

A Case of Xeroderma Pigmentosum with Clinical Appearance of Dyschromatosis Symmetrica Hereditaria

A 9-year-old boy complained of increased freckling on the extremities since very early infancy. Examination showed mottled pigmentation with areas of depigmentation on the backs of the hands and tops of the feet, and less strikingly on the arms and legs. Scattered, small, pigmented freckles on the face were also noticed. The condition was clinically diagnosed as dyschromatosis symmetrica hereditaria (DSH) at first. Unscheduled DNA synthesis was reduced to 66% of the normal value, however, and an ultraviolet sensitivity test by colony formation on fibroblasts revealed moderate sensitivity. Therefore this case was finally diagnosed as xeroderma pigmentosum. It is necessary to study cellular repair capacity to make a precise diagnosis when the distinction is so difficult.     

Selective projections of cholecystokinin-8 immunoreactive fibers to galanin immunoreactive sympathetic preganglionic neurons in a teleost, Stephanolepis cirrhifer.

In the cellular column of sympathetic preganglionic neurons (SPNs) of the filefish Stephanolepis cirrhifer, neurons containing galanin (GAL) form a distinct population projecting specifically to non-adrenergic postganglionic neurons in the celiac and cranial sympathetic ganglia. The present study showed that virtually all of the GAL-immunopositive SPNs made contact with many nerve terminals immunopositive for cholecystokinin octapeptide (CCK-8). GAL-negative preganglionic neurons made contact with only 26% of this type of nerve terminal; CCK-8-immunopositive nerve fibers appeared to project selectively to GAL-immunopositive SPNs with projections to specific targets. The CCK-8-positive nerve fibers might be of primary sensory origin, and participate in the visceral reflexes.