Why do most primary bladder neoplasms first appear around the ureteric orifices?

The majority of primary bladder neoplasms are known to arise within the mucosa around the ureteric orifices and bladder base. This may be due to the mucosa in this area being more susceptible to carcinogens than other areas of the bladder. Deficiency in the nucleotide salvage pathway enzyme thymidine kinase (TK), and especially its TK1 isozyme, has been shown to predispose cell lines to increased mutagenesis. Total TK and TK1 activities were measured in mucosal samples taken adjacent to the ureteric orifices and dome in 32 normal bladders and both total TK and TK1 were shown to be significantly decreased in the mucosa adjacent to the ureteric orifices. This may explain why primary bladder neoplasms occur more commonly in this site.

Breast tumour thymidine kinase levels and disease recurrence.

Thymidine kinase (TK) exists in two forms, TK1 and TK2. TK levels and oestrogen receptor status (OR) were measured in tumours from 86 patients with operable breast cancer and the patients were monitored for recurrence over 24 months. During the monitored period, 13 patients showed recurrence. These patients also exhibited higher total TK levels per mg tumour (P < 0.01) and higher TK1 levels (P < 0.001) than those who did not show recurrence. TK1 levels relative to TK2 were significantly higher (P < 0.05) in OR-negative tumours (n = 29) than in OR-positive tumours (n = 57). OR-negative (n = 9) and OR-positive (n = 4) patients who recurred had significantly higher TK1 levels relative to TK2 than did those who not recur (n = 20 and n = 53, respectively). These preliminary results indicate that breast tumour TK levels may have value in determining prognosis.

Sensitivity to cell killing and the induction of cytogenetic damage following gamma irradiation in wild-type and thymidine kinase-deficient Friend mouse erythroleukaemia cells.

Wild-type Friend erythroleukaemia (clone 707) cells and 2 thymidine kinase-deficient subclones, 707BUE and 707BUF, having thymidine kinase activities of 1.4% and 0.7% that of clone 707, were compared for sensitivity to killing and the induction of cytogenetic damage following gamma irradiation. Three doses of gamma irradiation were used (150, 300 and 450 cGy), and cells were harvested for metaphase spreads after 4, 8, 12, 15, 29 and 43 h. G2 delay was evident at 4 h following gamma irradiation in the 3 cell clones examined, and recovery of mitosis was observed to be dose-dependent. G2 delay was found to be most prolonged in subclone 707BUE and most prompt in clone 707. Increased sensitivity to the induction of cytogenetic aberrations at all three doses was apparent in the 2 thymidine kinase-deficient subclones (as compared to wild-type cells) at 15, 29 and 43 h. Th thymidine kinase-deficient subclones also showed increased sensitivity to gamma radiation-induced cell killing. Furthermore, subclone 707BUE consistently exhibited greater resistance to gamma irradiation than did the subclone with lower thymidine kinase activity, 707BUF. The importance of thymidine kinase levels and extended G2 delay for DNA repair processes is discussed.

Synergism between U.V. and thymidine treatments in the induction of cytogenetic damage in wild-type Friend erythroleukaemia cells.

The cytogenetic aberration-inducing effect of excess thymidine and U.V. light in Friend erythroleukaemia (clone 707) cells was investigated. Three doses of U.V. were utilised, namely 2.4, 4.8 and 7.2 J/m2. Thymidine, at 1 X 10(-5) M, was present for 48 h prior to U.V. treatment and for 15 h following it. Although no significant increase in metaphase aberrations was observed following thymidine treatment alone, relative to the spontaneous frequency, clone 707 exhibited increased sensitivity to U.V.-induced cytogenetic damage, when grown in the presence of 1 X 10(-5) M thymidine. The observed synergism between U.V. and thymidine treatments may be due to thymidine-induced nucleotide pool imbalance with consequent inaccuracies in DNA repair.

Abilities of wild-type and thymidine kinase-deficient Friend mouse erythroleukemia cells to undergo unscheduled DNA synthesis following mutagen treatment.

The abilities of wild-type and thymidine kinase-deficient Friend mouse erythroleukemia cells to perform unscheduled DNA synthesis (UDS), through the incorporation of [3H]deoxycytidine, were measured following damage with methyl methane sulfonate (MMS), ethyl methane sulfonate (EMS), and ultraviolet (UV) irradiation. For each mutagenic treatment, a positive and quantitatively similar response was observed for both wild-type and thymidine kinase-deficient cells. The extent of the response varied greatly, however, depending upon the mutagen used. The results contrast with the unscheduled incorporation of [3H]thymidine in wild-type cells following mutagen treatment, where less variation between the positive UDS responses elicited by MMS, EMS, and UV treatments was observed. Nevertheless, the results clearly indicate that thymidine kinase deficiency does not prevent excision repair (UDS) from occurring.

Enhanced synergism between caffeine and mitomycin C in the induction of cytogenetic aberrations in thymidine kinase-deficient Friend murine erythroleukaemia cells.

The thymidine kinase-deficient subclone, 707BUF, of the Friend murine leukaemia cell line exhibits increased sensitivity to the induction of cytogenetic aberrations by mitomycin C (MMC) relative to wild-type clone 707. It has been suggested that thymidine kinase-deficient cells may be highly mutagen-sensitive through an imbalance of nucleotide pools rendering excision repair error-prone. In this study clone 707 Friend leukaemia cells were compared with subclone 707BUF for sensitivity to the potentiating effect of caffeine on MMC-induced cytogenetic aberrations. The results indicate that although potentiation of mitomycin C-induced cytogenetic damage occurs in both clone 707 and in subclone 707BUF following caffeine treatment, the mutagen-sensitive thymidine kinase-deficient subclone 707BUF had enhanced potentiation by caffeine of MMC-induced cytogenetic damage relative to wild-type clone 707. It is suggested that caffeine may enhance mutagen sensitivity by inhibiting post-replication repair processes and may perhaps also indirectly reduce the effectiveness of the excision repair system by inhibiting the mutagen-induced G2-delay. Clone 707 wild-type cells in the presence of caffeine could then continue to repair DNA damage through an intact though less effective excision repair system, whilst the thymidine kinase-deficient subclone 707BUF would, in the presence of caffeine, be rendered highly mutagen sensitive, being only able to repair DNA damage through an error-prone excision repair process.

Evidence for indirect involvement of thymidine kinase in excision repair processes in mouse cell lines.

Wild-type cells and thymidine kinase-deficient clones from two mouse lymphoma cell lines, P388 and L5178Y, were compared for sensitivity to killing by the mutagens, ultraviolet irradiation (UV), ethyl methane sulfonate (EMS), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Two out of three thymidine kinase-deficient P388 clones showed significantly enhanced sensitivity to killing by all three mutagens. This increased sensitivity to killing was also reflected in increased mutagenesis by the three mutagens. In the L5178Y cell line, wild-type cells showed little difference to two thymidine kinase-deficient clones in terms of mutagen sensitivity. This indicates that thymidine kinase may be significant for DNA repair processes in P388 but not in L5178Y cells. Unscheduled DNA synthesis (UDS) experiments were carried out on P388 and L5178Y wild-type cells and wild-type Friend leukemia cells (which are mutagen-sensitive when deficient in thymidine kinase). The UDS experiments showed the L5178Y cells were low in excision repair abilities relative to the P388 cells and the Friend cell clone. This indicates that the increased mutagen sensitivity in thymidine kinase-deficient P388 and clone 707 Friend cells may be due to thymidine kinase playing an indirect role in DNA excision repair, a process which is of little significance in the L5178Y cell line.