Loss of heterozygosity of chromosome 9p21 is associated with the immortal phenotype of neoplastic human head and neck keratinocytes.

Human chromosomes 1,4,6, and 9 harbor genes which induce cellular senescence in vitro but a role for their inactivation in human tumors is not established. To investigate this we searched for loss of heterozygosity (LOH) on these chromosomes in keratinocyte cultures obtained from different stages of human squamous cell carcinoma progression. There was consistent LOH between markers D9S171 and D9S157 in 9 of 9 (100%) informative immortal cultures and in one line which entered crisis, but 0 of 7 informative senescent cultures showed LOH. These results suggest that inactivation of a gene at 9p21 is important but insufficient for human squamous cell carcinoma keratinocyte immortalization.

Evidence for the inactivation of multiple replicative lifespan genes in immortal human squamous cell carcinoma keratinocytes.

Human keratinocyte immortality is genetically recessive to the normal phenotype of limited replicative lifespan and appears to require the dysfunction of p53 and the cyclin D-Cdk inhibitor p16. In order to test for the inactivation of other candidate replicative lifespan genes in the immortal cells of human tumors, we developed a series of mortal and immortal keratinocyte cultures derived from neoplastic lesions of the head and neck which were amenable to molecular genetic analysis by the loss of heterozygosity (LOH) technique. The results indicate that keratinocyte immortalization in head and neck squamous cell carcinoma (SCC-HN) development involves the inactivation of at least two further pathways to senescence and four in all. Chromosomes 1, 4 and 7 carry genes representing immortality complementation groups C, B and D respectively and immortal keratinocytes showed LOH at either 4q32-q34 between D4S1554 and D4S171 (group B) or 7q31 (group D) but never 1q25 (group C). These results tentatively suggest that the genes responsible for the immortality complementation groups encode proteins on the same pathway to senescence. In addition, all of the immortal keratinocyte lines possessed high levels of telomerase activity and a suppressor of telomerase activity has been mapped to the short arm of chromosome 3p. Five out of eight lines showed LOH at 3p21.2-p21.3, a region which may carry a gene capable of suppressing SCC-HN telomerase. However, alternative mechanisms of telomerase reactivation were also suggested by our results. None of the above genetic alterations were seen in seven senescent neoplastic keratinocyte cultures. Other loci harbouring antiproliferative genes implicated in replicative lifespan showed few or no alterations and any alterations seen were additional to those described above.

Assignment of two human epidermal squamous cell carcinomas cell lines to more than one complementation group for the immortal phenotype.

Two human cell lines derived from squamous cell carcinomas (SCCs) of the epidermis, SCC-12 clone F and SCC-13 clone Y, were made to be independent of the Swiss 3T3 feeder layer to perform somatic-cell genetic experiments. We fused these SCC lines with normal human fibroblasts, and all resulting hybrids senesced after completing 12-17 population doublings, suggesting that in part, immortalization of the keratinocyte during SCC development results from the loss of gene function. We also tested whether these two SCC lines mapped to known complementation groups for immortality by fusing them with representatives of groups A (GM847), B (HeLa), and C (143B), but most of these hybrids were indistinguishable from those derived from homotypic crosses set up as immortal hybrid controls. As reported by others, fusions of cell lines from different complementation groups-143B (group C) x HeLa (group B) or GM847 (group A) x Hela (group B)--resulted in predominantly senescent hybrids. Our results confirmed and extended previous observations by others that the phenomenon of senescence is dominant to that of immortality, but they did not allow us to assign either of the SCC lines we studied to a complementation group for immortality.

Aversion substance(s) of the rat coagulating glands.

The aversive substance(s) present in adult male urine were not found in castrate rat urine. Removal of the coagulating glands also resulted in a loss of the aversion compounds. The aversion substances were restored to the urine after androgen treatment of the castrate rats.

Cellular immortality: a late event in the progression of human squamous cell carcinoma of the head and neck associated with p53 alteration and a high frequency of allele loss.

Many human tumors contain variant cells that, unlike their normal counterparts, possess indefinite proliferative potential in vitro. However, little is known of the relevance of these immortal cells to human carcinomas in vivo. To investigate immortality in a human tumor system, we established cultures from different stages of head and neck squamous carcinoma (SCC-HN). All the neoplastic cultures were transformed because they showed very low cornification in surface or suspension culture and were partially or completely resistant to suspension-induced death. Immortal variants were not detected in premalignant erythroplakia cultures, but their frequency increased with tumor progression, indicating that immortality is a late event in carcinogenesis. Some late-stage carcinomas still produced senescent cultures, but, significantly, all recurrent tumors were immortal. Immortal but not senescent carcinoma cultures were associated with p53 dysfunction and a high frequency of allele loss, indicative of tumor suppressor gene inactivation. These results show that there are at least two classes of human SCC-HN that are phenotypically and genotypically distinct and that the pathological stage of a given tumor is not necessarily indicative of the kind of cells it contains.

The Nithsdale schizophrenia surveys. X: Obstetric complications, family history and abnormal movements.

Obstetric histories of 54 schizophrenic patients and 114 siblings were obtained from their mothers and scored using the Obstetric Complications Scale. There were no statistically significant difference in the proportion of schizophrenic patients (35%) and siblings (29%) who had at least one definite obstetric complication. There was no evidence that schizophrenic patients with a history of obstetric complications were less likely to have a first-degree relative with a history of psychiatric illness leading to in-patient care. Schizophrenic patients with a history of obstetric complications were more likely to have drug-induced Parkinsonism. There was a trend for tardive dyskinesia to be more common in those schizophrenic patients with no obstetric complications but a family history of schizophrenia.