Hans-Joachim Terpe

Alpha 6 integrin distribution in human embryonic and adult tissues.

Alpha 6 integrin is an adhesion molecule that connects cells with extracellular matrix molecules of the laminin family. The laminin interaction seems to be essential for cell differentiation during embryogenesis and for the subsequent maintenance of tissue integrity in the adult. Alpha 6 integrin can also interact with laminin-independent cellular ligands and in this way plays a role in homing of leucocytes. Furthermore, in cancer biology α6 integrin has an important role in metastasis and as a possible new prognostic factor; exact knowledge of α6 integrin distribution in normal human tissues is therefore a crucial element. By immuno-histochemical methods we have screened α6 integrin expression of representative human tissues from the adult and the embryonic organism. All tested epithelia were α6 integrin positive, except for the endocrine cells of the pancreas and the adrenal glands. Heterogeneous staining was found on non-epithelial tissues. Strong staining was evident in peripheral nerves (Schwann cells), germ and Sertoli cells, endothelia, and smooth muscle cells of the myometrium. Weak staining was found in nerve cells of the stratum granulosum, the microglia, Kupffers cells and stromal cells of the ovary. All fibroblasts, striated muscle cells and astrocytes were negative. The tissue distribution of α6 integrin and the semi-quantitative estimation of their expression level should provide a better understanding of α6 integrin function under normal and phathological conditions, in particular in tumour progression.

Patterns of chromosomal aberrations in urinary bladder tumours and adjacent urothelium.

Bladder cancer is often characterized by recurrent and multifocal growth, and tumours are frequently accompanied by precancerous alterations of the surrounding urothelium. These findings have led to the hypothesis that cells from areas of genetically aberrant but morphologically non‐cancerous or even unremarkable mucosa may be the source of bladder carcinomas. Fluorescence in situ hybridization (FISH) was performed using ten probes targeting five different chromosomes that are known to be frequently altered in bladder cancer (centromere 1, 8, 9, 11, 17 and 1p36, 8p23, 9p21, 11q13, 17p13) on paraffin‐embedded tissue sections of 11 superficial bladder cancers. Copy number changes of the tumours were compared to those in the urothelium adjacent to the tumour. Eleven of 11 (100%) tumours and eight of 11 (73%) samples of adjacent urothelium showed copy number changes of at least one chromosome. The occurrence of similar patterns of chromosomal aberrations in the tumours and their associated urothelium supports the hypothesis of a clonal relationship. It is concluded that FISH analysis targeting five different chromosomes is more sensitive than conventional histology for distinguishing between neoplastic and normal cells of the urothelium. Copyright

Expression of cell adhesion molecules alpha-2, alpha-5 and alpha-6 integrin, E-cadherin, N-CAM and CD-44 in renal cell carcinomas : an immunohistochemical study

Renal cell carcinoma is known to metastasize early independent of tumour grade. Invasion of the renal vein plays an important role in the prognosis. Cell adhesion molecules have been investigated, including the expression of alpha-2, alpha-5, and alpha-6 integrin, E-cadherin, neural-cell adhesion molecule and CD-44 in 34 renal cell carcinomas, using the alkaline phosphataseantialkaline phosphatase technique. Our results indicate a differential expression of these cell adhesion molecules (alpha-2, alpha-5 and E-cadherin) depending on histological type and tumour grade.

Disseminated neuroblastomas under 1 year of age: cell biology and prognosis

Tumor specimens of 203 infants with neuroblastomas of different clinical stages — registered in successive multicenter clinical trials of the German Society of Pediatric Oncology — could be examined for N-myc amplification, chromosome 1-ploidy and — structure, CD44 std. expression (in tumor tissue, and also in patient‘s sera).Eightyseven (= 43%) of these infants had a non-localized, disseminated neuroblastoma, mainly involving sympathetic nerve tissue, lymphnodes, liver, skin, bone marrow and bones (46 patients were classified into the 4s group, 41 patients in the true 4 group).If the clinical classification between stage 4 and stage 4swas neglected, then 17 of these infants (= 20%) had N-myc amplification (4—64 copies) with 16 already dead. Seven of 9 examined patients with true stage 4 had chromosome 1p aberrations (with N-myc amplification in 5), and among the dead there were 2 with CD44 negative expression.In another series, serum CD44 std. was measured by ELISA, and the highest (significantly different) Kruskal-Wallis mean rank values (147.8) were found in infants (n = 6) with stage 4s compared to the low mean-rank-value of 71.9 in patients with stage 4 (n = 65). Stage 1—3 patients (n = 42) had values of 99.8—88.6.Thus, infants with disseminated neuroblastomas, showing non-diploidy, normal chromosome 1p structure, non-N-myc amplification and high CD44 std. expression in tumor tissue, and also high CD44 std. values in serum, will have the highest chance of survival due to tumor-non-progression.On the other hand, N-myc amplification in the tumor cells was found to be characteristic for stage 4s neuroblastoma patients with tumor progression (n=6). Therefore, 4s neuroblastoma-patients with N-myc amplified tumors should be aggressively treated like true stage 4 tumor patients!

Expression of cell adhesion molecules and tumor invasion in vitro by osteosarcoma cells

PROLIFERATION AND INVASIVENESS AS WELL AS EXPRESSION OF GAP JUNCTION CONNEXlNS OF TROPHOBLAST CELLS CHANGE WITH DIFFERENTIATION IN VIVO AND IN VITRO. R. Gr0mmer, B. Reu8, P. Hellmann, O. Traub, E. Winterhager Cell-cell communication via gap junctions plays an important role with regard to cell proliferation and differentiation. About 15 different gap junction connexins (cx) are known, but it is still not understood how this diversity is related to their function. We have investigated the pattern of connexin expression during trophoblast invasion and placental differentiation in the rat by immunohistochemistry and northern blot analysis. The invasive trophoblast cells of the ectoplacental cone express cx31, while during differentiation they switch to cx26 and cx43. This may point to an important role of those connexins in regulating the differentiation process of trophoblast lineages. To investigate the role of connexin expression for invasion and differentiation, we used two established cell lines, one corresponding to differentiated trophoblast cells (HRP; Hunt et al., Placenta 10: 161-177, 1989), and one rat choriocarcinoma cell line (RCHO; Faria and Soares, Endocrinol. 129: 2895-2906, 1991) which represents a malignant variant of the trophoblast. In parallel to our findings in the rat placenta, we could show that the undifferentiated RCHO cells are characterized by large amounts of cx31, while the differentiated HRP cells express cx43. With induction of differentiation by retinoic acid (RA; 10 -6 M) expression of cx31 is suppressed in RCHO cells, and in some cases, expression of cx43 was induced. Cell proliferation is greatly reduced and invasiveness is suppressed in a Matrigel invasion assay. In HRP cells application of RA did not alter connexin expression and only lead to a slight decrease of proliferation rate, but invasion was reduced compared to untreated cells. Thus the different connexin expression appears more likely to play a role in regulating the proliferation and differentiation than invasiveness of trophoblast cells.

CD44-Standard in human neuroblastoma: A new prognostic marker

CD44 VARIANT ISOPORMS ARE NOT ONLY EXPRESSED IN M E T A S T A S I Z I N G T U M O R C E L L S BUT ALSO IN EARLY E M B R Y O N I C D E V E L O P M E N T . C. Schw~irz ler , P. Ruiz , M. Kasper , M. V. Wiles and U. G0n the r t . CD44 isoforms have been shown to be expressed du r ing t u m o r p rogress ion in an ima l and h u m a n tumors . Invas ive t u m o r cells ut i l ize s imi la r m e c h a n i s m s as those used dur ing embryogenes i s a n d t i s sue remodel ing . Dur ing ear ly m a m m a l i a n d e v e l o p m e n t cell a d h e s i o n m o l e c u l e s a r e i n d i s p e n s a b l e for t h e embryo o rgan iza t ion . The t r a n s m e m b r a n e g lycopro te in CD44 w h i c h exis ts in mul t ip le isoforms is expressed dur ing t h e s e p r o c e s s e s . The var ious isoforms are gene ra t ed by a l te rna t ive spl icing of the p r e m R N A r e s u l t i n g in t he e n l a r g e m e n t of t h e ex t r ace l l u l a r p a r t of t he molecule. We p e r f o r m e d s e m i q u a n t i t a t i v e r e v e r s e t r a n s c r i p t a s e PCR followed by Sou the rn hybr id iza t ions wi th exon-specific probes to d e t e r m i n e t h e exon c o m p o s i t i o n s e x p r e s s e d as wel l as i m m u n o h i s t o c h e m i s t r y and whole m o u n t in s i tu hybr id iza t ions of m u r i n e , r a t a n d h u m a n e m b r y o s a t d i f f e r en t s t a g e s of deve lopment . The s t a n d a r d form of CD44 is widely expressed in a d u l t t i s sues and in embryos f rom l imb bud s t age onwards , whi le t he n u m e r o u s v a r i a n t i soforms exhib i t h ighly special ized p a t t e r n s of exp res s ion a l r eady in t h e egg cyl inder . The m o s t p r o m i n e n t t r a n s c r i p t s con ta in exons 6v-10v a n d 9v+10v. In l y m p h o h e m o p o i e s i s specific v a r i a n t i soforms also e m e r g e a t decisive d i f fe ren t ia t ion s tages. Al though specific l igands for t he ~ a r i a n t r eg ion s t i l l a w a i t i so la t ion , t h e h igh ly o r g a n i z e d ~xpress ion of CD44 v a r i a n t i soforms sugges t s a pivotal role in c e l l u l a r i n t e r a c t i o n s d u r i n g e a r l y d e v e l o p m e n t , p a t t e r n fo rmat ion and in hemopoiesis . Ref.: Ruiz, Sehw~rz l e r and Gi in the r t : C D 4 4 i s o f o r m s dur ing dif ferent ia t ion and development . BioEssays J an . 1995.

Expression of adhesion molecules related to tumourprogression in colorectal carcinoma: Grading, staging and follow up

CD44 VARIANT ISOPORMS ARE NOT ONLY EXPRESSED IN M E T A S T A S I Z I N G T U M O R C E L L S BUT ALSO IN EARLY E M B R Y O N I C D E V E L O P M E N T . C. Schw~irz ler , P. Ruiz , M. Kasper , M. V. Wiles and U. G0n the r t . CD44 isoforms have been shown to be expressed du r ing t u m o r p rogress ion in an ima l and h u m a n tumors . Invas ive t u m o r cells ut i l ize s imi la r m e c h a n i s m s as those used dur ing embryogenes i s a n d t i s sue remodel ing . Dur ing ear ly m a m m a l i a n d e v e l o p m e n t cell a d h e s i o n m o l e c u l e s a r e i n d i s p e n s a b l e for t h e embryo o rgan iza t ion . The t r a n s m e m b r a n e g lycopro te in CD44 w h i c h exis ts in mul t ip le isoforms is expressed dur ing t h e s e p r o c e s s e s . The var ious isoforms are gene ra t ed by a l te rna t ive spl icing of the p r e m R N A r e s u l t i n g in t he e n l a r g e m e n t of t h e ex t r ace l l u l a r p a r t of t he molecule. We p e r f o r m e d s e m i q u a n t i t a t i v e r e v e r s e t r a n s c r i p t a s e PCR followed by Sou the rn hybr id iza t ions wi th exon-specific probes to d e t e r m i n e t h e exon c o m p o s i t i o n s e x p r e s s e d as wel l as i m m u n o h i s t o c h e m i s t r y and whole m o u n t in s i tu hybr id iza t ions of m u r i n e , r a t a n d h u m a n e m b r y o s a t d i f f e r en t s t a g e s of deve lopment . The s t a n d a r d form of CD44 is widely expressed in a d u l t t i s sues and in embryos f rom l imb bud s t age onwards , whi le t he n u m e r o u s v a r i a n t i soforms exhib i t h ighly special ized p a t t e r n s of exp res s ion a l r eady in t h e egg cyl inder . The m o s t p r o m i n e n t t r a n s c r i p t s con ta in exons 6v-10v a n d 9v+10v. In l y m p h o h e m o p o i e s i s specific v a r i a n t i soforms also e m e r g e a t decisive d i f fe ren t ia t ion s tages. Al though specific l igands for t he ~ a r i a n t r eg ion s t i l l a w a i t i so la t ion , t h e h igh ly o r g a n i z e d ~xpress ion of CD44 v a r i a n t i soforms sugges t s a pivotal role in c e l l u l a r i n t e r a c t i o n s d u r i n g e a r l y d e v e l o p m e n t , p a t t e r n fo rmat ion and in hemopoiesis . Ref.: Ruiz, Sehw~rz l e r and Gi in the r t : C D 4 4 i s o f o r m s dur ing dif ferent ia t ion and development . BioEssays J an . 1995.