William G. Chaney

Use of lectins as diagnostic and therapeutic tools for cancer

Within the past few years, lectins have become a well-established means for understanding varied aspects of cancer and metastasis. Evidence is now emerging that lectins are dynamic contributors to tumor cell recognition (surface markers), cell adhesion and localization, signal transduction across membranes, mitogenic stimulation, augmentation of host immune defense, cytotoxicity, and apoptosis. To advance understanding of these lectin-dependent processes, attempts are being made to discover new lectins that have one or more of these functions and to develop lectin- (or glycoconjugate-) based tools that could be used to home in on tumor cells. This review will summarize current research on the lectins and recent advances in the development of lectin-based diagnostic and therapeutic tools for cancer. Additionally, the future potential of lectin-based diagnosis and therapy is discussed.

Induction of N-acetylglucosaminyltransferase V by elevated expression of activated or proto- Ha-ras oncogenes

Viral infection of cultured cells with transforming viruses causes an increase in cell-surface N-linked β1-6 (GlcNAcβ1-6Man) branching of complex-type oligosaccharides. Similar observations have been made after transfection of cells with activated oncogenes, which is associated with an induction of tumorigenic and metastatic properties. In this study, the effects of transfection of both activated and proto-Ha-ras oncogenes into NIH3T3 cells were analyzed. The results showed that, in comparison with NIH3T3 cells, bothras transfectants have increased sensitivity to the cytotoxic action of L-PHA. An increase in β1-6 branching and an increased level of N-acetylglucosaminyltransferase V (GlcNAc-T V), the enzyme which initiates the β1-6 branching were also observed. The levels of GlcNAc-T I and β1-4 Gal-T remained unchanged in activated Ha-ras transfected NIH3T3 cells. These data suggest that a specific induction of GlcNAc-T V occurs after transfection with either the proto- or activated Ha-ras oncogenes, which is responsible for the increased β1-6 branching previously observed. (Mol Cell Biochem122: 85–92, 1993)

Localization of galectin-3 in normal and diseased pancreatic tissue

SummaryConclusionGalectin-3 is expressed in both human and hamster pancreatic tumors and tumor cell lines and this expression is increased over normal.BackgroundGalectin-3 is overexpressed in many gastrointestinal tumors. This study examined the expression of galectin-3 in human and hamster pancreatic tumors to determine if galectin-3 could be used as a marker for pancreatic cancer.MethodsMembranes were prepared from human and hamster pancreatic tumor cell lines. Galectin-3 was visualized by immunoblot analysis of separated membrane proteins using the monoclonal antibody (MAb) M3/38. Paraffin-embedded sections from normal, pancreatitis, and cancerous human pancreatic tissue and normal,N-nitrosobis (2-oxopropyl)amine (BOP)-treated hyperplastic, and cancerous hamster pancreatic tissues were processed immunohistochemically for galectin-3 using the MAb M3/38.ResultsGalectin-3 was heavily expressed in cytoplasmic and nuclear regions of 50% of normal human pancreatic tissue. Expression of galectin-3 in ductal cells in chronic pancreatitis and cancerous pancreatic tissue was increased over normal and was more uniform (>95% cells/duct stained). Normal hamster pancreatic ducts showed weak or no expression of galectin-3. Hyperplastic pancreatic ductal cells from BOP-treated hamsters heavily expressed galectin-3 (60–90% cells/duct stained). Galectin-3 expression in ductal cells in cancerous pancreatic lesions was increased to >95%. Galectin-3 was also detected in the pancreatic nerves in all human tissue specimens tested.

Tumor cell surface β1–6 branched oligosaccharides and lung metastasis

NIH3T3 cells transfected with an activated Ha-ras oncogene were treated with L-PHA, the leukoagglutinin from red kidney beans. Cell lines resistant to L-PHA-mediated cytotoxicity were isolated and found to contain reduced levels of L-PHA-binding oligosaccharides. The levels of N-acetylglucosaminyltransferase V, the enzyme responsible for the initiation of theβ1–6 branch, were reduced in L-PHA-resistant cells. Tumorigenicity in nude mice was unchanged by the change in oligosaccharide expression, but the ability to form lung tumors after intravenous injection was significantly reduced. These results demonstrate that the ability of NIH3T3 cells transfected with an activated Ha-ras oncogene to form lung tumors after intravenous injection into nude mice is reduced in all six L-PHA selected cell lines containing a reduction inβ1–6 branched Asn-linked oligosaccharides.

Human Prostatic Acid Phosphatase: Structure, Function and Regulation

Human prostatic acid phosphatase (PAcP) is a 100 kDa glycoprotein composed of two subunits. Recent advances demonstrate that cellular PAcP (cPAcP) functions as a protein tyrosine phosphatase by dephosphorylating ErbB-2/Neu/HER-2 at the phosphotyrosine residues in prostate cancer (PCa) cells, which results in reduced tumorigenicity. Further, the interaction of cPAcP and ErbB-2 regulates androgen sensitivity of PCa cells. Knockdown of cPAcP expression allows androgen-sensitive PCa cells to develop the castration-resistant phenotype, where cells proliferate under an androgen-reduced condition. Thus, cPAcP has a significant influence on PCa cell growth. Interestingly, promoter analysis suggests that PAcP expression can be regulated by NF-κB, via a novel binding sequence in an androgen-independent manner. Further understanding of PAcP function and regulation of expression will have a significant impact on understanding PCa progression and therapy.

Antiproliferative activity of novel imidazopyridine derivatives on castration-resistant human prostate cancer cells

Metastatic prostate cancer (mPCa) relapses after a short period of androgen deprivation therapy and becomes the castration-resistant prostate cancer (CR PCa); to which the treatment is limited. Hence, it is imperative to identify novel therapeutic agents towards this patient population. In the present study, antiproliferative activities of novel imidazopyridines were compared. Among three derivatives, PHE, AMD and AMN, examined, AMD showed the highest inhibitory activity on LNCaP C-81 cell proliferation, following dose- and time-dependent manner. Additionally, AMD exhibited significant antiproliferative effect against a panel of PCa cells, but not normal prostate epithelial cells. Further, when compared to AMD, its derivative DME showed higher inhibitory activities on PCa cell proliferation, clonogenic potential and in vitro tumorigenicity. The inhibitory activity was apparently in part due to the induction of apoptosis. Mechanistic studies indicate that AMD and DME treatments inhibited both AR and PI3K/Akt signaling. The results suggest that better understanding of inhibitory mechanisms of AMD and DME could help design novel therapeutic agents for improving the treatment of CR PCa.

Comparative studies on expression of tumor-associated antigens in human and induced pancreatic cancer in syrian hamsters

SummaryThe expression of blood-group-related antigens (BGRAs) in experimental primary pancreatic cancer induced byN-nitrosobis(2-oxopropyl)amine (BOP) treatment of Syrian hamsters and homologous subcutaneous transplants of this primary cancer in the cell line, PC-1, established from the primary cancer and intrapancreatic transplanted PC-1 cells were studied by histochemical and biochemical methods. Human primary pancreatic cancer; the human pancreatic cancer cell line, HPAF; and its subclones, CDU and CD18, also were studied on a comparative basis.Histochemical analysis of BGRAs demonstrated that A, B, H, Leb, Lex Ley, and T antigen were expressed both in vivo and in vitro in hamster and human materials in similar patterns. However, Lea, CA 19–9 and sialylated Tn antigens were not found in hamster-derived tissues. SDS-PAGE and Western blotting procedures using anti-A antigen revealed similar major bands in the membrane fractions of both human and hamster pancreatic cells between 97 and 200 kdalton. Among other human pancreatic cancerassociated antigens, TAG-72, CA 125, and 17–1A were detected immunohistochemically in the hamster tumors both in vivo and in vitro, in a pattern similar to that seen in human pancreatic cancer. Tumor antigen DU-PAN-2, associated with human pancreatic cancer, was found infrequently in hamster pancreatic cancer specimens. These results indicate that the experimental hamster pancreatic cancer model provides a unique tool for investigating antigenicity of pancreatic cancer, particularly in relation to diagnosis and therapy.

Modification of blood group a expression in human pancreatic tumor cell lines by inhibitors of N-glycan processing

SummaryPancreatic adenocarcinomas induced in Syrian hamsters byN-nitrosobis(2-oxopropyl)amine (BOP) treatment express blood group A (BGA) antigen, which was previously shown by this lab to be expressed on multiantennary asparagine (Asn)-linked glycans attached to membrane glycoproteins. To determine if a similar expression pattern was found in humans, three human pancreatic ductal adenocarcinoma cell lines (CD18, CD11, and Capan 1) from individuals of blood type A were analyzed and shown to express BGA antigen on membrane glycoproteins similar in molecular mass to those found in hamster tumor cells. The BGA antigen was located on Asn-linked oligosaccharides in all three human cell lines, as indicated by loss of activity after peptide:N-glycosidase F (PNGase F) treatment. Also, as shown previously in hamster pancreatic tumor cells, BGA expression at the surface of the human cell lines was blocked by growth of the cells in media containing deoxymannojirimycin (dMM), an inhibitor of mannosidase I. These results demonstrate that the BGA antigen is on Asn-linked glycans in human pancreatic adenocarcinoma cells and that these glycoproteins are processed similarly to the BGA glycoproteins in hamster pancreatic adenocarcinoma.

A novel adhesion factor produced by hamster pancreatic cancer cell line is effective on normal and carcinoma cell lines of different species.

SummaryAn adhesion factor, produced by the hamster pancreatic cancer cell line PC-1.0, was tested for its efficiency in promoting the in vitro adhesion of normal and tumor cells (pancreas, lung, kidney, colon, breast, skin, prostate, neuroblast, melanocyte) derived from human, monkey, bovine, hamster, and rat sources. Using a modification of the dimethylthyazol diphenyl tetrazolium (MTT) assay, the factor was found to induce adhesion in all cell lines in a dose-dependent manner. Although the effect was variously expressed, there was a statistically significant difference between the MTT absorbance of cells incubated in the presence or absence of the factor. Conditioned medium of each cell line tested showed significantly less adhesion effect than that produced by PC-1.0 cells. Because our previous study indicated that the adhesion factor produced by PC-1.0 cells differed from known growth factors and adhesion molecules including fibronectin, vibronectin, laminin, and collagen, it appears that PC-1.0 cells produce a novel adhesion factor that enhances adherence of normal and malignant cells of different species.