Morphological and functional differentiation of HSG cells: role of extracellular matrix and trpc 1. 2007

Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
Department of Restorative Dentistry and Periodontology, Dublin Dental School and Hospital, Trinity College, Dublin, Ireland. vag@fok.usn.hu

A human salivary intercalated duct cell line (HSG) is capable of morphological change to acinar-type cells, and of salivary amylase (AMY1) expression, by culturing on basement membrane extracts (BME). The aim of this study was to determine the critical conditions for functional and morphological differentiation of HSG cells and to establish if the processes are related. Cells were grown on BMEs that had different protein concentrations and growth factor content, and then examined with respect to morphology and AMY1 expression. To investigate the role of intracellular calcium in amylase expression, a pcDNA3.1-TRPC1alpha construct was used to overexpress htrp1alpha, which mediates the store-operated calcium entry in HSG cells. Expression of the AMY1, TRPC1alpha and beta genes was quantified by means of real time RT-PCR. Growth factor-reduced BME (12.8 mg/ml) induced multicellular acinar structures with lumen formation but without stimulation of either AMY1 or TRPC1. HSG cells cultured on higher concentration BME (17.5 or 16.4 mg/ml) formed reticular networks. AMY1 expression increased both on growth factor-reduced BME (17.5 mg/ml: 3.0-fold, P < 0.001) and on regular BME (16.4 mg/ml: 3.7-fold, P < 0.001) accompanied by a slight increase in expression of TRPC1alpha and TRPC1beta. Overexpression of htrp1alpha did not cause any significant changes in AMY expression, though it attenuated the BME (17.5 mg/ml)-induced AMY1 upregulation. Overall, the higher protein concentration BME favors amylase expression in HSG cells, whereas the lower concentration causes marked morphological changes.

UI MeSH Term Description Entries
D011494 Protein Kinases A family of enzymes that catalyze the conversion of ATP and a protein to ADP and a phosphoprotein. Protein Kinase,Kinase, Protein,Kinases, Protein
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002454 Cell Differentiation Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs. Differentiation, Cell,Cell Differentiations,Differentiations, Cell
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D004789 Enzyme Activation Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme. Activation, Enzyme,Activations, Enzyme,Enzyme Activations
D004815 Epidermal Growth Factor A 6-kDa polypeptide growth factor initially discovered in mouse submaxillary glands. Human epidermal growth factor was originally isolated from urine based on its ability to inhibit gastric secretion and called urogastrone. Epidermal growth factor exerts a wide variety of biological effects including the promotion of proliferation and differentiation of mesenchymal and EPITHELIAL CELLS. It is synthesized as a transmembrane protein which can be cleaved to release a soluble active form. EGF,Epidermal Growth Factor-Urogastrone,Urogastrone,Human Urinary Gastric Inhibitor,beta-Urogastrone,Growth Factor, Epidermal,Growth Factor-Urogastrone, Epidermal,beta Urogastrone
D005109 Extracellular Matrix A meshwork-like substance found within the extracellular space and in association with the basement membrane of the cell surface. It promotes cellular proliferation and provides a supporting structure to which cells or cell lysates in culture dishes adhere. Matrix, Extracellular,Extracellular Matrices,Matrices, Extracellular
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000681 Amylases A group of amylolytic enzymes that cleave starch, glycogen, and related alpha-1,4-glucans. (Stedman, 25th ed) EC 3.2.1.-. Diastase,Amylase
D001485 Basement Membrane A darkly stained mat-like EXTRACELLULAR MATRIX (ECM) that separates cell layers, such as EPITHELIUM from ENDOTHELIUM or a layer of CONNECTIVE TISSUE. The ECM layer that supports an overlying EPITHELIUM or ENDOTHELIUM is called basal lamina. Basement membrane (BM) can be formed by the fusion of either two adjacent basal laminae or a basal lamina with an adjacent reticular lamina of connective tissue. BM, composed mainly of TYPE IV COLLAGEN; glycoprotein LAMININ; and PROTEOGLYCAN, provides barriers as well as channels between interacting cell layers. Basal Lamina,Basement Lamina,Lamina Densa,Lamina Lucida,Lamina Reticularis,Basement Membranes,Densas, Lamina,Lamina, Basal,Lamina, Basement,Lucida, Lamina,Membrane, Basement,Membranes, Basement,Reticularis, Lamina

Related Publications

Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
October 2007, The British journal of dermatology,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
January 2011, Polish journal of veterinary sciences,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
January 2011, Journal of cellular biochemistry,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
January 1984, Progress in clinical and biological research,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
October 2005, Pflugers Archiv : European journal of physiology,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
June 1996, International journal of developmental neuroscience : the official journal of the International Society for Developmental Neuroscience,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
May 2005, Journal of cellular physiology,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
November 1994, Experimental cell research,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
January 2014, World journal of stem cells,
Janos Vag, and Elaine M Byrne, and Deirdre H Hughes, and Matthew Hoffman, and Indu Ambudkar, and Paula Maguire, and Brian C O'Connell
January 2009, Differentiation; research in biological diversity,
Copied contents to your clipboard!