Biosynthesis of basement membrane molecules by salivary adenoid cystic carcinoma cells: an immunofluorescence and confocal microscopic study

Abstract

The biosynthesis of basement membrane molecules and fibronectin was studied in vitro in the two different human cell systems (ACC2 and ACC3) established from adenoid cystic carcinomas (ACC) of the salivary gland using immunofluorescence and confocal microscopy. When cells were attached and spread on dishes, fine granular immunofluorescence for type IV collagen, laminin, heparan sulphate proteoglycan, entactin, and fibronectin first appeared diffusely in the cytoplasm, and then changed in aggregation of coarse granules in the perinuclear area. With formation of colonies, these signals were present in the extracellular space, initially in the basal aspect of attached cells and consequently in the lateral intercellular space. After the cells formed a confluent monolayer, extracellular signals started to decrease in inverse proportion to the reappearance of intracellular ones. The results indicate that the parenchymal cells of ACC synthesize these five extracellular matrix molecules, secrete them into the extracellular milieu and remodel the extracellular deposits. It is suggested that the characteristic stromal architecture of ACC, represented by stromal pseudocysts, results from their own secretion of the basement membrane molecules and fibronectin.

Promoter methylation and protein expression of the E-cadherin gene in the clinicopathologic assessment of adenoid cystic carcinoma

“…38 In vitro, ACC3 cell line also produces basement membrane molecules, including type IV collagen, laminin, heparan sulfate proteoglycan, and fibronectin. 6,28 It has been shown that E-cad methylation density of CpG sites increases when tumor cells are cultured on coated basement membrane molecules. 26 Excessive production of extracellular molecules in adenoid cystic carcinoma might thus be associated with a high incidence of E-cad gene methylation.…”

“…For the in vitro experiments, the ACC3 cell line, originally derived from a patient of adenoid cystic carcinoma arising from the parotid gland, was used. 28 Cultured tumor cells were maintained in RPMI-1640 medium (Gibco BRL, Rockville, MD, USA) supplemented with 10% fetal bovine serum (Gibco BRL), 1% L-glutamine, 50 mg/ml streptomycin, and 50 IU/ml penicillin, and incubated at 371C under a humidified atmosphere containing 5% CO 2 .…”

Podoplanin-mediated cell adhesion through extracellular matrix in oral squamous cell carcinoma

“…22 Salivary gland adenoid cystic carcinoma (ACC) cell systems (lines), ACC2 and ACC3, were established from ACCs of the palatal minor salivary gland and parotid gland, respectively, 23 and ACCM was subcloned from ACC2 cells as a clone with highly metastatic potential. 22 SCC cells were cultured in Dulbecco's modified Eagle medium (DMEM; Gibco, Invitrogen, Carlsbad, CA, USA) and ACC cells were cultivated in RPMI 1640 medium (Gibco), both of which contained 10% fetal bovine serum (Gibco), 50 mg/ml streptomycin, and 50 IU/ml penicillin (Gibco) (DMEMþ , RPMIþ ).…”

Knockdown of Sox4 expression by RNAi induces apoptosis in ACC3 cells

“…The molecular alterations that underlie the tumor development and progression are poorly understood. To determine if the high level of Sox4 gene expression plays a role in the pathogenesis of ACC, we used RNA silencing (RNAi) technology to inhibit Sox4 expression in the ACC-derived cell line ACC3 (Cheng et al, 1995) and assayed parameters of cell growth and apoptosis. We also identified relevant target genes that are differentially expressed in the Sox4 knockdown ACC3 using oligonucleotide microarray technology.…”