Salivary Gland Adenocarcinoma Cell Line Research Articles

ヒト顎下腺由来腺癌細胞株(HSG) レチノイン酸レセプターと レチノイン酸レスポンスエレメント における との結合

ヒト顎下腺由来腺癌細胞株(HSG) レチノイン酸レセプターと レチノイン酸レスポンスエレメント における との結合

Multidrug resistance-associated protein 7 expression is involved in cross-resistance to docetaxel in salivary gland adenocarcinoma cell lines.

The aim of the present study was to clarify whether ATP binding cassette transporters are refractory factors in head and neck cancers. For in vitro and in vivo chemotherapeutic studies, we used the following head and neck cancer cell lines: a mouse oral squamous cell carcinoma (SCC) cell line, Sq-1979; a human SCC cell line, SCCHA; a mouse salivary gland adenocarcinoma (SGA) cell line, NR-PG; and a human SGA cell line, HSY. We used a vinca alkaloid anticancer drug, vincristine (VCR), as a chemotherapeutic anticancer drug. To determine the cause of multidrug resistance, Western blot analysis, reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry of xenografted tumors in nude mice, drug efflux analysis, and drug efflux inhibitory assays were performed. VCR-treated cell lines, Sq-1979/VCR, SCCHA/VCR, NR-PG/VCR, and HSY/VCR, intensively expressed multidrug resistance (MDR) gene 1 mRNA and multidrug resistance associated protein (MRP) 1 mRNA. MRP7 mRNA and protein were expressed in NR-PG/VCR and HSY/VCR cells, but not in Sq-1979/VCR and SCCHA/VCR cells. In each cell clone of NR-PG/VCR and HSY/VCR, MRP7 mRNA was induced by VCR treatment, suggesting an acquired resistance to VCR in the context of MRP7 expression. In the in vivo chemotherapeutic nude mice model, VCR-treated xenografted SCCHA and HSY cells expressed MDR1 and MRP1. Moreover, MRP7 expression was immunohistochemically found in xenografted HSY cells of VCR-injected tumor-bearing mice, but not in SCCHA cells. Furthermore, doxorubicin accumulation was increased and drug cross-resistance to docetaxel decreased in HSY/VCR in the presence of a competitive MRP7 inhibitor, 17-beta-estradiol-(17-beta-D-glucuronide). These results indicate that MDR1 expression, MRP1 expression, and MRP7 expression are refractory factors in head and neck cancer chemotherapy and suggest that induction of MRP7 expression is involved in drug resistance to natural products, especially to docetaxel in SGA.

Refractory Factors in Head and Neck Cancer: ATP Binding Cassette Transporters Expressed in Head and Neck Cancer Cell Lines

The aim of the present study was to clarify whether ATP binding cassette transporters are refractory factors in head and neck cancer chemotherapy. For in vitro and in vivo chemotherapeutic studies, we employed a human salivary gland adenocarcinoma cell line (HSY) and a human oral squamous cell carcinoma cell line (SCCSK) with vincristine (VCR) at clinically equivalent doses. Western blot analysis, reverse transcription-polymerase chain reaction, in vivo evaluation in xenograft models inoculated with cultured carcinoma cell line and drug efflux analysis were performed. VCR-treated SCCSK and HSY cells, as well as xenografted SCCSK and HSY cells in tumor-bearing nude mice, were found to express MDR1/ABCB1 and MRP1/ABCC1. In addition to MDR1 and MRP1 mRNA, HSY/VCR and its cloned cells expressed MRP7/ABCC10 mRNA, but SCCSK/VCR did not express MRP7. Furthermore, drug resistance to VCR and docetaxel decreased in HSY/VCR in the presence of a competitive MRP7 inhibitor, 17-beta-estradiol-(17-beta-D-glucuronide). These results indicate that MDR1 and MRP1 expression are refractory factors in head and neck cancer chemotherapy and suggest that induction of MRP7 expression is involved in drug resistance in salivary gland adenocarcinomas.

Mechanism of Fas-mediated cell death and its enhancement by TNF-alpha in human salivary gland adenocarcinoma cell line HSG.

Fas-mediated cell death in a human salivary gland adenocarcinoma cell line (HSG) was induced by treatment of the cells with agonistic anti-Fas antibody (CH-11), and this cell death was enhanced by pretreatment with tumor necrosis factor alpha (TNF-alpha). The mode of cell death was apoptosis, because it was accompanied by caspase activation and the cleavage of poly(ADP-ribose) polymerase. The TNF-alpha treatment of the cells increased the expression of Fas, which was accompanied by the activation of nuclear factor kappaB (NFkappaB). These results suggest that the enhancement of the apoptosis caused by TNF-alpha resulted from increased sensitivity of the HSG cells to CH-11-mediated apoptosis due to induction of Fas protein by TNF-alpha via the activation of NFkappaB. In order to elucidate the apoptosis signaling pathway, we examined the effect of various caspase inhibitors on the apoptosis induced by CH-11. Fas-mediated apoptosis of HSG cells was slightly inhibited by the caspase-9 inhibitor although it was mainly inhibited by that for caspase-8. Based on this finding, we consider CH-11-induced apoptosis in HSG cells to be mainly mediated by the type I death signaling pathway that is caused by a caspase cascade initiated by the activation of caspase-8 at the death-inducing signaling complex (DISC).

Involvement of HSP90 in Anti-Fas-induced Apoptosis Signaling in the Human Salivary Gland Adenocarcinoma Cell Line HSG

It has been reported that heat shock protein (HSP) 90 is involved in the regulation of signaling cascades including those resulting in cell proliferation and apoptosis. However, little is known about how HSP90 affects apoptosis signaling.In this study, using a specific inhibitor of HSP90, geldanamycin (GDM), we investigated the relationship between HSP90 and anti-Fas antibody-induced apoptosis in HSG cells and concomitantly examined the apoptosis-inducing ability of GDM. We also sought to identify the target proteins of HSP90. When HSG cells were treated with GDM, a time-dependent increase in cell death was found. From the morphological features, the positive TdT-mediated dUTP nick end labeling (TUNEL) staining, and the cleavage of poly (ADP-ribose) polymerase (PARP), we concluded that the induced cell death was apoptotic. The pretreatment with GDM prior to that with anti-Fas antibody (CH-11) significantly increased the cell death as compared with that obtained with GDM or CH-11 alone. Further, prolonged incubation with GDM prominently enhanced the cell death. The induced cell death was also apoptotic. The transfection of HSG cells with recombinant HSP90α significantly inhibited the CH-11- and GDM-induced apoptosis. From the inhibition and overexpression experiments on HSP90 using GDM treatment and transfection with HSP90, respectively, we showed that HSP90 has an anti-apoptotic activity toward HSG cells. Immunoprecipitation with anti-human HSP90 antibody and subsequent Western blotting analysis of the precipitates detected bands for Caspase-8 and FADD-like ICE inhibitory protein (FLIP), which is known to regulate Fas cascade. Therefore, Caspase-8 and FLIP were shown to be target proteins of HSP90. These results suggest that HSP90 inhibits apoptosis by associating with Caspase-8 and FLIP and negatively regulating their functions.

Effect of salivary gland adenocarcinoma cell-derived α-N-acetylgalactosaminidase on the bioactivity of macrophage activating factor

The aim of this study was to clarify the effects of alpha-N-acetylgalactosaminidase (alpha-NaGalase) produced by human salivary gland adenocarcinoma (SGA) cells on the bioactivity of macrophage-activating factor (GcMAF). High exo-alpha-NaGalase activity was detected in the SGA cell line HSG. HSG alpha-NaGalase had both exo- and endo-enzyme activities, cleaving the Gal-GalNAc and GalNAc residues linked to Thr/Ser but not releasing the [NeuAc2-6]GalNac residue. Furthermore, GcMAF enzymatically prepared from the Gc protein enhanced the superoxide-generation capacity and phagocytic activity of monocytes/macrophages. However, GcMAF treated with purified alpha-NaGalase did not exhibit these effects. Thus, HSG possesses the capacity to produce larger quantities of alpha-NaGalase, which inactivates GcMAF produced from Gc protein, resulting in reduced phagocytic activity and superoxide-generation capacity of monocytes/macrophages. The present data strongly suggest that HSG alpha-NaGalase acts as an immunodeficiency factor in cancer patients.

ヒト顎下腺由来腺癌細胞株（ＨＳＧ）における抗Ｆａｓ抗体誘導アポトーシスシグナルへのＨＳＰ９０の関与

ヒト顎下腺由来腺癌細胞株（ＨＳＧ）における抗Ｆａｓ抗体誘導アポトーシスシグナルへのＨＳＰ９０の関与

Expression of cAMP Response Element Binding Protein (CREB)‐Binding Protein (CBP) and the Implication in Retinoic Acid‐Inducible Transcription Activation in Human Salivary Gland Adenocarcinoma Cell Line HSG

In the process of retinoic acid (RA) signaling, retinoic acid receptor interacts with a coactivator complex composed of various transcription cofactors such as CREB‐binding protein (CBP)/p300 and p160 family member proteins represented by steroid receptor coactivator‐1 (SRC‐1)/NCoA1 and p300/CBP cointegrator protein (p/CIP)/ACTR. In order to investigate the relationship of CBP to the RA signaling in a human salivary gland (HSG) adenocarcinoma cell line, we examined the expression of CBP in the cells. Immunoprecipitation and immunoblotting of the nuclear extract of HSG cells with anti‐human CBP antibody showed a specific 270‐kDa band, indicating the expression of CBP in HSG cells. The immunocytochemical analysis confirmed the nuclear localization of CBP. The transfection of HSG cells with a luciferase reporter plasmid harboring an RA‐response element at the 5′‐upstream region of the reporter gene increased RA‐dependent luciferase activity approximately 3‐fold. Co‐transfection with a CBP‐expression plasmid and the luciferase reporter gene enhanced the RA‐dependent transcription activation approximately 10‐fold. The immunoprecipitates obtained with anti‐CBP antibody exhibited a histone acetyl‐transferase (HAT) activity 2‐fold higher than that obtained with the control antibody, whereas the HAT activity of the immunoprecipitates with anti‐SRC‐1 and anti‐p/CIP, which were used as comparisons, were only a little increased. The RA treatment had no effect on the level of HAT activity except in the case of using the immunoprecipitate obtained with anti‐RARα, in which case it increased the activity. These findings indicate that CBP expressed in HSG cells mediates the RA‐inducible growth and differentiation‐regulating transcription activation in concert with the retinoic acid receptors.

Thermoradiotherapy combined with p53 gene therapy of human salivary gland adenocarcinoma cell line

We report on thermoradiotherapy combined with p53 gene therapy in the human salivary gland adenocarcinoma cell line HSG. HSG cells were successfully infected at a rate of 62.3% with MOIs of 20 of either AxCAip53 or AxCAiLacZ. The AxCAiLacZ did not inhibit the survival of the HSG cells. The survival fractions of AxCAip53-infected HSG cells were lower than those of the AxCAiLacZ-infected HSG cells, but there was no significant difference (p=0.30). AxCAip53 decreased the survival rates of thermotherapy (43 degrees C; p=0.084 and 44 degrees C; p=0.18), radiotherapy (6 Gy; p=0.20) and thermoradiotherapy (6 Gy plus 43 degrees C; p=0.24 and 6 Gy plus 44 degrees C; p=0.96), but there were no significant differences. In comparing the survival rates of thermoradiotherapy with those of thermotherapy and radiotherapy, thermoradiotherapy, regardless of the combination with p53 gene therapy, demonstrated actual survival rates lower than theoretical survival rates based on survival rates of thermotherapy multiplied by survival rates of radiotherapy. This result indicates that thermoradiotherapy is effective in the treatment of HSG cells. Thermoradiotherapy combined with p53 gene therapy was the most effective therapy among the combinations of therapies demonstrating that thermoradiotherapy combined with p53 gene therapy may be a useful tool in the treatment of HSG cells.

Thermochemotherapy of a human salivary adenocarcinoma cell line

We report on thermochemotherapy in a human salivary gland adenocarcinoma cell line. Hyperthermia reduced the survival rate to 50 and 20% by heating at 43 degrees C for 40 and 60 min, respectively, and is by itself useful in human salivary gland carcinoma treatment. Adriamycin, cisplatin, and mitomycin C can possible be used clinically, while bleomycin and 5-fluorouracil cannot, to treat this carcinoma. The optimal temperature was considered to be 41 degrees C in adriamycin, 42 degrees C in cisplatin, 37 degrees C in mitomycin C, and 42 degrees C in bleomycin in the thermochemotherapy. Thermochemotherapy is a useful tool in the treatment of human salivary gland carcinoma cells, but it is necessary to select the best anticancer drugs and the optimal temperature for optimal success using this treatment.

Inhibition of retinoic acid-inducible transcription by COUP-TFI in human salivary gland adenocarcinoma cell line HSG

Human salivary gland adenocarcinoma cells (HSG) express nuclear receptors, all-trans-retinoic acid (at-RA) receptors (RARs), and retinoid X/9-cis-retinoic acid (9-c-RA) receptors (RXRs). In order to investigate whether the endogenous RARs or RXRs of HSG cells can induce transcription activation, the thymidine kinase promoter (TK)-driven luciferase reporter gene containing the retinoic acid response element (RARE), of RARbeta, betaRARE2-TK-Luc, was transfected into HSG cells and ligand-dependent transcription activation was examined. Luciferase activity of cell lysate increased by the treatment with either at-RA or 9-c-RA. Co-transfection of RARalpha and (or) RXRalpha-expression plasmids with the reporter gene enhanced the luciferase activity, suggesting that endogenous RARs and RXRs work as ligand-dependent transfactors in HSG cells. Reverse transcriptase - polymerase chain reaction analysis revealed that HSG cells express chicken ovalbumin upstream promoter - transcription factor I (COUP-TFI). Co-transfection of COUP-TFI-expression plasmid suppressed the at-RA-induced transcription activation of the reporter gene. Similar results were shown using a chromatin-integrated reporter gene system, using a stably transfected beta-RARE2-TK-beta-galactosidase (beta-Gal) reporter gene. The at-RA-dependent increase in the beta-Gal expression was completely inhibited by COUP-TFI. The transfection of antisense oligonucleotide of COUP-TFI squelched the RA-dependent growth inhibition induced by RAR-RXR heterodimers. Conclusively, RARs and RXRs of HSG cells are functional and play roles as transactivators in at-RA-sensitive processes such as the proliferation or differentiation of cells. COUP-TFI very likely regulates these processes by repressing the functions of these transactivators.

Inhibition of retinoic acid-inducible transcription by COUP-TFI in human salivary gland adenocarcinoma cell line HSG

Inhibition of retinoic acid-inducible transcription by COUP-TFI in human salivary gland adenocarcinoma cell line HSG

Expression of retinoid X receptors and COUP-TFI in a human salivary gland adenocarcinoma cell line

The growth of the adenocarcinoma cell line derived from human salivary gland (HSG) is regulated by all-trans-retinoic acid (t-RA), which binds to its specific receptor, retinoic acid receptors (RARs), located in the nucleus, and thereby transactivates target genes. In this study, we examined the binding characteristics of the nuclear extract of HSG cells to the retinoic acid response element (RARE) compared with those of in vitro translated RAR alpha and retinoid X receptor alpha (RXR alpha), a heterodimeric partner of RAR alpha. Gel shift analysis using anti-RAR alpha and anti-RXR alpha antibodies revealed that the translated RAR alpha bound to RARE as a heterodimer with RXR alpha. In contrast, the binding of the nuclear extract of HSG cells to RARE showed a heterogeneous pattern, suggesting the existence of several species of RXRs as well as RARs in the nuclei of HSG cells. We therefore tried to clone these putative RXRs by the polymerase chain reaction using degenerated oligonucleotide primers conserved across the RXR family. The DNA sequencing of the recombinant clones revealed the expression of RXR alpha and RXR beta. In addition, chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI), which is also an RXR family member, was cloned. To evaluate the transcriptional activity of RARs and RXRs endogenously expressed in HSG cells, we performed a transient transfection analysis. When HSG cells were transfected with a luciferase reporter plasmid containing two repeats of either the RARE of the RAR beta gene or that of cellular retinol-binding protein II gene, positioned upstream of a thymidine kinase promoter fused to the luciferase sequence, a 2-3-fold induction of luciferase activity was observed in both cases. These results suggest that RARs and RXRs endogenously expressed in HSG cells were transcriptionally active in vivo. Thus, our findings showed that RXR alpha, RXR beta, and COUP-TFI are expressed in HSG cells and suggest that these molecules function as heterodimeric partners of RARs and (or) competitive repressors for RAREs and are involved in cellular responses mediated by retinoids.

Expression of retinoid X receptors and COUP-TFI in a human salivary gland adenocarcinoma cell line

The growth of the adenocarcinoma cell line derived from human salivary gland (HSG) is regulated by all-trans-retinoic acid (t-RA), which binds to its specific receptor, retinoic acid receptors (RARs), located in the nucleus, and thereby transactivates target genes. In this study, we examined the binding characteristics of the nuclear extract of HSG cells to the retinoic acid response element (RARE) compared with those of in vitro translated RAR alpha and retinoid X receptor alpha (RXR alpha ), a heterodimeric partner of RAR alpha . Gel shift analysis using anti-RAR alpha and anti-RXR alpha antibodies revealed that the translated RAR alpha bound to RARE as a heterodimer with RXR alpha . In contrast, the binding of the nuclear extract of HSG cells to RARE showed a heterogenous pattern, suggesting the existence of several species of RXRs as well as RARs in the nuclei of HSG cells. We therefore tried to clone these putative RXRs by the polymerase chain reaction using degenerated oligonucleotide primers conserved across the RXR family. The DNA sequencing of the recombinant clones revealed the expression of RXR alpha and RXR beta . In addition, chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI), which is also an RXR family member, was cloned. To evaluate the transcriptional activity of RARs and RXRs endogenously expressed in HSG cells, we performed a transient transfection analysis. When HSG cells were transfected with a luciferase reporter plasmid containing two repeats of either the RARE of the RAR beta gene or that of cellular retinol-binding protein II gene, positioned upstream of a thymidine kinase promoter fused to the luciferase sequence, a 2-3-fold induction of luciferase activity was observed in both cases. These results suggest that RARs and RXRs endogenously expressed in HSG cells were transcriptionally active in vivo. Thus, our findings showed that RXR alpha , RXR beta , and COUP-TFI are expressed in HSG cells and suggest that these molecules function as heterodimeric partners of RARs and (or) competitive repressors for RAREs and are involved in cellular responses mediated by retinoids. Key words: retinoid X receptor, retinoic acid receptor, retinoic acid response element, COUP-transcription factor (COUP-TF).

ヒト顎下腺由来腺癌細胞株の増殖と分化に及ぼすジブチリルｃＡＭＰの効果

ヒト顎下腺由来腺癌細胞株の増殖と分化に及ぼすジブチリルｃＡＭＰの効果

Retinoic acid receptor in subclone of human salivary gland adenocarcinoma cell line HSG and effect of retinoic acid on cellular growth

Retinoic acid (RA) binding has been detected in the nuclei of a subclone (CL-1) of human submandibular adenocarcinoma cell line HSG conditioned to grow in a serum-free defined medium. Competition assay confirmed the specificity of the RA binding. Scatchard analysis showed the binding molecule to have a high affinity and low capacity. From the analyses by gel-filtration and glycerol density gradient centrifugation, the nuclear binding molecule appears to be distinct from cellular RA binding protein (CRABP) in terms of molecular weight. Furthermore, immunoblotting analysis revealed a band ( M r 47 000) reactive with specific antibody to RA receptor (RAA)α in the gel containing the nuclear fraction of CL-1 cells. Northern blotting analysis with specific cDNA probes revealed the expression of RARα and RARγ in CL-1 cells. These results indicate that CL-1 cells express two types of RAR subtype, suggesting that these receptor molecules may mediate biological effects of RA. Treatment of CL-1 cells with RA resulted in an increase in the incorporation of [ 3H]thymidine into TCA-insoluble materials. The maximal increase was observed at 10 −6 M around 48 h. Previously, we demonstrated the autocrine growth of HSG cells mediated by epidermal growth factor (EGF) receptors and EGF-like molecules (Kurokawa et al. (1989) Cancer Res. 49, 5136–5142) and showed that RA had no significant effect on the secretion of the EGF-like molecule. RA induced an increase in [ 125I]EGF binding to CL-1 cells. The increase in the EGF binding was maximal at 24 h at 10 −6 M RA. RA also increased the amount of [ 3H]leucine-labeled EGF receptor dose-dependently. No significant change was observed in total protein synthesis of CL-1 cells by treatment with RA. These results suggest that RA stimulates the growth of CL-1 cells by increasing EGF receptor levels.

Effect of glucocorticoid on epidermal growth factor receptor in human salivary gland adenocarcinoma cell line HSG

Human salivary gland adenocarcinoma (HSG) cells treated with 10 −6 M triamcinolone acetonide for 48 h exhibited a 1.7- to 2.0-fold increase in [ 125I]human epidermal growth factor (hEGF) binding capacity as compared with untreated HSG cells. Scatchard analysis of [ 125I]EGF binding data revealed that the number of binding sites was 83 700 (± 29 200) receptors / cell in untreated cells and 160 500 (± 35 500) receptors / cell in treated cells. No substantial change in receptor affinity was detected. The dissociation constant of the EGF receptor was 0.78 (± 0.26) · 10 −9 M for untreated cells, whereas it was 0.93 (± 0.31) · 10 −9 M for treated cells. The triamcinolone acetonide-induced increase in [ 125I]EGF binding capacity was dose-dependent between 10 −9 and 10 −6 M, and maximal binding was observed at 10 −6 M. EGF receptors on HSG cells were affinity-labeled with [ 125I]EGF by use of the cross-linking reagent disuccinimidyl suberate (DSS). The cross-linked [ 125I]EGF was 3–4% of the total [ 125I]EGF bound to HSG cells. The affinity-labeled EGF receptor was detected as a specific 170 kDa band in the autoradiograph after SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Densitometric analysis revealed that triamcinolone acetonide amplified the intensity of this band 2.0-fold over that of the band of untreated cells. EGF receptor synthesis was also measured by immunoprecipitation of [ 3H]leucine-labeled EGF receptor protein with anti-hEGF receptor monoclonal antibody. Receptor synthesis was increased 1.7- to 1.8-fold when HSG cells were treated with 10 −8–10 −6 M triamcinolone acetonide for 48 h. When the immunoprecipitated, [ 35S]methionine-pulse-labeled EGF receptor was analyzed by SDS-PAGE and fluorography, the newly synthesized EGF receptor was detected at the position of 170 kDa; and treatment of HSG cells with triamcinolone acetonide resulted in a 2.0-fold amplification of this 170 kDa band. There was no significant difference in turnover rate of EGF receptor between treated and untreated HSG cells. These results demonstrate that the triamcinolone acetonide-induced increase in [ 125I]EGF binding capacity is due to the increased synthesis of EGF receptor protein in HSG cells.

Nonactivated and activated glucocorticoid receptor complexes from human salivary gland adenocarcinoma cell line

[ 3H]Triamcinolone acetonide glucocorticoid receptor complexes from human salivary gland adenocarcinoma cells (HSG cells) were shown to be activated with an accompanying decrease in molecular weight in intact cells, as analyzed by gel filtration, DEAE chromatography, the mini-column method and glycerol gradient centrifugation. Glucocorticoid receptor complexes consist of steroid-binding protein (or glucocorticoid receptor) and non-steroid-binding factors such as the heat-shock protein of molecular weight 90 000. To determine whether the steroid-binding protein decreases in molecular weight upon activation, affinity labeling of glucocorticoid receptor in intact cells by incubation with [ 3H]dexamethasone 21-mesylate, which forms a covalent complex with glucocorticoid receptor, was performed. Analysis by gel filtration and a mini-column method indicated that [ 3H]dexamethasone 21-mesylate-labeled receptor complexes can be activated under culture conditions at 37°C. SDS-polyacrylamide gel electrophoresis of [ 3H]dexamethasone 21-mesylate-labeled steroid-binding protein resolved only one specific 92 kDa form. Furthermore, only one specific band at 92 kDa was detected in the nuclear fraction which was extracted from the cells incubated at 37°C. These results suggest that there is no change in the molecular weight of steroid-binding protein of HSG cell glucocorticoid receptor complexes upon activation and that the molecular weight of nuclear-binding receptor does not change, although the molecular weight of activated glucocorticoid receptor complexes does decrease. Triamcinolone acetonide induced an inhibitory effect on DNA synthesis in HSG cells. Dexamethasone 21-mesylate exerted no such effect and blocked the action of triamcinolone acetonide on DNA synthesis. These results suggests that dexamethasone 21-mesylate acts as antagonist of glucocorticoid in HSG cells. The fact that dexamethasone 21-mesylate-labeled receptor complexes could be activated and could bind to DNA or nuclei aas well as triamcinolone acetonide-labeled complexes suggests that dexamethasone 21-mesylate-labeled complexes can not induce specific gene expression after their binding to DNA.

Enhancement of cis-diamminedichloroplatinum and mitomycin C-induced sister chromatid exchanges by dibutyryl cyclic AMP in a human salivary gland adenocarcinoma cell line in culture.

A human salivary gland adenocarcinoma cell line was cultivated in the presence of dibutyryl cyclic AMP (dB-cAMP), cis-diammine dichloroplatinum (cisplatin) or mitomycin C (MMC) only, or of the combination of dB-cAMP and each of the antineoplastic drugs. Then the treated cells were examined for the induction of sister chromatid exchanges (SCEs), colony-forming efficiency (CFE) in semisolid agar medium and cell survival as measured by counting for viable cells. It has been found that the frequency of SCE induction is significantly increased by treatment of cells with cisplatin or MMC under the presence of dB-cAMP, when compared to that in the cells treated with cisplatin or MMC only. Moreover, marked reduction in CFE and cell survival was found to occur in the cells treated with the combination of dB-cAMP and cisplatin or MMC. These finding indicate that treatment of a human salivary gland adenocarcinoma cell line with dB-cAMP results in modification of SCE induction, CFE and cell survival in the cells exposed to cisplatin or MMC.

Glucocorticoid regulates secretion of epidermal growth factor in the human salivary gland adenocarcinoma cell line.

The treatment of a human submandibular gland adenocarcinoma cell line (HSG cell line) for 48 h with triamcinolone acetonide (TA; 1-100 nmol/l) reduced the secretion of epidermal growth factor (EGF) in a closely related manner to a maximum of 66%. The reduction in the level of EGF secreted resulted in the suppression of DNA synthesis in the HSG cells to a similar extent. When the cells were incubated with TA and exogenous human EGF (hEGF), DNA synthesis was 1.7-fold higher than that without added hEGF. The removal of EGF by the addition of hEGF antibody reduced DNA synthesis in HSG cell cultures to the same extent as did TA. These results suggest that the growth inhibition of HSG cells by TA is due to the reduction in the amount of EGF secreted.