共查询到20条相似文献,搜索用时 31 毫秒
1.
Decreased connexin gene expression and loss of the capacity for either
homologous or heterologous intercellular communication has been associated
with neoplastic transformation. We tested the hypothesis that loss of gap
junctional intercellular communication (GJIC) correlates with tumorigenic
potential in the HeLa x skin fibroblast human hybrid cell system. Connexin
gene expression, gap junction function and tumorigenicity were determined
for the non-tumorigenic somatic hybrid cell line (CGL1) and a series of
UVC-induced tumorigenic cell lines derived from CGL1. CGL1 and the parental
skin fibroblasts express connexin43 (alpha1 gap junction gene) mRNA and
protein, form gap junctional plaques and have functional gap junctions.
UVC- irradiation of CGL1 cells produced a cell line (UV12) with an
aggressive tumorigenic phenotype, which lost connexin43 expression as well
as both homologous and heterologous GJIC and was in this respect similar to
HeLa cells. However, the phenotype of UV12 cells exhibited some instability
and revertants to a less aggressive tumorigenic phenotype were isolated.
These cells expressed connexin43 mRNA and protein, and demonstrated
homologous GJIC. Furthermore, cells reconstituted from a tumor derived from
this revertant cell line retained significant connexin43 expression and
homologous GJIC, although they exhibited an aggressive tumorigenic
phenotype. Thus, functional homologous GJIC cannot be dissociated from
tumorigenicity in this system. However, heterologous GJIC between these
same UVC-induced tumorigenic cell lines and normal human skin fibroblasts
was reduced, whereas the non-tumorigenic hybrid cells showed extensive
heterologous GJIC. In summary, re-acquisition of connexin43 expression and
homologous GJIC does not restore the non-tumorigenic phenotype in UVC-
induced tumorigenic HeLa skin fibroblast human hybrid cells. However,
reduction of heterologous GJIC does correlate with tumorigenicity in this
cell system.
相似文献
2.
G H Kalimi L L Hampton J E Trosko S S Thorgeirsson A C Huggett 《Molecular carcinogenesis》1992,5(4):301-310
We examined gap-junctional intercellular communication (GJIC) in a series of normal and v-raf-, v-myc-, and v-raf/v-myc-transduced rat liver epithelial (RLE) cell lines using the scrape loading-dye transfer and fluorescence-recovery-after-photobleaching (FRAP) assays. Whereas the normal RLE cell line, the control helper virus-transduced cell line, and the v-myc-transduced cell line all showed excellent GJIC, the v-raf-transduced cell lines displayed decreasing levels of GJIC associated with their increasing tumorigenicity. The v-raf/v-myc-transformed cell lines showed the lowest levels of GJIC and were also the most tumorigenic. Heterologous GJIC of these oncogene-transduced cell lines was also compared with that in the normal RLE cells. A modified FRAP assay, using fluorescent-microbead labelling to identify the oncogene-transduced cell from surrounding normal cells, was used to quantify the heterologous GJIC. The v-raf/v-myc-transformed RLE cells had no heterologous communication with the normal RLE cells, whereas v-raf- and v-myc-transduced cell lines maintained heterologous GJIC. Northern analysis showed that connexin 43 was the only gap-junction protein message expressed in these cell lines; connexin 32 and connexin 26 were not expressed. The levels of connexin 43 mRNA expression were relatively unchanged in all cell lines, suggesting that the reduction in GJIC was primarily at the posttranslational level. These findings suggest that reduction of homologous GJIC in v-raf- and v-raf/v-myc-transformed RLE cells is linked to their tumorigenic potential. Furthermore, the loss of heterologous GJIC, which we observed only in the v-raf/v-myc-transformed cells, might release such cells from the growth-regulating effects of surrounding normal cells, possibly contributing to their enhanced tumorigenic potential. 相似文献
3.
4.
The dysfunction of homologous and/or heterologous gap junctional intercellular communication (GJIC) has been implicated in tumorigenesis of many kinds of cells. Here we have characterized GJIC and the expression of connexins in six human lung carcinoma cell lines and normal lung fibroblasts (HLF). Compared with HLF, all the carcinoma cells showed reduced or little homologous GJIC. They expressed remarkably reduced connexin(Cx)43 mRNA and variable levels of Cx45 mRNA, but neither Cx43 nor Cx45 protein could be detected. However, using a preloading assay, transfer of calcein was observed between donor HLF cells and first order neighboring recipient tumor cells (recipient cells in 1000-fold excess). Transfer from tumor to HLF cells under the same conditions was not seen, although increasing the ratio of donor tumor cells to recipient HLF cells and plating the cells at low density did reveal weak transfer from tumor cells to HLF. Transfection of Cx43 into giant cell carcinoma PG cells increased homologous communication and eliminated the rectifying behavior of heterologous communication. This indicates that the apparent rectification of dye transfer between normal and tumor cells was a product of low rates of heterologous transfer linked to (i) rapid dilution of the dye to below detectable limits through a very well coupled cell population (tumor to HLF) and (ii) concentration of dye in immediate neighbors in a poorly coupled cell population (HLF to tumor cells). These results suggest that the coupling levels may need to exceed a certain threshold to allow propagation of signals over a sufficient distance to affect behavior of a cell population. We propose that the relative rates of heterologous and homologous coupling of cell populations and the 'pool size' of shared metabolites in tumor cells and the surrounding normal tissue are likely to be very important in the regulation of their growth. 相似文献
5.
Gap junctional intercellular communication and connexin43 expression in human ovarian surface epithelial cells and ovarian carcinomas in vivo and in vitro. 总被引:11,自引:0,他引:11
E A Hanna S Umhauer S L Roshong M P Piechocki M J Fernstrom J D Fanning R J Ruch 《Carcinogenesis》1999,20(7):1369-1373
Gap junctional intercellular communication (GJIC) and the expression of gap junction proteins (connexins) are frequently decreased in neoplastic cells and have been increased by cAMP and retinoids. GJIC and connexin expression were investigated in early passage normal human ovarian surface epithelial (HOSE) cells, human ovarian adenocarcinoma cell lines (CaOV-3, NIH:OVCAR-3, SK-OV-3 and SW626) and surgical specimens of human serous cystadenocarcinomas. We hypothesized that GJIC and connexin expression would be decreased in neoplastic cells and would be increased by cAMP and retinoic acid. Cultured HOSE cells exhibited extensive fluorescent dye-coupling and connexin43 (Cx43) expression; other connexins were not detected. The ovarian adenocarcinoma cell lines had little dye-coupling or connexin expression. Deletions and rearrangements of the Cx43 gene were not detected by Southern blotting in the carcinoma lines. N6, 2'-O-dibutyryladenosine 3',5'-cyclic monophosphate and all-trans-retinoic acid inhibited cell proliferation, but did not enhance GJIC or Cx43 expression. Surface epithelial cells of benign ovaries expressed Cx43, but this expression was barely detectable in ovarian serous cystadenocarcinomas. Thus, normal HOSE cells had extensive GJIC and Cx43 expression whereas ovarian carcinoma cells had less and cAMP and retinoic acid did not change these, although both agents inhibited cell growth. 相似文献
6.
A series of cells representing normal, non-tumorigenic cell lines, as well
as differentiating neoplastic and undifferentiated neoplastic rat tracheal
epithelial cell populations were evaluated for their ability to establish
homologous and/or heterologous cell-cell gap junction communication in
culture. Gap junction communication was evaluated by flow cytometric
quantitation of the transfer of the fluorescent dye calcein from a donor to
a recipient cell population via gap junctions. The data indicate that
normal primary cultures of rat tracheal epithelial cells, as well as
non-tumorigenic cell lines and squamous cell carcinomas cell populations,
retain the ability to establish both homologous and heterologous gap
junction communication. In all cases an average of >48% of recipient
cells had acquired calcein label during a 5-h interval of co-culture of
donor and recipient cells at confluent densities. Cells harvested directly
from squamous cell carcinoma tumors exhibited similar levels of cell-cell
communication. In contrast, cells giving rise to undifferentiated
carcinomas, as well as cells harvested from undifferentiated carcinomas,
exhibited very low levels or no homologous or heterologous cell-cell
communication. Cell populations exhibiting distinctly different
communication phenotypes were evaluated by Northern blot analysis for
expression of connexins (Cx 26, 32 and 43) and E-cadherin. Neither
communicating nor non-communicating cells expressed connexin 32. Those cell
populations, which established functional gap junctions, expressed
E-cadherin as well as connexin 26 and/or 43. In contrast, those cell
populations that lacked the ability to communicate universally lacked
expression of E-cadherin, and a quarter also lacked expression of
detectable levels of connexin.
相似文献
7.
8.
Kohji Yamakage Yasufumi Omori Colette Piccoli Hiroshi Yamasaki 《Molecular carcinogenesis》1998,23(2):121-128
Connexins are considered to be involved in cell growth control, on the basis of studies mainly with tumorigenic cells. To study the role of connexin genes in normal cell growth control, we established fibroblast cell lines from connexin 43 (Cx43)–deficient mice and characterized their growth. Embryonic fibroblasts from wild-type mice (Cx43+/+) and those with heterozygous (Cx43+/–) and homozygous (Cx43–/–) deficiencies of the Cx43 gene were cultured and passaged by a 3T3 protocol (every 3 d, 3 × 105 cells/60-mm dish). All cell lines showed a growth crisis during passages 6–15 and then started to grow well. All cell lines grew at similar rates under the 3T3 protocol, but Cx43-deficient (Cx43–/–) cell lines tended to grow faster when they were plated at 105 cells per dish. Cx43–/– cells did not express Cx43 and showed little gap-junctional intercellular communication (GJIC), confirming that Cx43 is the major connexin responsible for GJIC of these fibroblasts. While all Cx43+/+ and Cx43+/– cell lines expressed Cx43 protein, some of them showed very little GJIC. Those cell lines with high GJIC showed higher levels of the P2 form of Cx43 protein, and more Cx43 was localized in the plasma membrane than in cell lines with lower GJIC levels. We investigated effects of serum concentration on cell growth in these cell lines. Although different cell lines responded differentially to these agents, there was no clear relationship between Cx43 expression and cell growth stimulation by them. This suggests that Cx43 expression alone is not a strong regulator of mouse fibroblast growth. Mol. Carcinog. 23:121–128, 1998. © 1998 Wiley-Liss, Inc. 相似文献
9.
King TJ Fukushima LH Hieber AD Shimabukuro KA Sakr WA Bertram JS 《Carcinogenesis》2000,21(6):1097-1109
Loss of gap junctional intercellular communication (GJIC) has been linked to aberrant proliferation and an enhanced neoplastic phenotype. Many human tumors, including the cervical carcinoma line HeLa, have been reported to be deficient in expression of the gap junction protein connexin43 (Cx43) and GJIC. To determine if this is an early event in carcinogenesis, we utilized immunohistochemistry to screen a series of cervical biopsy samples and demonstrated a major reduction in Cx43 expression in dysplastic regions compared to normal epithelia. To determine whether this loss influences the neoplastic behavior of cervical carcinoma cells, we have constructed HeLa cell lines in which Cx43 expression can be induced in response to doxycycline. This approach allows for the discrimination of Cx43-mediated effects from those due to pre-existing clonal heterogeneity. Cx43 induction in these cells led to assembly of functional junctions but did not alter growth control in vitro as measured by logarithmic growth, saturation density or focus formation when in co-culture with growth-controlled fibroblasts. However, Cx43 induction decreased two indices of neoplasia: it reduced anchorage-independent growth and attenuated the growth rate of tumor xenografts. These results indicate that established HeLa cell lines are unresponsive to Cx43-mediated signals which are thought to mediate growth control of non-transformed cells, however, Cx43 expression can still reduce aspects of the neoplastic phenotype of these cells, indicating that loss of connexin signaling in dysplastic cells may contribute to their neoplastic progression. 相似文献
10.
连接蛋白基因Cx43抑制胶质瘤细胞增殖及其机理的初步探讨 总被引:9,自引:2,他引:7
目的 探讨连接蛋白基因Cx43对胶质瘤细胞增殖的抑制及其可能的机理。方法 将含Cx43cDNA的质粒以脂质体介导转染Cx43表达缺失的人和鼠的恶性胶质瘤细胞,通过Northem杂交、原位杂交及免疫组化染色检测Cx43mRNA及蛋白表达;MTT法测定细胞增殖率;核仁组成区嗜银蛋白染色检测细胞增殖活性;TUNEL法检测细胞凋亡;划痕标记荧光染料示踪技术检测细胞间隙连接通讯(GJIC);Western杂交及免疫组化染色检测bFGF、PDGF、EGFR、IGF-I和IGFBP3的表达。结果 转染Cx43基因的胶质瘤细胞增殖下降,GJIC恢复,同时伴有bFGF、PDGF、IGF-I和IGFBP3表达下降,而EGFR表达和细胞凋亡则无改变。结论 Cx43基因可能通过恢复GJIC功能及抑制某些重要生长因子的自分泌,实现对胶质瘤细胞增殖的抑制。 相似文献
11.
Differential effect of subcellular localization of communication impairing gap junction protein connexin43 on tumor cell growth in vivo 总被引:20,自引:0,他引:20
There is a large body of evidence suggesting the connexin gap junction proteins appear to act as tumor suppressors, and their tumor inhibitory effect is usually attributed to their main function of cell coupling through gap junctions. However, some cancer cells (e.g. the rat bladder carcinoma BC31 cell line) are cell-cell communication proficient. Using specific site-directed mutagenesis in the third membrane-spanning (3M) domain of connexin43 (Cx43), we abolished the intrinsic gap junction intercellular communication (GJIC) in BC31 cells either by closing the gap junctional channels or by disruption of the transport of connexin complexes to the lateral membrane. Clones of BC31 cells transfected with a dominant negative Cx43 mutant giving rise to gap junctional channels, permeable only for a small tracer (neurobiotin), displayed accelerated growth rate in vivo, showing the critical role of selective gap junctional permeability in the regulation of cell growth in vivo. The use of other dominant-negative mutants of Cx43 also suggested that the effect of impaired communication on the tumorigenicity of cancer cells depends on the subcellular location of connexin. Inhibition of intrinsic GJIC in BC31 cells by sequestering of Cx protein inside the cytoplasm, due to expression of dominant-negative transport-deficient Cx43 mutants, did not significantly enhance the growth of transfectants in nude mice, but occasionally slightly retarded it. In contrast, augmentation of GJIC in BC31 cells by forced expression of wild-type Cx43, or a communication-silent mutant, fully suppressed tumorigenicity of these cells. Overall, these results show that cell coupling is a strong, but not the sole, mechanism by which Cx suppresses growth of tumorigenic cells in vivo; a GJIC-independent activity of Cx proteins should be considered as another strong tumor-suppressive factor. 相似文献
12.
13.
14.
Gap junctional intercellular communication (GJIC) has been reportedto be markedly reduced in human mesothelloma tumour cell linescompared with primary mesothelial cells. Iminunofluorescencestainings have shown that the gap junction protein connexin43(Cx43) is expressed In both malignant and normal mesothelialcells. In this study the mRNA expression of Cx43 and three differentconnexlnsCx37, Cx40 and Cx45, which are highly expressedin lung tissuewas investigated in eight human mesotheliomacell lines, and in human primary mesothellal cells from severaldonors. The expression of the intercellular adhesion moleculesA-CAM (N-cadherln) and L-CAM (E-cadherin) was studied at theprotein level. No mRNA expression of Cx37, Cx40 or Cx45 in eithermesothelioma tumour cells or the primary mesothelial cells wasdetected. Cx43 was expressed at both the mRNA and the proteinlevel, in seven out of eight mesothelloma cell lines, as wellas in all the primary mesothellal cell cultures. The intercellularadhesion molecule A-CAM was expressed at the cellcellborders In six out of seven mesothelioma cell lines, as wellas in normal mesothellal cells. No expression of L-CAM was observedin these cells. The results suggest that Cx43 and A-CAM arethe major proteins in gap and adherens Junctions respectivelyin human mesothellal cells. Most mesothelioma tumour cell lineswith markedly reduced GJIC still express both Cx43 and A-CAM.Only one of our mesothelloma tumour cell lines severely deficientin GJIC lacks both the gap junction protein Cx43 and the celladhesion molecule A-CAM. 相似文献
15.
Numerous studies have demonstrated a correlation between dysregulation/loss of connexin expression or gap junction intercellular communication (GJIC) function and decreased growth control both in human tumors and tumor cell lines. Likewise, restoration of constitutive connexin expression/function is correlated with increased growth control/decreased tumorigenicity. Here, we show for the first time that inducible restoration of connexin43 (Cx43) expression and GJIC function in a human tumor line of mesenchymal origin (HT-1080, fibrosarcoma) resulted in a lowered neoplastic potential. Specifically, HT-1080 cells induced to express Cx43 demonstrated diminished foci formation when in co-culture with normal fibroblasts, decreased colony formation under anchorage-independent conditions, and reduced tumor growth when injected into immunodeficient mice. These results, obtained utilizing an inducible system that helps address issues of clonal heterogeneity, strongly implicate Cx43 as a tumor suppressor in human tissue of mesenchymal origin and GJIC as a regulatory mechanism for cellular growth control both in vitro and in vivo. This study also further supports the hypothesis that loss of Cx43/GJIC in human tumors may play an important role in the dysregulation of normal growth control. 相似文献
16.
Nelson K. S. Khoo Yuchun Zhang John F. Bechberger Shari L. Bond Kathleen Hum Peeyush K. Lala 《International journal of cancer. Journal international du cancer》1998,77(3):440-448
Poor gap junctional intercellular communication (GJIC) has been associated with uncontrolled cell growth and neoplasia. We have successfully propagated normal first trimester invasive extravillous trophoblast (EVT) cells, and have produced premalignant EVT lines after SV40 Tagtransformation: RSVT-2 is an uncloned line that is long-lived; RSVT2/C is a clonal line that is immortal. Both are hyperproliferative, hyperinvasive and variably refractory to the anti-proliferative and anti-invasive effects of transforming growth factor β (TGFβ). Possible changes in gap junctions during the transition of normal invasive EVT cells to the premalignant stage were examined by comparing expression of connexin proteins (by immunolabeling for Cx26, Cx32, Cx40, Cx43), and mRNA (by Northern blot with cDNA probes for Cx26, Cx32, Cx43), and functional GJIC (by dye transfer using the preloading method) in normal parental EVT cells and their SV40 Tag transformants. Results from immunofluorescence and Northern blot analysis revealed that, of the panel of connexins examined, only Cx43 was variably expressed in these cell lines in vitro. Expression of Cx43 protein and mRNA was abundant in normal EVT cell line HTR8, reduced in long-lived RSVT-2 cells and undetectable in immortalized RSVT2/C cells. GJIC, as measured by dye transfer between donor and recipient cells, was also similarly reduced in recipient RSVT-2 cells, and drastically reduced in RSVT2/C cells, irrespective of whether the dye donor was of the same cell type (homocellular coupling) or HTR8 cells (heterocellular coupling). Treatment with TGFβ reduced Cx43 mRNA expression as well as GJIC in normal EVT cells, but not in the SV40 Tag transformants. Our findings suggest that downregulation of connexins with the resultant impairment in GJIC is an early event in tumor progression, as observed in the premalignant SV40 Tag transformants. Int. J. Cancer77:440–448, 1998. © 1998 Wiley-Liss, Inc. 相似文献
17.
In this study, we investigated whether the tumor promoters, 12-O-
tetradecanoylphorbol-13-acetate (TPA), phenobarbital (PB), and 1,1-
bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT), inhibited gap junctional
intercellular communication (GJIC) in a cell-specific or connexin-specific
manner and whether protein kinase C was involved. To do this, we used
highly communicating WB-F344 rat liver epithelial cells, which express
connexin43 as their predominant gap junction protein, WB-aB1 cells, which
are a GJIC-incompetent mutant line of WB- F344 cells and that express
connexin43, WB-a/32-10 cells, which are a highly communicating derivative
of WB-aB1 cells generated by stable transduction with a connexin32
retroviral expression vector, and primary cultured rat hepatocytes, which
express conexin32 predominantly. Treatment of WB-F344 and WB-a/32-10 cells,
but not hepatocytes, with TPA inhibited GJIC (assayed by Lucifer Yellow dye
microinjection). This inhibition involved protein kinase C because (i)
inhibition was prevented by co-treatment of the cells with a specific
protein kinase C inhibitor, bis-indolylmaleimide, and (ii) treatment with
TPA for 24 h had no effect on dye-coupling in agreement with the
downregulation of protein kinase C. TPA also caused the internalization of
Cx43-containing gap junctions and the formation of a hyperphosphorylated
form of Cx43, Cx43-P3, in WB-F344 cells only, but TPA had no effect on
Cx32-containing gap junctions or protein mobility. In contrast, PB
inhibited GJIC only in hepatocytes and DDT inhibited GJIC in all three
types of cells; bis-indolylmaleimide did not block the effects of either
agent. These results indicate that the inhibitory actions of TPA and PB on
GJIC are cell-specific rather than connexin- specific and that TPA inhibits
connexin43 and connexin32-mediated GJIC through a protein kinase
C-dependent mechanism.
相似文献
18.
Expression and function of connexin in normal and transformed human keratinocytes in culture 总被引:4,自引:1,他引:3
Fitzgerald D.J.; Fusenig N.E.; Boukamp P.; Piccoli C.; Mesnil M.; Yamasaki H. 《Carcinogenesis》1994,15(9):1859-1865
We have studied the gap junctional intercellular communication(GJIC) of immortalized and tumourigenic human keratinocyte celllines and of a spontaneously immortalized non-tumourigenic anda highly differentiating keratinocyte cell line (HaCaT) as thecontrol. In homologous cultures, the GJIC capacity of five squamouscell carcinoma-derived cell lines was 127% that of theHaCaT cells. Ha-ras-transfected HaCaT cells with tumourigenicpotential and an SV40 DNA-immortalized cell line had markedlyreduced GJIC capacities. Northern analysis and immunohisto-chemistryshowed that connexin (Cx) 43 is the major gap junction proteinexpressed in the communicating cells. They do not express Cx26 or 32. The low or absent communication observed in certaincell lines was due in some to a lack of Cx 43 gene expression,but in others to aberrant localization of the gap junction protein.GJIC of these cell lines, as well as that of primary normalhuman epidermal keratinocytes, was susceptible to 12-O-tetra-decanoylphorbol-13-acetate-mediatedinhibition. Moreover, GJIC of HaCaT cells and their tumourigenicderivatives is Ca2+-dependent. These results, when comparedwith those previously obtained for mouse keratinocyte cell lines,reveal that GJIC of human keratinocytes was correlated to thedegree of differentiation and is controlled in a similar wayto that of murine keratinocytes. Aberrant GJIC seems to be acommon feature of human and murine skin carcinogenesis. 相似文献
19.
Vladimir A. Krutovskikh Hiroshi Yamasaki Hiroyuki Tsuda Makoto Asamoto 《Molecular carcinogenesis》1998,23(4):254-261
The tumor-suppressive property of the connexin gap-junction proteins was postulated from the fact that their function of cell coupling is impaired in most cancer cells. However, in conflict with this notion, certain cancer cells are able to communicate through gap junctions despite their malignancy. To explain this phenomenon, we studied by using a dominant-negative strategy the effect on tumorigenicity of loss of intrinsic gap-junction intercellular communication (GJIC) in the rat bladder carcinoma cell line BC31, which shows both expression of connexin 43 (Cx43) and intercellular communication. In cells transfected with a mutant Cx43 with seven residues deleted from the internal loop at positions 130–136 (Cx43Δ), transport of the resulting connexin protein to the plasma membrane occurred normally, but the GJIC of the cells was effectively abolished at the level of permeability of established gap junctions. Dominant-negative inhibition of GJIC by Cx43Δ accelerated growth of BC31 cells in nude mice. In contrast, when GJIC in BC31 cells was artificially enforced by transfection of wild-type Cx43, the cells lost the capacity to grow in vivo. Decreased phosphorylation of Cx43Δ suggested close interaction of the internal loop of connexin with its commonly phosphorylated domains in the C-terminal tail and involvement of this interaction in gap-junction permeability. Therefore, we conclude that the intrinsic GJIC observed in cancer cells should be considered a tumor-suppressor factor and that its level may influence malignant growth capacity. Mol. Carcinog. 23:254–261, 1998. © 1998 Wiley-Liss, Inc. 相似文献
20.
Diallyl disulfide (DADS) enhances gap-junctional intercellular communication by both direct and indirect mechanisms in rat liver cells 总被引:2,自引:0,他引:2
Huard C Druesne N Guyonnet D Thomas M Pagniez A Le Bon AM Martel P Chaumontet C 《Carcinogenesis》2004,25(1):91-98
Diallyl disulfide (DADS), a sulfur compound from garlic has been shown to exert many biological effects: induction of carcinogen detoxication, inhibition of tumor cell proliferation, etc. These effects are consistent with its anticarcinogenic properties in animal models and could account for garlic protective effects in humans. Our study demonstrates that DADS can improve gap-junctional intercellular communication (GJIC) in vitro. In rat liver epithelial cells (REL cells), using the dye transfer assay, we observe a time-dependent stimulation of GJIC by DADS at non-cytotoxic concentrations. In addition, incubation of cells with DADS for 1 h prevents the inhibition of GJIC induced by 3,5-di-tertio-butyl-4-hydroxytoluene (BHT). We have studied the direct effects of DADS on the regulation of GJIC, and especially on the expression and localization of the connexin expressed in these cells (Cx43): the enhancement of dye transfer (x1.6) by DADS from 1 to 50 micro M is associated with an increase (x1.3-1.8) in the amount of Cx43 protein (western blotting) with no alteration of its localization in the cell-cell contact regions of the plasma membrane (immunofluorescence analysis). We have also explored the possibility that DADS might act indirectly on GJIC. On one hand, DADS does not change the amount of E-cadherin, the adhesion molecule expressed in epithelial cells. On the other hand, it induces rapid inhibition of protein glycosylation. The data suggest that DADS could reduce local constraints imposed by glycoproteins, thus facilitating dye transfer. In conclusion, DADS can be included with other plant microconstituents, which have been demonstrated to improve GJIC. Its effect on REL cells can be explained by its ability to enhance the amount of Cx43 and also to diminish the level of glycosylated proteins. 相似文献