Infiltration of tumour cells into cultures of isolated hepatocytes

1981 ◽  
Vol 47 (1) ◽  
pp. 385-397 ◽  
Author(s):  
E. Roos ◽  
I.V. Van de Pavert ◽  
O.P. Middelkoop

MB 6A lymphosarcoma and TA3 mammary carcinoma cells have previously been shown to infiltrate from liver blood vessels, where they had been arrested, into the liver parenchyma. The same tumour cells were previously added to cultures of isolated hepatocytes, 24 h after their isolation. Both tumour cell types adhered to both dorsal and lateral surfaces of hepatocytes. The lymphosarcoma cells rapidly infiltrated between the hepatocytes. They first extended pointed pseudopods between the liver cells, and when the tumour cell body intruded, they deeply invaginated the liver cells at an interhepatocyte boundary. The MB A6 cells accumulated between and under the hepatocytes, and after 24 h virtually all cells were contained within the cultures. TA3 cells also invaginated hepatocytes, not only at interhepatocyte boundaries, but all over the exposed surface. They did not extent pseudopods. The process was much slower than with MB 6A cells: After 24 h a few TA3 cells were completely encircled by hepatocytes. These observations indicate that the mechanism of infiltration is different from the 2 tumour cell types. Part of the TA3 cell did not invaginate the hepatocytes. Several of these cells spread on th hepatocyte surface, attaining a flattened shape. TA3 cells formed extensive tight junctions with the hepatocytes, sometimes sealing an intercellular lumen that resembled both a tumour acinus and a bile canaliculus. Also desmosomes were occasionally formed. The hepatocyte cultures appear to be a suitable model for studying the mechanism of liver infiltration, not only of tumour cells, but also leucocytes.

1982 ◽  
Vol 55 (1) ◽  
pp. 233-245
Author(s):  
E. Roos ◽  
de Pavert IV Van

The influence of tubulin-binding agents on the infiltration of murine MB6A lymphosarcoma and TA3 mammary carcinoma cells into primary rat hepatocyte cultures was studied. Colchicine, nocodazole and vinblastine reduced the number of infiltrating lymphosarcoma cells, probably by interfering with the adhesion of these cells to the exposed hepatocyte surface. However, the subsequent infiltration of cells that did adhere was not affected or even slightly stimulated. The reduced adhesion appears to be due to an effect on both the MB6A cells and the hepatocytes. In contrasts, adhesion of TA3 cells was not reduced and infiltration was markedly enhanced by these agents, due to an effect on the TA3 cells but probably not on the hepatocytes. These observations support previously described morphological evidence for differences between the infiltration mechanisms of the two tumour cell types. It is concluded that the system within the hepatocytes involved in adhesiveness of the exposed surface to MB6A cells is distinct from that mediating other types of adhesion. The tendency of TA3 cells to invaginate hepatocytes may be due to disturbances in tubulin-dependent processes.


2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A779-A779
Author(s):  
Michelle Williams ◽  
Jessica Christenson ◽  
Kathleen O’Neill ◽  
Sabrina Hafeez ◽  
Nicole Spoelstra ◽  
...  

BackgroundTo identify novel molecular mechanisms used by triple negative breast cancer (TNBC) to facilitate metastasis, we manipulated oncogenic epithelial-to-mesenchymal transition (EMT) by restoring the microRNA-200c (miR-200c), termed ‘the guardian of the epithelial phenotype.’ We identified several tumor cell catabolizing enzymes, including tryptophan 2,3-dioxygenase (TDO2) and heme oxygenase-1 (HO-1). The Richer lab has published that TDO2 promotes anchorage independent cell survival during TNBC metastasis via its catabolite kynurenine, which also induces CD8+ T cell death. Similarly, published studies have demonstrated that HO-1 supports BC anchorage independent survival. However, effects of the HO-1 catabolite bilirubin on the tumor microenvironment had not been studied. We postulated that TNBC utilize targetable catabolizing enzymes, like HO-1, to simultaneously support tumor cell survival and dampen the anti-tumor immune response.MethodsTo test our hypothesis in an immune competent mouse model, Met-1 mammary carcinoma cells from a late stage MMTV-PyMT tumor were engineered to inducibly express miR-200c. Tumor cell infiltrates were analyzed by immunohistochemistry (IHC), flow cytometry and multispectral fluorescence. RAW264.7 mouse macrophages were cultured with conditioned medium from carcinoma cells ± miR-200c or the HO-1 competitive inhibitor tin mesoporphyrin (SnMP). RAW264.7 macrophages were also treated with 0–20 µM bilirubin and macrophage polarization and efferocytic capacity, the ability to engulf dead tumor cells, were assessed using qRT-PCR and IncuCyte assays.ResultsMiR-200c restoration to Met-1 orthotopic tumors decreased growth by 45% and increased infiltration of CD11c+ dendritic cells and activation, determined by CD44 expression, of CD4+ and CD8+ T cells. While the number of F4/80+ macrophages was unchanged by miR-200c, the percent of M1 anti-tumor macrophages (F4/80+iNOS+/total cells) increased by >6-fold in miR-200c+tumors. RAW264.7 macrophages cultured with conditioned medium from miR-200c-restored mammary carcinoma cells had a 25–95% decrease in M2 pro-tumor genes (Arg1, Il4 and Il13) and a 15–55% increase in M1 genes (Nos2, Tnfa and Cxcl10). A similar decrease in M2 (30–50%) and increase M1 (35–160%) genes was seen in macrophages cultured with conditioned medium from SnMP treated mammary carcinoma cells. Conversely, bilirubin treatment alone enhanced M2 macrophage polarization and inhibited efferocytosis in a dose-dependent manner.ConclusionsUse of miR-200c to reverse EMT revealed that HO-1 promotes simultaneous TNBC cell survival and immune suppression. These studies are the first to show that tumor cell-HO-1 activity and subsequent bilirubin production may alter macrophage function in the tumor microenvironment. This finding could be clinically relevant since HO-1 inhibitors like SnMP are already FDA approved for treatment of other diseases.


2006 ◽  
Vol 24 (2) ◽  
pp. 151-157 ◽  
Author(s):  
Leif Viklund ◽  
Natalia Vorontsova ◽  
Tiina Henttinen ◽  
Markku Salmivirta

1991 ◽  
Vol 273 (1) ◽  
pp. 153-160 ◽  
Author(s):  
J F Coquil ◽  
B Berthon ◽  
N Chomiki ◽  
L Combettes ◽  
P Jourdon ◽  
...  

The monohydroxy bile acid taurolithocholate permeabilizes the endoplasmic reticulum to Ca2+ in rat liver cells. To assess whether this action on the endoplasmic reticulum was restricted to this tissue, the effects of bile acid were investigated in two cell types quite unrelated to rat hepatocyte, namely human platelets and neuronal NG108-15 cell line. The results showed that taurolithocholate (3-100 microM) had no effect on free cytosolic [Ca2+] in human platelets and NG108-15 cells. whereas it increased it from 180 to 520 nM in rat hepatocytes. In contrast, in cells permeabilized by saponin, taurolithocholate initiated a profound release of the stored Ca2+ from the internal Ca2+ pools in the three cell types. The bile acid released 90% of the Ca2+ pools, with rate constants of about 5 min-1 and half-maximal effects at 15-30 microM. The results also showed that, in contrast with liver cells, which displayed an influx of [14C]taurolithocholate of 2 nmol/min per mg, human platelets and the neuronal cell line appeared to be resistant to [14C]taurolithocholate uptake. The influx measured in these latter cells was about 100-fold lower than in rat liver cells. Taken together, these data suggest that human platelets and NG108-15 cells do not possess the transport system for concentrating monohydroxy bile acids into cells. However, they show that human platelets and neuronal NG108-15 possess, in common with liver cells, the intracellular system responsible for taurolithocholate-mediated Ca2+ release from internal stores.


Nature ◽  
1961 ◽  
Vol 192 (4799) ◽  
pp. 285-286 ◽  
Author(s):  
P. DE ◽  
R. CHATTERJEE ◽  
S. MITRA

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