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TSC2 epigenetic defect in primary LAM cells. Evidence of an anchorage-independent survival.

Lesma E, Ancona S, Sirchia SM, Orpianesi E, Grande V, Colapietro P, Chiaramonte E, Di Giulio AM, Gorio A - J. Cell. Mol. Med. (2014)

Bottom Line: Moreover, LAM/TSC cells bear characteristics of stemness and secrete high amount of interleukin (IL)-6 and IL-8.Anti-EGF receptor antibodies and rapamycin affect proliferation and viability of non-adherent cells.In conclusion, the understanding of LAM/TSC cell features is important in the assessment of cell invasiveness in LAM and TSC and should provide a useful model to test therapeutic approaches aimed at controlling their migratory ability.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pharmacology, Dept. of Health Sciences, Università degli Studi di Milano, Milano, Italy.

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Lymphangioleiomyomatosis/tuberous sclerosis complex (LAM/TSC) cells secrete interleukins. (A) IL-6 (B) IL-8 and (C) IL-1α secretion were evaluated by Elisa in control LAM/TSC cells and after cycloheximide (CHX) overnight incubation. The effect of anti-EGF receptor (EGFR) antibody (EGFR Ab3), rapamycin (R1), methylprednisolone (MP) and repertaxin (Rep) after 48 hrs of incubation was evaluated. **P < 0.01 and ***P < 0.001 versus control LAM/TSC cells (anova followed by Bonferroni*s test). (D) IL-6 (E) IL-8 and (F) IL-1α release was evaluated after 5-azacytidine (5-aza) incubation. Error bars represent the SD for three experiments. Unpaired Student*s t-test p-values: *P < 0.05 versus control LAM/TSC cells.
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fig07: Lymphangioleiomyomatosis/tuberous sclerosis complex (LAM/TSC) cells secrete interleukins. (A) IL-6 (B) IL-8 and (C) IL-1α secretion were evaluated by Elisa in control LAM/TSC cells and after cycloheximide (CHX) overnight incubation. The effect of anti-EGF receptor (EGFR) antibody (EGFR Ab3), rapamycin (R1), methylprednisolone (MP) and repertaxin (Rep) after 48 hrs of incubation was evaluated. **P < 0.01 and ***P < 0.001 versus control LAM/TSC cells (anova followed by Bonferroni*s test). (D) IL-6 (E) IL-8 and (F) IL-1α release was evaluated after 5-azacytidine (5-aza) incubation. Error bars represent the SD for three experiments. Unpaired Student*s t-test p-values: *P < 0.05 versus control LAM/TSC cells.

Mentions: Tumour cells migrate from mouse primary tumours through a process of EMT, and this process is dependent on an inflammatory microenvironment [20]. Likewise, IL-6 promotes EMT in breast cancer cells, and SNAIL can induce IL-6 expression in keratinocytes [35,36]. Interleukin-8 and IL-6 production is regulated by PI3K and mitogen-activated protein kinase pathways, which are also involved in TSC cell deregulated functions [37–39]. Interleukin-1α, IL-6 and IL-8 were secreted by LAM/TSC cells as the inhibition of protein biosynthesis by incubation with cycloheximide prevented their secretion (Fig. 7A–C). The release of the interleukins was, however, not significantly affected by rapamycin or anti-EGFR antibody exposure. Interleukin-6, IL-8 and IL-1α secretion was inhibited by the incubation with 5-azacytidine leading to tuberin expression (Fig. 7D–F). As expected, methylprednisolone and repertaxin, an antagonist of IL-8 receptor, effectively reduced the release of interleukins [40] (Fig. 7A–C). Interleukin-1β was not detectable.


TSC2 epigenetic defect in primary LAM cells. Evidence of an anchorage-independent survival.

Lesma E, Ancona S, Sirchia SM, Orpianesi E, Grande V, Colapietro P, Chiaramonte E, Di Giulio AM, Gorio A - J. Cell. Mol. Med. (2014)

Lymphangioleiomyomatosis/tuberous sclerosis complex (LAM/TSC) cells secrete interleukins. (A) IL-6 (B) IL-8 and (C) IL-1α secretion were evaluated by Elisa in control LAM/TSC cells and after cycloheximide (CHX) overnight incubation. The effect of anti-EGF receptor (EGFR) antibody (EGFR Ab3), rapamycin (R1), methylprednisolone (MP) and repertaxin (Rep) after 48 hrs of incubation was evaluated. **P < 0.01 and ***P < 0.001 versus control LAM/TSC cells (anova followed by Bonferroni*s test). (D) IL-6 (E) IL-8 and (F) IL-1α release was evaluated after 5-azacytidine (5-aza) incubation. Error bars represent the SD for three experiments. Unpaired Student*s t-test p-values: *P < 0.05 versus control LAM/TSC cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4119383&req=5

fig07: Lymphangioleiomyomatosis/tuberous sclerosis complex (LAM/TSC) cells secrete interleukins. (A) IL-6 (B) IL-8 and (C) IL-1α secretion were evaluated by Elisa in control LAM/TSC cells and after cycloheximide (CHX) overnight incubation. The effect of anti-EGF receptor (EGFR) antibody (EGFR Ab3), rapamycin (R1), methylprednisolone (MP) and repertaxin (Rep) after 48 hrs of incubation was evaluated. **P < 0.01 and ***P < 0.001 versus control LAM/TSC cells (anova followed by Bonferroni*s test). (D) IL-6 (E) IL-8 and (F) IL-1α release was evaluated after 5-azacytidine (5-aza) incubation. Error bars represent the SD for three experiments. Unpaired Student*s t-test p-values: *P < 0.05 versus control LAM/TSC cells.
Mentions: Tumour cells migrate from mouse primary tumours through a process of EMT, and this process is dependent on an inflammatory microenvironment [20]. Likewise, IL-6 promotes EMT in breast cancer cells, and SNAIL can induce IL-6 expression in keratinocytes [35,36]. Interleukin-8 and IL-6 production is regulated by PI3K and mitogen-activated protein kinase pathways, which are also involved in TSC cell deregulated functions [37–39]. Interleukin-1α, IL-6 and IL-8 were secreted by LAM/TSC cells as the inhibition of protein biosynthesis by incubation with cycloheximide prevented their secretion (Fig. 7A–C). The release of the interleukins was, however, not significantly affected by rapamycin or anti-EGFR antibody exposure. Interleukin-6, IL-8 and IL-1α secretion was inhibited by the incubation with 5-azacytidine leading to tuberin expression (Fig. 7D–F). As expected, methylprednisolone and repertaxin, an antagonist of IL-8 receptor, effectively reduced the release of interleukins [40] (Fig. 7A–C). Interleukin-1β was not detectable.

Bottom Line: Moreover, LAM/TSC cells bear characteristics of stemness and secrete high amount of interleukin (IL)-6 and IL-8.Anti-EGF receptor antibodies and rapamycin affect proliferation and viability of non-adherent cells.In conclusion, the understanding of LAM/TSC cell features is important in the assessment of cell invasiveness in LAM and TSC and should provide a useful model to test therapeutic approaches aimed at controlling their migratory ability.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pharmacology, Dept. of Health Sciences, Università degli Studi di Milano, Milano, Italy.

Show MeSH
Related in: MedlinePlus