STAT3 silencing inhibits glioma single cell infiltration and tumor growth
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STAT3 silencing inhibits glioma single cell infiltration and tumor growth. / Priester, Maike; Copanaki, Ekaterini; Vafaizadeh, Vida; Hensel, Sandra; Bernreuther, Christian; Glatzel, Markus; Seifert, Volker; Groner, Bernd; Kögel, Donat; Weissenberger, Jakob.
In: NEURO-ONCOLOGY, Vol. 15, No. 7, 01.07.2013, p. 840-52.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - STAT3 silencing inhibits glioma single cell infiltration and tumor growth
AU - Priester, Maike
AU - Copanaki, Ekaterini
AU - Vafaizadeh, Vida
AU - Hensel, Sandra
AU - Bernreuther, Christian
AU - Glatzel, Markus
AU - Seifert, Volker
AU - Groner, Bernd
AU - Kögel, Donat
AU - Weissenberger, Jakob
PY - 2013/7/1
Y1 - 2013/7/1
N2 - BACKGROUND: Diffuse infiltration remains the fulcrum of glioblastoma's incurability, leading inevitably to recurrence. Therefore, uncovering the pathological mechanism is imperative. Because signal transducer and activator of transcription 3 (STAT3) correlates with glioma malignancy and predicts poor clinical outcome, we determined its role in glioma single cell infiltration and tumor growth.METHODS: STAT3 was silenced in Tu-2449 glioma cells via lentiviral gene transfer. Target gene expression was measured by real-time reverse transcription PCR, Western blotting, and immunohistochemistry. Microvilli were visualized by staining with wheat germ agglutinin. Migration and invasion were measured by Scratch and Matrigel chamber assays. Diffuse infiltration was studied in 350-μm-thick organotypic tissue cultures over 14 days using cells tagged with enhanced green fluorescent protein and live confocal laser scanning microscopy. Survival of tumor-bearing syngeneic, immunocompetent B6C3F1 mice was analyzed by Kaplan-Meier plots.RESULTS: STAT3 silencing reduced cell migration and invasion in vitro and stopped single cell infiltration ex vivo, while STAT3-expressing cells disseminated through the neuropil at ∼100 µm/day. STAT3 silencing reduced transcription of several tumor progression genes. Mice with intracranial STAT3 knockdown tumors had a significant (P< .0007) survival advantage over controls, yielding 27% long-term survival. STAT3 knockdown reduced podoplanin expression 50-fold and inhibited concurrent microvilli formation. STAT3 knockdown tumors exhibited a weaker podoplanin immunoreactivity compared with controls. Podoplanin staining was diffuse, preferentially at tumor margins, and absent in normal brain.CONCLUSIONS: Our results show compelling evidence that STAT3 is a key driver of diffuse infiltration and glioma growth and might therefore represent a promising target for an anti-invasive therapy.
AB - BACKGROUND: Diffuse infiltration remains the fulcrum of glioblastoma's incurability, leading inevitably to recurrence. Therefore, uncovering the pathological mechanism is imperative. Because signal transducer and activator of transcription 3 (STAT3) correlates with glioma malignancy and predicts poor clinical outcome, we determined its role in glioma single cell infiltration and tumor growth.METHODS: STAT3 was silenced in Tu-2449 glioma cells via lentiviral gene transfer. Target gene expression was measured by real-time reverse transcription PCR, Western blotting, and immunohistochemistry. Microvilli were visualized by staining with wheat germ agglutinin. Migration and invasion were measured by Scratch and Matrigel chamber assays. Diffuse infiltration was studied in 350-μm-thick organotypic tissue cultures over 14 days using cells tagged with enhanced green fluorescent protein and live confocal laser scanning microscopy. Survival of tumor-bearing syngeneic, immunocompetent B6C3F1 mice was analyzed by Kaplan-Meier plots.RESULTS: STAT3 silencing reduced cell migration and invasion in vitro and stopped single cell infiltration ex vivo, while STAT3-expressing cells disseminated through the neuropil at ∼100 µm/day. STAT3 silencing reduced transcription of several tumor progression genes. Mice with intracranial STAT3 knockdown tumors had a significant (P< .0007) survival advantage over controls, yielding 27% long-term survival. STAT3 knockdown reduced podoplanin expression 50-fold and inhibited concurrent microvilli formation. STAT3 knockdown tumors exhibited a weaker podoplanin immunoreactivity compared with controls. Podoplanin staining was diffuse, preferentially at tumor margins, and absent in normal brain.CONCLUSIONS: Our results show compelling evidence that STAT3 is a key driver of diffuse infiltration and glioma growth and might therefore represent a promising target for an anti-invasive therapy.
KW - Animals
KW - Apoptosis
KW - Blotting, Western
KW - Brain Neoplasms
KW - Cell Movement
KW - Cell Proliferation
KW - Gene Expression Regulation, Neoplastic
KW - Glioma
KW - Immunoenzyme Techniques
KW - Mice
KW - Microscopy, Confocal
KW - Organ Culture Techniques
KW - RNA, Messenger
KW - RNA, Small Interfering
KW - Real-Time Polymerase Chain Reaction
KW - Reverse Transcriptase Polymerase Chain Reaction
KW - STAT3 Transcription Factor
KW - Tumor Cells, Cultured
KW - Wheat Germ Agglutinins
U2 - 10.1093/neuonc/not025
DO - 10.1093/neuonc/not025
M3 - SCORING: Journal article
C2 - 23486688
VL - 15
SP - 840
EP - 852
JO - NEURO-ONCOLOGY
JF - NEURO-ONCOLOGY
SN - 1522-8517
IS - 7
ER -
