Origin and prognostic value of circulating KRAS mutations in lung cancer patients

Because of the current controversy on the origin and clinical value of circulating KRAS codon 12 mutations in lung cancer, we screened 180 patients using a combined restriction fragment-length polymorphism and polymerase chain reaction (RFLP-PCR) assay. We detected KRAS mutations in 9% plasma samples and 0% matched lymphocytes. Plasma KRAS mutations correlated significantly with poor prognosis. We validated the positive results in a second laboratory by DNA sequencing and found matching codon 12 sequences in blood and tumor in 78% evaluable cases. These results support the notion that circulating KRAS mutations originate from tumors and are prognostically relevant in lung cancer.

Regulation of Id1 expression by SRC: implications for targeting of the bone morphogenetic protein pathway in cancer

Deregulated activation of the Src tyrosine kinase and heightened Id1 expression are independent mediators of aggressive tumor biology. The present report implicates Src signaling as a critical regulator of Id1 gene expression. Microarray analyses showed that Id family genes were among the most highly down-regulated by incubation of A549 lung carcinoma cells with the small-molecule Src inhibitor AZD0530. Id1 transcript and protein levels were potently reduced in a dose-dependent manner concomitantly with the reduction of activated Src levels. These effects were conserved across a panel of lung, breast, prostate, and colon cancer cell lines and confirmed by the ability of PP2, Src siRNA, and Src-blocking peptides to suppress Id1 expression. PP2, AZD0530, and dominant-negative Src abrogated Id1 promoter activity, which was induced by constitutively active Src. The Src-responsive region of the Id1 promoter was mapped to a region 1,199 to 1,360 bps upstream of the translation start site and contained a Smad-binding element. Src was also required for bone morphogenetic protein-2 (BMP-2)-induced Id1 expression and promoter activity, was moderately activated by BMP-2, and complexed with Smad1/5. Conversely, Src inhibitors blocked Smad1/5 nuclear translocation and binding to the Src-responsive region of the Id1 promoter. Consistent with a role for Src and Id1 in cancer cell invasion, Src inhibitors and Id1 siRNA decreased cancer cell invasion, which was increased by Id1 overexpression. Taken together, these results reveal that Src positively interacts with the BMP-Smad-Id pathway and provide new ways for targeted inhibition of Id1.

The Src inhibitor AZD0530 blocks invasion and may act as a radiosensitizer in lung cancer cells

BACKGROUND: With the emergence of Src inhibitors in clinical trials, improved knowledge of the molecular responses of cancer cells to these agents is warranted. This will facilitate the development of tests to identify patients who may benefit from these agents, allow drug activity to be monitored and rationalize the combination of these agents with other treatment modalities. METHODS: This study evaluated the molecular and functional effects of Src inhibitor AZD0530 in human lung cancer cells, by Western blotting and reverse transcription-polymerase chain reaction, and by assays for cell viability, migration, and invasion. RESULTS: Src was activated in four of five cell lines tested and the level corresponded with the invasive potential and the histologic subtype. Clinically relevant, submicromolar concentrations of AZD0530 blocked Src and focal adhesion kinase, resulting in significant inhibition of cell migration and Matrigel invasion. Reactivation of STAT3 and up-regulation of JAK indicated a potential mechanism of resistance. AZD0530 gave a potent and sustained blockage of AKT and enhanced the sensitivity to irradiation. CONCLUSIONS: The results indicated that AZD0530, aside from being a potent inhibitor of tumor cell invasion which could translate to inhibition of disease progression in the clinic, may also lower resistance of lung cancer cells to pro-apoptotic signals.

Simultaneous quantitative detection of relevant biomarkers in breast cancer by quantitative real-time PCR

The assessment of ERa, PgR and HER2 status is routinely performed today to determine the endocrine responsiveness of breast cancer samples. Such determination is usually accomplished by means of immunohistochemistry and in case of HER2 amplification by means of fluorescent in situ hybridization (FISH). The analysis of these markers can be improved by simultaneous measurements using quantitative real-time PCR (Qrt-PCR). In this study we compared Qrt-PCR results for the assessment of mRNA levels of ERa, PgR, and the members of the human epidermal growth factor receptor family, HER1, HER2, HER3 and HER4. The results were obtained in two independent laboratories using two different methods, SYBR Green I and TaqMan probes, and different primers. By linear regression we demonstrated a good concordance for all six markers. The quantitative mRNA expression levels of ERa, PgR and HER2 also strongly correlated with the respective quantitative protein expression levels prospectively detected by EIA in both laboratories. In addition, HER2 mRNA expression levels correlated well with gene amplification detected by FISH in the same biopsies. Our results indicate that both Qrt-PCR methods were robust and sensitive tools for routine diagnostics and consistent with standard methodologies. The developed simultaneous assessment of several biomarkers is fast and labor effective and allows optimization of the clinical decision-making process in breast cancer tissue and/or core biopsies.

Das offene Selbstgespräch des Teampartners - Einfluss auf die wahrgenommene Kompetenz und die eigene Anstrengungsbereitschaft

Einleitung und Fragestellung In der Forschung zum Selbstgespräch im Sport dominiert die Frage nach dessen Wir-kung auf die Leistung. Der Einfluss des während der Leistungserbringung geäußerten Selbstgesprächs auf (un-)beteiligte Personen (z.B. Zuschauer, Gegenspieler oder Dop-pelpartner) wurde bislang kaum untersucht. Van Raalte, Brewer, Cornelius und Petitpas (2006) konnten immerhin zeigen, dass Zuschauer einen Tennisspieler als kompetenter einschätzen, wenn er in einem Laborexperiment mit positivem statt mit negativem Selbst-gespräch gezeigt wurde. In unserer Studie stehen Dyaden im Zentrum, in der eine Person eine andere beobach-tet, die mit sich selbst spricht. Untersucht wird, ob bei einer additiven Teamaufgabe sol-che Beobachtungen einen Effekt auf die Kompetenzeinschätzung und die Anstren-gungsbereitschaft haben. Es wird vermutet, dass die beobachtende Person dann eine höhere Anstrengungsbereitschaft (AV) zeigt, wenn sie einen Teampartner mit positivem Selbstgespräch (UV) beobachtet, da sie ihren Partner als kompetenter einschätzt und diese Einschätzung als Mediator wirkt. Methode Es wurde ein randomisierter Zweigruppen-Versuchsplan mit zwei Experimentalgruppen (je n=20) durchgeführt. Das Treatment bestand aus einem Video, das den (jeweils identi-schen) Teampartner bei der Bewältigung der zu absolvierenden visuomotorisch-koordinativen Testaufgabe zeigt (Test 2HAND des Wiener Testsystems). Das Video wur-de abhängig von der Gruppenzugehörigkeit mit negativen respektive positiven Selbstge-sprächsaussagen unterlegt. Die Mediatorvariable wahrgenommene Kompetenz wurde mittels eines Fragebogens erhoben, der in Anlehnung an Van Raalte et al. (2006) entwi-ckelt wurde. Das Vorgehen zur Erhebung der abhängigen Variable Anstrengungsbereit-schaft wurde von Ohlert (2009) übernommen: Die Probanden erhielten die Möglichkeit, sich auf die spätere Testphase vorzubereiten. Erfasst wurde dabei die Zeit, die sie für die Vorbereitung in Anspruch nahmen, und die Genauigkeit, mit der sie eine Reihe von Aufwärmaufgaben absolvierten. Ergebnisse Die Datenerhebung wurde soeben abgeschlossen. Die Ergebnisse der noch ausstehen-den regressionsanalytischen Auswertung werden auf der Tagung präsentiert. Literatur Ohlert, J. (2009). Teamleistung. Social Loafing in der Vorbereitung auf eine Gruppenaufgabe. Hamburg: Dr. Kovac. Van Raalte, J. L., Brewer, B. W., Cornelius, A. E. & Petitpas, A. J. (2006). Self-presentational effects of self-talk on perceptions of tennis players. Hellenic Journal of Psychology, 3, 134-149.