Overexpression of CARMA3 in Non-Small-Cell Lung Cancer Is Linked for Tumor Progression

We aimed to investigate the clinical significance of the expression of novel scaffold protein CARMA3 in non-small-cell lung cancer (NSCLC) and the biological function of CARMA3 in NSCLC cell lines. We observed moderate to high CARMA3 staining in 68.8% of 141 NSCLC specimens compared to corresponding normal tissues. The overexpression of CARMA3 was significantly correlated with TNM stage (P = 0.022) and tumor status (P = 0.013). CARMA3 upregulation also correlated with a shorter survival rate of patients of nodal status N0 (P = 0.042)as well as the expression of epidermal growth factor receptor (EGFR) (P = 0.009). In EGFR mutation positive cases, CARMA3 expression was much higher (87.5%) compared to non-mutation cases (66.1%). In addition, we observed that knockdown of CARMA3 inhibits tumor cell proliferation and invasion, and induces cell cycle arrest at the boundary between the G1 and S phase. We further demonstrated a direct link between CARMA3 and NF-κB activation. The change of biological behavior in CARMA3 knockdown cells may be NF-κB-related. Our findings demonstrated, for the first time, that CARMA3 was overexpressed in NSCLC and correlated with lung cancer progression, EGFR expression, and EGFR mutation. CARMA3 could serve as a potential companion drug target, along with NF-kB and EGFR in EGFR-mutant lung cancers

Thymosin beta 10 Prompted the VEGF-C Expression in Lung Cancer Cell

Background and objective Our previous study found that thymosin β10 overexpressed in lung cancer and positively correlated with differentiation, lymph node metastasis and stage of lung cancer. In this reasearch we aim to study the effects and mechanism of exogenous human recombinant Tβ10 on the expression of VEGF-C on non-small cell lung cancer. Methods After SPC, A549 and LK2 cells were treated with 100 ng/mL recombinant human Tβ10, the mRNA level of VEGF-C were detected by RT-PCR. The mean while the protein expression of VEGF-C, P-AKT and AKT were determined by Western blot assay. Results Exogenous recombinant human Tβ10 were significantly promote the expression levels of VEGF-C mRNA and protein while promoting the phosphorylation of AKT. Exogenous Tβ10 can promote the expression of VEGF-C mRNA and protein in lung cancer cell lines A549 and LK2 (P<0.05), and this effect can be inhibited by use AKT inhibitor LY294002 (P<0.05). Conclusion Tβ10 human recombinant proteins can promote the expression of VEGF-C by activating AKT phosphorylation in lung cancer cell lines

Distribution of CARMA3 status in NSCLC according to clinicopathological characteristics.

<p>Distribution of CARMA3 status in NSCLC according to clinicopathological characteristics.</p

Knockdown of CARMA3 expression in H1299 and A549 cells inhibited cell invasion in a Boyden chamber assay.

<p>(Upper) CARMA3 and Bcl10 siRNA treatment inhibited measurable cell invasion in both cell lines(200×). (Lower) The bar graph indicates the mean and difference in cell number among the photographs, and the standard derivation was calculated from the average of cells in the 10 fields (**, P<0.01).</p

Immunohistochemical staining of CARMA3 in lung cancer tissue sections.

<p>(A) Strong CARMA3 expression in lung adenocarcinoma(400×). (B) Weak CARMA3 expression in lung adenocarcinoma(400×). (C) Strong CARMA3 expression in lung squamous cell carcinoma(400×). (D) Weak CARMA3 expression in lung squamous cell carcinoma(400×). (E) Normal bronchial epithelium showing weak staining for CARMA3(400×). (F) Negative controls for CARMA3 staining with non-immune mouse IgG antibody(400×).</p

Examples of CAMRA3 and EGFR expression in NSCLC cases.

<p>Immunohistochemical staining for CARMA3 and EGFR in 2 representative NSCLC samples. One showed strong expression of CARMA3 (A) and strong EGFR expression (400×) (B). The other showed weak expression of CARMA3(400×) (C) and weak EGFR expression(400×) (D). (E,F) Correlation of CARMA3 and EGFR expression in NSCLC samples with EGFR mutation(400×).</p