Definition of consolidation tumor ratio (B/A).

A: Maximal diameter of the consolidation part of the tumor was measured in CT cross section under lung window view. B: Maximal diameter of the consolidation part of the tumor was measured in CT cross section under mediastinal window.</p

Classifications of tumor component.

A. Pure ground glass opacity (GGO) B. Ground glass predominant C. Solid predominant D. Pure solid.</p

Disease free and overall survival for non small cell lung cancer patients for different C/T ratios.

A. Disease free survival for non small cell lung cancer patients with different C/T ratio (p <0.0001) B. Overall survival for non small cell lung cancer patients with different C/T ratio (p = 0.0043).</p

Multivariable analysis for disease free and overall survival (cox proportion hazard model).

Multivariable analysis for disease free and overall survival (cox proportion hazard model).</p

Exclusion algorithm.

Exclusion algorithm.</p

Correlation between CT tumor component and pathologic findings.

Correlation between CT tumor component and pathologic findings.</p

Descriptive data for non small cell lung cancer patients who received anatomic resection.

Descriptive data for non small cell lung cancer patients who received anatomic resection.</p

Tumor component vs. lymph node metastatic pattern in different tumor sizes.

Tumor component vs. lymph node metastatic pattern in different tumor sizes.</p

Comparison of IHC, FISH and RT-PCR Methods for Detection of <i>ALK</i> Rearrangements in 312 Non-Small Cell Lung Cancer Patients in Taiwan

<div><p>Background</p><p>Recently Echinoderm microtubule-associated protein-like 4- anaplastic lymphoma kinase (<i>EML4-ALK</i>) fusion gene has become an important biomarker for ALK tyrosine kinase inhibitor (crizotinib) treatment in NSCLC. However, the best detection method and the significance of <i>EML4-ALK</i> variant types remain uncertain.</p> <p>Methods</p><p>Reverse transcriptase-polymerase chain reaction (RT-PCR), fluorescence in Situ hybridization (FISH) and Immunohistochemical (IHC) stain were performed on tumor tissues of 312 NSCLC patients for detection of <i>ALK</i> rearrangements. Mutation analyses for <i>EGFR</i> and <i>KRAS</i> genes were also performed.</p> <p>Results</p><p>Thirteen of the 312 patients (4.17%) had <i>ALK</i> rearrangements detected by RT-PCR. If RT-PCR data was used as the gold standard, FISH tests had a low sensitivity (58.33%), but very good specificity (99.32%). IHC stain had better sensitivity (91.67%) than FISH, but lower specificity (79.52%), when the cut off was IHC2+. All of the 8 patients with high abundance of <i>EML4-ALK</i> positive cells in tumor tissues (assessed by the signal intensities of the RT-PCR product), were also have high expression of ALK protein (IHC3+), and positive for FISH, except one failed in FISH. Variants 3a+3b (4/5, 80%) of <i>EML4-ALK</i> fusion gene were more common to have high abundance of <i>EML4-ALK</i> positive cells in tumor tissues than variant 1 (1/3, 33.3%). Meta-analysis of the published data of 2273 NSCLC patients revealed that variant 3 (23/44, 52.3%) was the most common type in Chinese population, while variant 1 (28/37, 75.7%) was most common in Caucasian.</p> <p>Conclusions</p><p>Among the three detection methods, RT-PCR could detect not only the presence of <i>EML4-ALK</i> fusion gene and their variant types, but also the abundance of <i>EML4-ALK</i> positive cells in NSCLC tumor tissues. The latter two factors might affect the treatment response to anti-ALK inhibitor. Including RT-PCR as a diagnostic test for ALK inhibitor treatment in the prospective clinical trials is recommended.</p> </div

Quantitative Proteomics Reveals a Novel Role of Karyopherin Alpha 2 in Cell Migration through the Regulation of Vimentin–pErk Protein Complex Levels in Lung Cancer

Karyopherin alpha 2 (KPNA2) is overexpressed in various human cancers and is associated with cancer invasiveness and poor prognosis. Herein, to understand the essential role of KPNA2 protein complexes in cancer progression, we applied stable isotope labeling with amino acids in cell culture (SILAC)-based quantitative proteomic strategy combined with immunoprecipitation (IP) to investigate the differential KPNA2 protein complexes in lung adenocarcinoma cell lines with different invasiveness potentials. We found that 64 KPNA2-interaction proteins displayed a 2-fold difference in abundance between CL1-5 (high invasiveness) and CL1-0 (low invasiveness) cells. Pathway map analysis revealed that the formation of complexes containing KPNA2 and cytoskeleton-remodeling-related proteins, including actin, beta tubulin, tubulin heterodimers, vimentin, keratin 8, keratin 18, and plectin, was associated with cancer invasiveness. IP demonstrated that the levels of KPNA2–vimentin–pErk complexes were significantly higher in CL1-5 cells than in CL1-0 cells. The KPNA2–vimentin–pErk complex was also up-regulated in the advanced stage compared with the early-stage lung adenocarcinoma tissues. Importantly, the levels of pErk as well as cell migration ability were significantly reduced in KPNA2-knockdown cells; however, migration was restored by treatment with pErk phosphatase inhibitors. Collectively, our results demonstrate the usefulness of a SILAC-based proteomic strategy for identifying invasiveness-associated KPNA2 protein complexes and provide new insight into the KPNA2-mediated modulation of cell migration