MicroRNA Signature Predicts Survival and Relapse in Lung Cancer

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MicroRNA Signature Predicts Survival and Relapse in Lung Cancer Sung-Liang Yu, Hsuan-Yu Chen, Gee-Chen Chang, Chih-Yi Chen, Huei-Wen Chen, Sher Singh, Chiou-Ling Cheng, Chong-Jen Yu, Yung-Chie Lee, Han-Shiang Chen, Te-Jen Su, Ching-Cheng Chiang, Han-Ni Li, Qi-Sheng Hong, Hsin-Yuan Su, Chun-Chieh Chen, Wan-Jiun Chen, Chun-Chi Liu, Wing-Kai Chan, Wei J. Chen, Ker-Chau Li, Jeremy J.W. Chen, Pan-Chyr Yang Cancer Cell Volume 13, Issue 1, Pages 48-57 (January 2008) DOI: 10.1016/j.ccr.2007.12.008 Copyright © 2008 Elsevier Inc. Terms and Conditions

Figure 1 Kaplan-Meier Estimates of Overall Survival and Relapse-Free Survival of NSCLC Patients According to the MicroRNA Signature (A) Fifty-six patients in the training data set. (B) Fifty-six patients in the testing data set. (C) Sixty-two patients in the independent cohort. Cancer Cell 2008 13, 48-57DOI: (10.1016/j.ccr.2007.12.008) Copyright © 2008 Elsevier Inc. Terms and Conditions

Figure 2 MicroRNA Risk-Score Analysis of 112 NSCLC Patients (Upper panel) MicroRNA risk-score distribution. (Middle panel) Patients' survival status. (Bottom panel) Color-gram of microRNA expression profiles of NSCLC patients; rows represent high-risk and protective microRNAs, and columns represent patients. The blue dotted line represents the median microRNA signature cutoff dividing patients into low-risk and high-risk groups. Cancer Cell 2008 13, 48-57DOI: (10.1016/j.ccr.2007.12.008) Copyright © 2008 Elsevier Inc. Terms and Conditions

Figure 3 Kaplan-Meier Estimates of Overall Survival and Relapse-Free Survival According to the MicroRNA Signature in Subgroups of NSCLC Patients in the Combination of the Testing and Independent Data Sets (A) Stage I disease (n = 54). (B) Stage II disease (n = 24). (C) Stage III disease (n = 40). (D) Adenocarcinoma patients (n = 60). (E) Squamous cell carcinoma patients (n = 57). Cancer Cell 2008 13, 48-57DOI: (10.1016/j.ccr.2007.12.008) Copyright © 2008 Elsevier Inc. Terms and Conditions

Figure 4 MicroRNA Expression Alters the Invasiveness of Lung Cancer Cells In Vitro (A) The effect of microRNAs on cancer cell invasiveness. The low invasive lung cancer cell line (CL1-0) was transfected with high-risk microRNA precursors (hsa-miR-137, hsa-miR-182∗, or hsa-miR-372), and the high invasive cell line (CL1-5) was transfected with protective microRNA precursors (hsa-miR-221 and hsa-let-7a). Twenty-four hours after transfection, cells were seeded onto Transwell culture inserts and cultured for 18 hr. The invasive ability of transfected cells was measured by modified Boyden chamber assay. The number of invaded cells in cells transfected with negative control 1 precursor miRNA acts as microRNA control (miR-con), and relative invasiveness of each microRNA was normalized to miR-con. Each type of cell was assayed in quaternary. (B) Ectogenic microRNA expression quantified by real-time RT-PCR. Quantification of microRNA expression utilized total RNA isolated from cells transiently transfected with microRNA expression vector. The expression of microRNA was normalized to U6 RNA expression, and the microRNA transfectants were normalized to parental cell control (CL1-0 or CL1-5). Each type of cells was assayed in triplicate. (C) The effect of microRNAs on invasiveness evaluated by using ectogenic microRNA expression vectors. Invasion assay was performed as mentioned in (A). The number of invaded cells in cells transfected with pSilencer4.1-CMV puro negative control acts as microRNA control (miR-con). Each type of cells was assayed in quaternary. All data were processed by two-sided Student's t test and presented as mean ± SD. ∗ p < 0.05. Cancer Cell 2008 13, 48-57DOI: (10.1016/j.ccr.2007.12.008) Copyright © 2008 Elsevier Inc. Terms and Conditions