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Expression levels of  miR-146b-5p  and  -3p  microRNAs in human non-small cell lung cancer (NSCLC) are associated with recurrence of the disease after surgery. To understand this, the effect of  miR-146b  overexpression was studied in A549 human lung cancer cells.    Methods  A549 cells, engineered with lentiviruses to overexpress the human  pre-miR-146b  precursor microRNA, were examined for proliferation, colony formation on plastic surface and in soft agar, migration and invasiveness in cell culture and in vivo in mice, chemosensitivity to cisplatin and doxorubicin, and global gene expression.  miR-146b  expressions were assessed in microdissected stroma and epithelia of human NSCLC tumors. Association of  miR-146b-5p  and  -3p  expression in early stage NSCLC with recurrence was analyzed.    Principal Findings  A549  pre-miR-146b -overexpressors had 3–8-fold higher levels of both  miR-146b  microRNAs than control cells. Overexpression did not alter cellular proliferation, chemosensitivity, migration, or invasiveness; affected only 0.3% of the mRNA transcriptome; and, reduced the ability to form colonies in vitro by 25%. In human NSCLC tumors, expression of both  miR-146b  microRNAs was 7–10-fold higher in stroma than in cancerous epithelia, and higher  miR-146b-5p  but lower  -3 p levels were predictive of recurrence.    Conclusions  Only a minimal effect of  pre-miR-146b  overexpression on the malignant phenotype was seen in A549 cells. This could be because of opposing effects of  miR-146b-5p  and  -3p  overexpression as suggested by the conflicting recurrence-predictive values of the two microRNAs, or because  miR-146b  expression changes in non-cancerous stroma and not cancerous epithelia of tumors are responsible for the prognostic value of  miR-146b .

Overexpression of the Lung Cancer-Prognostic miR-146b MicroRNAs Has a Minimal and Negative Effect on the Malignant Phenotype of A549 Lung Cancer Cells