The culture induced senescence of mouse embryonic fibroblasts (MEF) correlates with reduction of cell proliferation. In this work we found that the accumulation of cells with 4C DNA content and the transcriptional change of several miRNAs are relevant events in culture senescence. By comparing the miRNA expression profiles of physiologically senescent MEF and that of senescent MEF induced by the down-regulation of Leukemia Related factor (LRF), miR-290 was identified as a common up-regulated miRNA. When miR-290 was transfected in pre-senescent MEF, SA--gal⁺ cells and p16, two markers of culture senescence, increased in comparison to control indicating that miR-290 is causally involved in senescence. Interestingly Nocodazole (NCZ), which induces G2/M block, increased the percentage of senescent cells as well as the expression of miR-290 and of the tumor suppressor p16, thus mimicking culture senescence. As miR-290 was over-expressed in NCZ treated cells and it was able to induce senescence in proliferating MEF, we investigated whether miR-290 and NCZ could share common mechanisms of culture senescence. Whereas the induction of SA--gal⁺ by miR-290 was not strengthened by coupling its transfection with NCZ treatment, the transfection of the antagomir d-290 plus NCZ treatment, while blocking cells at G2/M, suppressed SA--gal⁺ and p16 induction. On the basis of these findings we conclude that miR-290 might act as a physiological effector of NCZ induced as well as culture senescence via p16 regulation expanding the role of this miRNA from embryonic stem to differentiated cells.