MicroRNAs (miRNAs) are endogenous 19–25 nucleotide noncoding single-stranded RNAs that regulate gene expression by blocking the translation or decreasing the stability of mRNAs. In this study, with the treatment of CsA (Cyclosporin A), we showed that miR144 expression levels were decreased while titin mRNA expression levels were increased in human trophoblast (HT) cells, and identified titin as a novel direct target of miR-144. Overexpression of miR-144 suppressed titin and its downstream signaling molecule such as p-ERK1/2 and MMP2/9 expression, and attenuated cell proliferation and invasion. Forced expression of titin can partly rescue the inhibitory effect of miR-144 in the cells. Taken together, these findings will shed light to the role and mechanism of miR-144 in regulating HT cells proliferation and invasion via miR-144/titin axis, and miR-144 may serve as a potential therapeutic target in HT in the future. J. Cell. Biochem. 115: 690–696, 2014. © 2013 Wiley Periodicals, Inc.