Objective: Telomerase reactivation has been linked to cancer development and its biology. Loss of heterozygosity at chromosome 3p has been shown to correlate with telomerase activity, however, any gene in this region was not found to regulate hTERT expression. In here, we found miR-135a which is a novel regulator hTERT. Materials and Methods: Expression of miR-135a was measured with Taqman probe in 82 non-small cell lung cancers (NSCLCs). The methylation of miR-135a promoter was investigated by bisulfite methylation assay in 173 NSCLCs and lung cancer cell lines. miR-135a precursor and miR-135a inhibitor were transfected to H1299 cell to identify that it can regulate telomerase expression. Colony formation assay was used for the function of miR-135a in lung cancer tumorigenesis. Result: miR-135a was significantly downregulated in squamous cell carcinomas (p＜0.001) but not in adenocarcinomas. miR-135 down-regulation was correlated with genetic and epigenetic alteration of miR1-135. miR-135a precursor down-regulate hTERT expression through direct binding to its 3`UTR. Transfection of miR-135a in lung cancer cell significantly reduce colony formation ability than scramble miRNA and miR-135a inhibitor. Conclusions: These results suggest that loss of miR-135a expression may contribute to the gain of hTERT protein expression in lung tumorigenesis. miR-135a might play an important role in squamous cell carcinogenesis.