Abstract: Objective To construct a plasmid expressing a small hairpin RNA（shRNA）to knockdown expression of human telomerase reverse transcriptase （hTERT），and to observe its effects on hTERT expression in caski cervical cancer cells. Methods Designer   3.0 software （Genepharma）was used to design an shRNA targeting hTERT. The shRNA was cloned into the pGLV3/Hi/GFP+Puro   vector and confirmed by sequencing. Cells were divided into blank control group，negative control group and hTERT interference   group. The resulting plasmid was transfected into caski cervical cancer cells and expression of the GFP reporter was analyzed. In   addition，expression of chromosomal hTERT mRNA was quantified using RT-PCR. Cell proliferation in the presence and absence of shRNA was measured using the cck-8 assay.Results A plasmid expressing an shRNA targeting hTERT was constructed and used to generate recombinant lentivirus. Lentiviral infection of caski cells led to lower expression of hTERT mRNA and slower cell   proliferation than in controls. Conclusion A plasmid expressing shRNA targeting hTERT can effectively knockdown endogenous   hTERT expression. This reagent may prove useful for understanding the role of telomerase in the pathogenesis of cervical cancer.

Key words: Cervical neoplasm, Human telomerase reverse transcriptase gene （hTERT）, Lentivirus-based vectors, Small hairpin RNA, Caski cell