Abstract

Yunfeng He, Hu Du, Xiaohou Wu*, Wei Tang, Chuan Liu, Zhikang Yin, Yao Zhang, Chunli Luo, and Jiabing Li

To construct the expression vector of the survivin-specific short hairpin RNA (shRNA) and to investigate its effect on the biological behavior of human bladder transitional cell carcinoma T24 cells. Survivin-specific short oligonucleotides were designed, synthesized, and cloned into the PTZU6+1 vector, resulting in a survivinspecific shRNA expression construct, which was then transfected into T24 cells. The mRNA and protein levels of survivin were determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Tumor cell growth, movement and neoplasm invasiveness were detected by a cell growth curve, a random motion experiment, and a Matrigel transmembrane assay. Target DNA fragments containing psurvivin-shRNA vectors was verified by EcoR I / Hind III double digestion and sequencing and successfully transfected into T24 cells. Transfection of psurvivin-shRNA cells significantly inhibited mRNA levels by 61.73% and protein expression by 73.37%. The ability of cell invasion and movement was significantly decreased. The inhibition rates of tumor cell growth were 59.13% on day 3 and 83.86% on day 5. A survivin-specific shRNA expression vector was constructed based on the PTZU6 +1 vector and was transfected into T24 cells, which resulted in the inhibition of T24 cell proliferation and movement.