ISSN 2329-9835
International Journal of Diseases and Disorders ISSN 2329-9835 Vol. 3 (3), pp. 001-007, March, 2015. © International Scholars Journals
Full Length Research Paper
Effect of shRNA-mediated survivin gene silencing on the biological behavior of human bladder transitional cell carcinoma T24 cells
Yunfeng He1, Hu Du1, Xiaohou Wu1*, Wei Tang1, Chuan Liu2, Zhikang Yin1, Yao Zhang1, Chunli Luo3, and Jiabing Li4
1Department of Urology, The First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
2Department of Urology, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
3Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
4The Third hospital of mianyang, No. 190 jiannian Road, mianyang; 621000 China.
Accepted 13 September, 2014
Abstract
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 survivin-specific 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.
Key words: Survivin, short hairpin RNA, Neoplasm invasiveness, bladder neoplasms.