BACKGROUND AND AIM: Survivin has functions in both the regulation of cell division and the inhibition of apoptosis, and it is expressed in most human tumors but not in normal adult tissues. It is a potential target for cancer gene therapy. In the present study, we designed a DNAzyme targeting human survivin mRNA and tried to determine its effect on human pancreatic carcinoma cell line PANC-1. METHODS: We designed and synthesized a '10-23' antisurvivin mRNA DNAzyme, testified its cleavage activity by cell-free test, then delivered it into the PANC-1 cell line through liposome, and detected its effect on survivin mRNA expression by reverse transcription polymerase chain reaction, Western blot methods, evaluated its influence on the growth of PANC-1 cells by apoptosis detection, growth curve, and flow cytometry. RESULTS: Our results showed that the DNAzyme digested mRNA substrates of survivin efficiently in a dosage- and time-dependent manner, markedly increased apoptosis and inhibited the growth of human pancreatic carcinoma cell line PANC-1, compared with the disabled DNAzyme and untreated controls. CONCLUSIONS: Our results showed that the designed DNAzyme against survivin mRNA is a good candidate for cancer gene therapy of human pancreatic carcinoma.
BACKGROUND AND AIM: Survivin has functions in both the regulation of cell division and the inhibition of apoptosis, and it is expressed in most humantumors but not in normal adult tissues. It is a potential target for cancer gene therapy. In the present study, we designed a DNAzyme targeting human survivin mRNA and tried to determine its effect on humanpancreatic carcinoma cell line PANC-1. METHODS: We designed and synthesized a '10-23' antisurvivin mRNA DNAzyme, testified its cleavage activity by cell-free test, then delivered it into the PANC-1 cell line through liposome, and detected its effect on survivin mRNA expression by reverse transcription polymerase chain reaction, Western blot methods, evaluated its influence on the growth of PANC-1 cells by apoptosis detection, growth curve, and flow cytometry. RESULTS: Our results showed that the DNAzyme digested mRNA substrates of survivin efficiently in a dosage- and time-dependent manner, markedly increased apoptosis and inhibited the growth of humanpancreatic carcinoma cell line PANC-1, compared with the disabled DNAzyme and untreated controls. CONCLUSIONS: Our results showed that the designed DNAzyme against survivin mRNA is a good candidate for cancer gene therapy of humanpancreatic carcinoma.
Authors: Ravinay Bhindi; Roger G Fahmy; Harry C Lowe; Colin N Chesterman; Crispin R Dass; Murray J Cairns; Edward G Saravolac; Lun-Quan Sun; Levon M Khachigian Journal: Am J Pathol Date: 2007-08-23 Impact factor: 4.307