In vitro and in vivo double-enhanced suicide gene therapy mediated by generation 5 polyamidoamine dendrimers for PC-3 cell line.
BACKGROUND
One of the most frequently used and efficient suicide gene therapies for prostate cancer is HSV-TK/GCV system, but its application has been limited due to lack of favorable gene vector and the reduction of "bystander effect". We investigated the effect of a novel combination of HSV-TK/GCV fused with Cx43 and gemcitabine using non-viral vector generation 5 polyamidoamine dendrimers (G5-PAMAM-D) on PC-3 cells.


METHODS
RT-PCR and Western blot were used to detect TK and Cx43 expression. Cell viability and proliferation were measured by using MTT assay. Cell apoptosis was detected with double-staining of Annexin V-FITC and propidium iodide (PI) by flow cytometry. Nude mice models were established to evaluate the therapeutic effect in vivo.


RESULTS
G5-PAMAM-D efficiently delivered recombinant plasmids into PC-3 cells and HSV-TK and Cx43 could be expressed successfully. With gemcitabine, G5-PAMAM-D mediated HSV-TK and Cx43 expression effectively inhibited prostate cancer PC-3 cell proliferation, leading to more cellular apoptosis and inhibiting PC-3 tumor growth in nude mice models.


CONCLUSIONS
This study illustrates that this new suicide gene system mediated by G5-PAMAM-D is effective in decreasing PC-3 cell proliferation and inducing cell apoptosis, and inhibiting tumor growth in vivo. In a word, our study could provide a potential approach for gene therapy of prostate cancer.
Department of Urology, Second Hospital of TianJin Medical University, TianJin Institute of Urology, Tianjin, China.
Yue
Y
Chen
Gang
G
Wang
Deling
D
Kong
Zhihong
Z
Zhang
Kuo
K
Yang
Ranlu
R
Liu
Weiming
W
Zhao
Yong
Y
Xu
D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use; D017209:Apoptosis; D024201:Bystander Effect; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018031:Connexin 43 / Q000235:genetics* / Q000378:metabolism; D050091:Dendrimers / Q000494:pharmacology*; D015774:Ganciclovir / Q000627:therapeutic use; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010957:Plasmids / Q000235:genetics; D011073:Polyamines / Q000737:chemistry*; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
D000998:Antiviral Agents; D018031:Connexin 43; D050091:Dendrimers; C531249:Poly(amidoamine); D011073:Polyamines; D013937:Thymidine Kinase; D015774:Ganciclovir