title: A preliminary study on the construction of double suicide gene delivery vectors by mesenchymal stem cells and the in vitro inhibitory effects on SKOV3 cells.
abstract: The aim of the present study was to investigate the efficacy of using human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) as gene delivery vectors in the treatment of ovarian cancer. Lentivectors overexpressing cytosine deaminase (CD) and herpes simplex virus thymidine kinase (HSV-tk) (pGC-FU-CD-TK) were constructed, and confirmed by enzyme digestion, DNA sequence and western blotting. Quantitative PCR (PCR) was used to verify the overexpression of the fusion gene (CD and HSV-tk). SKOV3 cells were co-cultured with MSCs/tk+CD+ at a 1:1 ratio, and were then treated with the prodrugs (GCV) and/or 5-fluorocytosine (5-FC) at different concentrations, and the cytotoxic effects were evaluated using MTT assay and flow cytometry. DNA sequencing demonstrated that the sequence of HSV-tk and CD genes were consistent with the objective sequence and western blotting verified that the constructed lentivector could produce the HSV-tk/CD gene. The packed titer was 2.00e+8 TU/ml. The pGC-FU-CD-TK could be stably transferred to hUCB‑MSCs, and the infection efficiency was almost 80%. RT-PCR demonstrated that the expression levels of the HSV-tk/CD fusion gene in MSCs/tk+CD+ group was 75 times that in the negative control (P<0.05). Compared with GCV or 5-FC alone, the growth inhibition rate (GIR) was significantly higher in the combined treatment (F=85.35, P<0.05). The reconstructed MSCs/tk+CD+ vectors were capable of slowing down the growth of human SKOV3 cells in the presence of prodrugs in vitro. The use of combination chemotherapy exhibited a more significant inhibitory effect than using a single prodrug.
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Jingyan
authors: J
authors: Jiang
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Dee
authors: D
authors: Wei
authors: Department of Gynecologic Oncology, Shandong Cancer Hospital, Jinan, Shandong 250117, P.R. China.
authors: Li
authors: L
authors: Sun
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Yuxia
authors: Y
authors: Wang
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Xihai
authors: X
authors: Wu
authors: Department of Stomatology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Ying
authors: Y
authors: Li
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Zhenghui
authors: Z
authors: Fang
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Hui
authors: H
authors: Shang
authors: Department of Gynecology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong 250014, P.R. China.
authors: Zengtao
authors: Z
authors: Wei
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000963:Antimetabolites / Q000494:pharmacology; D001483:Base Sequence; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D043525:Cytosine Deaminase / Q000096:biosynthesis / Q000235:genetics* / Q000627:therapeutic use; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D005437:Flucytosine / Q000494:pharmacology; D018014:Gene Transfer Techniques; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000628:therapy; D011355:Prodrugs / Q000627:therapeutic use; D011993:Recombinant Fusion Proteins; D017422:Sequence Analysis, DNA; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics* / Q000627:therapeutic use
chemical_list: D000963:Antimetabolites; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; D005437:Flucytosine; D013937:Thymidine Kinase; D043525:Cytosine Deaminase