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Effect of targeted ovarian cancer therapy using amniotic fluid mesenchymal stem cells transfected with enhanced green fluorescent protein-human interleukin-2 in vivo.

You Q, Yao Y, Zhang Y, Fu S, Du M, Zhang G - Mol Med Rep (2015)

Bottom Line: The aim of the present study was to investigate the effect of using amniotic fluid mesenchymal stem cells (AF-MSCs) in targeted ovarian cancer therapy in vivo.AF-MSCs were isolated from human second trimester AF and a plasmid, enhanced green fluorescent protein‑human interleukin‑2 (pEGFP‑hIL‑2) was formed.It was found that AF‑MSCs exhibited high motility during migration in vivo, and the vector, pEGFP‑hIL‑2 can be stably transfected into AF‑MSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Gynecology and Obstetrics, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.

ABSTRACT
The aim of the present study was to investigate the effect of using amniotic fluid mesenchymal stem cells (AF-MSCs) in targeted ovarian cancer therapy in vivo. AF-MSCs were isolated from human second trimester AF and a plasmid, enhanced green fluorescent protein‑human interleukin‑2 (pEGFP‑hIL‑2) was formed. The plasmid was stably transfected into the AF‑MSCs and the cells were intravenously injected into ovarian cancer nude mice models. Following stable transfection of the vector, tumor formation, and the expression and activity of hIL‑2 were investigated, and microscopic pathological examinations of the tumor were performed. It was found that AF‑MSCs exhibited high motility during migration in vivo, and the vector, pEGFP‑hIL‑2 can be stably transfected into AF‑MSCs. Following stable transfection, this type of stem cell is able to successfully transport the therapeutic gene, IL-2, migrate to the ovarian cancer tumor site to secrete the functional IL-2 and treat the tumor. Thus, AF-MSCs may serve as transporters for therapeutic genes targeting ovarian tumor sites and, therefore, be involved in the treatment of tumors.

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Related in: MedlinePlus

Scheme depicting the plasmid pMD18-T vector which is linked with the interleukin-2 gene in the present study.
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f2-mmr-12-04-4859: Scheme depicting the plasmid pMD18-T vector which is linked with the interleukin-2 gene in the present study.

Mentions: The purified product was combined with a pMD18-T Simple Vector (Fig. 2) and 10 µl of the above-mentioned DNA product was mixed with 100 µl JM109 competent cells (provided by Dr Yin Zhe, Department of Microbiology, China Agricultural University, Harbin, China) in an ice bath. The mixture was placed in ice for 30 min, heated at 42°C for 90 sec and placed in an ice bath for a further 1–2 min. Following this, 800 µl medium without antibiotics was added into the above-mentioned sample and centrifuged at 22 × g for 40 min at 37°C. After centrifugation, ~100 µl of the supernatant was kept and spread on Luria-Bertani (LB) agar plates with Penbritin, X-gal and osopropyl β-D-1-thiogalactopyranoside (100 µg/ml of each). Twenty-four hours later, the LB mixture inoculated with Penbritin was agitated at 37°C for 8 h. The plasmid was extracted according to the manufacturer's instructions. Following PCR amplification, the sample was digested with EcoRI and SalI, and agarose gel electrophoresis was performed. The length of the DNA fragment obtained was 462 bp, which was verified by Invitrogen Life Technologies, and the recombinant plasmid was designated pMD-hIL-2.


Effect of targeted ovarian cancer therapy using amniotic fluid mesenchymal stem cells transfected with enhanced green fluorescent protein-human interleukin-2 in vivo.

You Q, Yao Y, Zhang Y, Fu S, Du M, Zhang G - Mol Med Rep (2015)

Scheme depicting the plasmid pMD18-T vector which is linked with the interleukin-2 gene in the present study.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581758&req=5

f2-mmr-12-04-4859: Scheme depicting the plasmid pMD18-T vector which is linked with the interleukin-2 gene in the present study.
Mentions: The purified product was combined with a pMD18-T Simple Vector (Fig. 2) and 10 µl of the above-mentioned DNA product was mixed with 100 µl JM109 competent cells (provided by Dr Yin Zhe, Department of Microbiology, China Agricultural University, Harbin, China) in an ice bath. The mixture was placed in ice for 30 min, heated at 42°C for 90 sec and placed in an ice bath for a further 1–2 min. Following this, 800 µl medium without antibiotics was added into the above-mentioned sample and centrifuged at 22 × g for 40 min at 37°C. After centrifugation, ~100 µl of the supernatant was kept and spread on Luria-Bertani (LB) agar plates with Penbritin, X-gal and osopropyl β-D-1-thiogalactopyranoside (100 µg/ml of each). Twenty-four hours later, the LB mixture inoculated with Penbritin was agitated at 37°C for 8 h. The plasmid was extracted according to the manufacturer's instructions. Following PCR amplification, the sample was digested with EcoRI and SalI, and agarose gel electrophoresis was performed. The length of the DNA fragment obtained was 462 bp, which was verified by Invitrogen Life Technologies, and the recombinant plasmid was designated pMD-hIL-2.

Bottom Line: The aim of the present study was to investigate the effect of using amniotic fluid mesenchymal stem cells (AF-MSCs) in targeted ovarian cancer therapy in vivo.AF-MSCs were isolated from human second trimester AF and a plasmid, enhanced green fluorescent protein‑human interleukin‑2 (pEGFP‑hIL‑2) was formed.It was found that AF‑MSCs exhibited high motility during migration in vivo, and the vector, pEGFP‑hIL‑2 can be stably transfected into AF‑MSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Gynecology and Obstetrics, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.

ABSTRACT
The aim of the present study was to investigate the effect of using amniotic fluid mesenchymal stem cells (AF-MSCs) in targeted ovarian cancer therapy in vivo. AF-MSCs were isolated from human second trimester AF and a plasmid, enhanced green fluorescent protein‑human interleukin‑2 (pEGFP‑hIL‑2) was formed. The plasmid was stably transfected into the AF‑MSCs and the cells were intravenously injected into ovarian cancer nude mice models. Following stable transfection of the vector, tumor formation, and the expression and activity of hIL‑2 were investigated, and microscopic pathological examinations of the tumor were performed. It was found that AF‑MSCs exhibited high motility during migration in vivo, and the vector, pEGFP‑hIL‑2 can be stably transfected into AF‑MSCs. Following stable transfection, this type of stem cell is able to successfully transport the therapeutic gene, IL-2, migrate to the ovarian cancer tumor site to secrete the functional IL-2 and treat the tumor. Thus, AF-MSCs may serve as transporters for therapeutic genes targeting ovarian tumor sites and, therefore, be involved in the treatment of tumors.

Show MeSH
Related in: MedlinePlus