Efficient gene delivery to myocardium with ultrasound targeted microbubble destruction and polyethylenimine

Chen, Zhiyi; Xie, Mingxing; Wang, Xinfang; Lv, Qing; Ding, Shan

doi:10.1007/s11596-008-0528-4

Cited by 16 publications

(17 citation statements)

References 21 publications

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“…Plasmid DNA (100 µl) or SonoVue/DNA complexes were added into 6 wells of 96-well culture plates, as described previously (15), and exposed to ultrasound irradiation (0.4, 1.0, 1.6 and 2.2 W/cm 2 ; DC, 20%; exposure time, 3 min) with or without the addition of SonoVue microbubbles. Gel electrophoresis analysis was performed immediately after ultrasound irradiation.…”

Section: Stability Of Plasmid Dna Following Ultrasound Exposurementioning

confidence: 99%

“…DNA was extracted from detached and adherent cells, and was subjected to electrophoresis using agarose gel as previously described (15).…”

Section: Reverse Transcription Pcr (Rt-pcr)mentioning

confidence: 99%

“…The use of a viral vector has been developed as a highly efficient method for gene delivery to a variety of tissues, although it evokes specific immune responses that may limit clinical application. Among non-viral techniques, ultrasound-targeted microbubble destruction (UTMD) has evolved as a new, promising tool for site-specific drug and gene delivery in vitro and in vivo, targeting delivery via a process called sonoporation, allowing for direct transfer into the cells (14)(15)(16). Significant efforts have been made to demonstrate the application of siRNA mediated by UTMD to block gene expression in vitro and in vivo (17)(18)(19)(20).…”

Section: Introductionmentioning

confidence: 99%

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Optimization and apoptosis induction by RNAi with UTMD technology in vitro

et al. 2012

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Abstract. Apoptosis induction by short hairpin RNA (shRNA) expression vectors may be an efficient and promising strategy for cancer gene therapy. Ultrasound-targeted microbubble destruction (UTMD) is an appealing technique; however, there few data are available to demonstrate the feasibility and to optimize the methodology for this technology. The aim of this study was to optimize this technique and to elucidate the effects on gene inhibition and apoptosis induction in vitro. Human cervical cancer cell lines were obtained and cultured. shRNA vectors were constructed, and the UTMD technique was examined to determine whether or not it was suitable for shRNA transfection into cells. Cells were then examined using flow cytometry. The results revealed that the optimal irradiation parameters obtained higher transfection efficiency and did not affect the integrity of plasmid DNA. We concluded that survivin downregulation with shRNA expression vectors, mediated by the optimal UTMD parameters, markedly induced cell apoptosis and caused cell cycle arrest, laying a foundation for further investigation of this cancer therapy.

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Section: Stability Of Plasmid Dna Following Ultrasound Exposurementioning

confidence: 99%

“…DNA was extracted from detached and adherent cells, and was subjected to electrophoresis using agarose gel as previously described (15).…”

Section: Reverse Transcription Pcr (Rt-pcr)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Optimization and apoptosis induction by RNAi with UTMD technology in vitro

et al. 2012

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“…Numerous studies have demonstrated delivery of reporters to myocardium. In general, most studies have confirmed that the ultrasound energy must be applied to the microbubble for payload delivery [76-78]; that is, delivery outside the ultrasound beam or mediated by ultrasound alone (without microbubbles) occurs at a very low rate. However, early work by [30] and [79] found transgene expression in distant (non-sonoporated organs), although this may be due to the use of a viral payload in the case of the latter.…”

Section: In Vivo Nucleic Acid Delivery With Sonoporation: Tissues mentioning

confidence: 99%

Nucleic acid delivery with microbubbles and ultrasound

Rychak

Klibanov

2014

Advanced Drug Delivery Reviews

107

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Nucleic acid-based therapy is a growing field of drug delivery research. Although ultrasound has been suggested to enhance transfection decades ago, it took a combination of ultrasound with nucleic acid carrier systems (microbubbles, liposomes, polyplexes, viral carriers) to achieve reasonable nucleic acid delivery efficacy. Microbubbles serve as foci for local deposition of ultrasound energy near the target cell, and greatly enhance sonoporation. Major advantage of this approach is in the minimal transfection in the non-insonated non-target tissues. Microbubbles can be simply co-administered with the nucleic acid carrier or can be modified to carry nucleic acid themselves. Liposomes with embedded gas or gas precursor particles can also be used to carry nucleic acid, release and deliver it by the ultrasound trigger. Successful testing in a wide variety of animal models (myocardium, solid tumors, skeletal muscle, pancreas) proves the potential usefulness of this technique for nucleic acid drug delivery.

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“…Several previous reports (16)(17)(18) have described the combined use of sonoporation and microbubbles to achieve delivery of naked plasmid DNA into cancer cells in tissue culture-based systems. Sonoporation is an emerging and promising physical method for cancer gene therapy that typically operates at a frequency of 1-35 MHz, and has several advantages over other nonphysical methods of nucleic acid delivery; for example, sonoporation also has the ability to deliver drugs or small molecules (19).…”

Section: Introductionmentioning

confidence: 99%

Liposome-mediated transfection of wild-type P53 DNA into human prostate cancer cells is improved by low-frequency ultrasound combined with microbubbles

Bai

Zhang

et al. 2016

Oncology Letters

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Abstract. Prostate cancer is a common type of cancer in elderly men. The aim of the present study was to evaluate the effects of ultrasound exposure in combination with SonoVue microbubbles on liposome-mediated transfection of wild-type P53 genes into human prostate cancer cells. PC-3 human prostate cancer cells were exposed to ultrasound; duty cycle was controlled at 20% (2 sec on, 8 sec off) for 5 min with and without SonoVue microbubble echo-contrast agent using a digital sonifier (frequency, 21 kHz; intensity, 46 mW/cm 2 ). The cells were divided into eight groups, as follows: Group A (SonoVue + wild-type P53), group B (ultrasound + wild-type P53), group C (SonoVue + ultrasound + wild-type P53), group D (liposome + wild-type P53), group E (liposome + SonoVue + wild-type P53), group F (liposome + wild-type P53 + ultrasound), group G (liposome + wild-type P53 + ultrasound + SonoVue) and the control group (wild-type P53). Following treatment, a hemocytometer was used to measure cell lysis, reverse transcription-quantitative polymerase chain reaction and western blotting were performed to detect P53 gene transfection efficiency, Cell Counting Kit-8 was employed to reveal cell proliferation and Annexin V/propidium iodide staining was used to determine cell apoptosis. Cell lysis was minimal in each group. Wild-type P53 gene and protein expression were significantly increased in the PC-3 cells in group G compared with the control and all other groups (P<0.01). Cell proliferation was significantly suppressed in group G compared with the control group and all other groups (P<0.01). Cell apoptosis levels in group G were significantly improved compared with the control group and all other groups (P<0.01). Thus, the results of the present study indicate that the use of low-frequency and low-energy ultrasound in combination with SonoVue microbubbles may be a potent physical method for increasing liposome gene delivery efficiency.

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Efficient gene delivery to myocardium with ultrasound targeted microbubble destruction and polyethylenimine

Cited by 16 publications

References 21 publications

Optimization and apoptosis induction by RNAi with UTMD technology in vitro

Optimization and apoptosis induction by RNAi with UTMD technology in vitro

Nucleic acid delivery with microbubbles and ultrasound

Liposome-mediated transfection of wild-type P53 DNA into human prostate cancer cells is improved by low-frequency ultrasound combined with microbubbles

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