Feasibility and effect of ultrasound microbubble-mediated wild-type p53 gene transfection of HeLa cells

Chen,Wen-Juan; Xiong,Zheng-Ai; Tang,Yan; Dong,Pei-Ting; Li,Pan; Wang,Zhi-Gang

doi:10.3892/etm.2012.528

Feasibility and effect of ultrasound microbubble-mediated wild-type p53 gene transfection of HeLa cells

Authors:
- Wen-Juan Chen
- Zheng-Ai Xiong
- Yan Tang
- Pei-Ting Dong
- Pan Li
- Zhi-Gang Wang
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Affiliations: Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China, Institute of Ultrasound Imaging, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China
Published online on: March 27, 2012 https://doi.org/10.3892/etm.2012.528
Pages: 999-1004

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Abstract

Gene therapy holds great promise for the treatment of diseases. The key problem of gene therapy is the choice of an effective vector. Ultrasound-mediated microbubble technique (UMMT) has already shown promising applications in numerous types of tumors apart from cervical carcinoma. In the present study, according to the results of an MTT assay, we initially chose an ultrasound intensity of 0.5 W/cm2, an ultrasound exposure time of 30 sec and a microbubble concentration of 10% as the optimum experimental condition for wtp53 plasmid transfection into HeLa cells. To further investigate the transfection efficiency of ultrasound combined with microbubbles, RT-PCR analysis was used to examine the mRNA level of p53. The transfection efficiency in the plasmid plus microbubbles and ultrasound group was significantly higher than that of the other groups. Following transfection of the wtp53 gene, flow cytometric analysis showed that the cell cycle of HeLa cells was arrested in the G1 phase. The results of the present study suggest that UMMT, a new gene delivery system, increases the transfection efficiency of the wtp53 gene. Moreover, the growth of HeLa cells was arrested by introducing wtp53. This study may afford a new trend for the gene therapy of cervical carcinoma.

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1.	Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar
2.	Green J, Kirwan J, Tierney J, Vale C, Symonds P, Fresco L, Williams C and Collingwood M: Concomitant chemotherapy and radiation therapy for cancer of the uterine cervix. Cochrane Database Syst Rev. CD0022252005.
3.	Nishizaki M, Meyn RE, Levy LB, Atkinson EN, White RA, Roth JA and Ji L: Synergistic inhibition of human lung cancer cell growth by adenovirus-mediated wild-type p53 gene transfer in combination with docetaxel and radiation therapeutics in vitro and in vivo. Clin Cancer Res. 7:2887–2897. 2001.PubMed/NCBI
4.	Oki E, Tokunaga E, Nakamura T, et al: Genetic mutual relationship between PTEN and p53 in gastric cancer. Cancer Lett. 227:33–38. 2005. View Article : Google Scholar : PubMed/NCBI
5.	Lu QL, Liang HD, Partridge T and Blomley MJ: Microbubble ultrasound improves the efficiency of gene transduction in skeletal muscle in vivo with reduced tissue damage. Gene Ther. 10:396–405. 2003. View Article : Google Scholar : PubMed/NCBI
6.	Chen S, Ding JH, Bekeredjian R, et al: Efficient gene delivery to pancreatic islets with ultrasonic microbubble destruction technology. Proc Natl Acad Sci USA. 103:8469–8474. 2006. View Article : Google Scholar : PubMed/NCBI
7.	Oberle V, de Jong G, Drayer JI and Hoekstra D: Efficient transfer of chromosome-based DNA constructs into mammalian cells. Biochim Biophys Acta. 1676:223–230. 2004. View Article : Google Scholar : PubMed/NCBI
8.	Shohet RV, Chen S, Zhou YT, Wang Z, Meidell RS, Unger RH and Grayburn PA: Echocardiographic destruction of albumin microbubbles directs gene delivery to the myocardium. Circulation. 101:2554–2556. 2000. View Article : Google Scholar : PubMed/NCBI
9.	Xing W, Gang WZ, Yong Z, Yi ZY, Shan XC and Tao RH: Treatment of xenografted ovarian carcinoma using paclitaxel-loaded ultrasound microbubbles. Acad Radiol. 15:1574–1579. 2008. View Article : Google Scholar : PubMed/NCBI
10.	Chen Z, Xie M, Wang X, Lv Q and Ding S: Efficient gene delivery to myocardium with ultrasound targeted microbubble destruction and polyethylenimine. J Huazhong Univ Sci Technolog Med Sci. 28:613–617. 2008. View Article : Google Scholar : PubMed/NCBI
11.	Bekeredjian R, Kroll RD, Fein E, et al: Ultrasound targeted microbubble destruction increases capillary permeability in hepatomas. Ultrasound Med Biol. 33:1592–1598. 2007. View Article : Google Scholar : PubMed/NCBI
12.	Anwer K, Kao G, Proctor B, et al: Ultrasound enhancement of cationic lipid mediated gene transfer to primary tumors following systemic administration. Gene Ther. 7:1833–1839. 2000. View Article : Google Scholar : PubMed/NCBI
13.	Nie F, Xu HX, Tang Q and Lu MD: Microbubble-enhanced ultra-sound exposure improves gene transfer in vascular endothelial cells. World J Gastroenterol. 12:7508–7513. 2006.PubMed/NCBI
14.	Tomkova K, Tomka M and Zajac V: Contribution of p53, p63, and p73 to the developmental diseases and cancer. Neoplasma. 55:177–181. 2008.PubMed/NCBI
15.	Nork TM, Poulsen GL, Millecchia LL, Jantz RG and Nickells RW: p53 regulates apoptosis in human retinoblastoma. Arch Ophthalmol. 115:213–219. 1997. View Article : Google Scholar : PubMed/NCBI
16.	Ghule P, Kadam PA, Jambhekar N, et al: p53 gene gets altered by various mechanisms: studies in childhood sarcomas and retinoblastoma. Med Sci Monit. 12:BR385–BR396. 2006.
17.	Xu L, Pirollo KF, Tang WH, Rait A and Chang EH: Transferrin liposome mediated systemic p53 gene therapy in combination with radiation results in regression of human head and neck cancer xenografts. Hum Gene Ther. 10:2941–2952. 1999. View Article : Google Scholar : PubMed/NCBI
18.	Haupt S and Haupt Y: Importance of p53 for cancer onset and therapy. Anticancer Drugs. 17:725–732. 2006. View Article : Google Scholar : PubMed/NCBI
19.	Tiukinhoy SD, Mahowald ME, Shively VP, et al: Development of echogenic, plasmid-incorporated, tissue-targeted cationic liposomes that can be used for directed gene delivery. Invest Radiol. 35:732–738. 2000. View Article : Google Scholar
20.	Li T, Tachibana K and Kuroki M and Kuroki M: Gene transfer with echo-enhanced contrast agents: comparison between Albunex, Optison, and Levovist in mice-initial results. Radiology. 229:423–428. 2003. View Article : Google Scholar : PubMed/NCBI
21.	Shimamura M, Sato N, Taniyama Y, et al: Development of efficient plasmid DNA transfer into adult rat central nervous system using microbubble-enhanced ultrasound. Gene Ther. 11:1532–1539. 2004. View Article : Google Scholar : PubMed/NCBI
22.	Zhigang W, Zhiyu L, Haitao R, et al: Ultrasound-mediated microbubble destruction enhances VEGF gene delivery to the infarcted myocardium in rats. Clin Imaging. 28:395–398. 2004. View Article : Google Scholar : PubMed/NCBI
23.	Zhang Q, Wang Z, Ran H, et al: Enhanced gene delivery into skeletal muscles with ultrasound and microbubble techniques. Acad Radiol. 13:363–367. 2006. View Article : Google Scholar : PubMed/NCBI
24.	Mitragotri S, Blankschtein D and Langer R: Transdermal drug delivery using low-frequency sonophoresis. Pharm Res. 13:411–420. 1996. View Article : Google Scholar : PubMed/NCBI
25.	Mitragotri S and Kost J: Low-frequency sonophoresis: a noninvasive method of drug delivery and diagnostics. Biotechnol Prog. 16:488–492. 2000. View Article : Google Scholar : PubMed/NCBI
26.	Tachibana K, Uchida T, Tamura K, Eguchi H, Yamashita N and Ogawa K: Enhanced cytotoxic effect of Ara-C by low intensity ultrasound to HL-60 cells. Cancer Lett. 149:189–194. 2000. View Article : Google Scholar : PubMed/NCBI
27.	Munshi N, Rapoport N and Pitt WG: Ultrasonic activated drug delivery from Pluronic P-105 micelles. Cancer Lett. 118:13–19. 1997. View Article : Google Scholar : PubMed/NCBI
28.	Yu T, Hu K, Bai J and Wang ZB: Reversal of adriamycin resistance in ovarian carcinoma cell line by combination of verapamil and low-level ultrasound. Ultrason Sonochem. 10:37–40. 2003. View Article : Google Scholar : PubMed/NCBI
29.	Dalecki D, Raemen CH, Child SZ, Cox C, Francis CW, Meltzer RS and Carstensen EL: Hemolysis in vivo from exposure to pulsed ultrasound. Ultrasound Med Biol. 23:307–313. 1997. View Article : Google Scholar : PubMed/NCBI