کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1269115 972440 2013 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Regulation of gene expression in human prostate cancer cells with artificially constructed promoters that are activated in response to ultrasound stimulation
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی (عمومی)
پیش نمایش صفحه اول مقاله
Regulation of gene expression in human prostate cancer cells with artificially constructed promoters that are activated in response to ultrasound stimulation
چکیده انگلیسی

We chose promoters responsive to sonication in LNCap cells, a prostate cancer cell line, out of a library composed of DNA fragments constructed by linking the TATA box sequence to randomly combined cis-acting elements of transcription factors activated in response to radiation in prostate cancer cells. When a plasmid containing the luciferase gene under control of a promoter was transfected into LNCap cells and sonicated with 1 MHz ultrasound at 0.5 W/cm2, 10% DF for 60 s, 13 promoters showed more than 10-fold enhancement compared with their counterparts without sonication 12 h after sonication. As to their responsiveness to sonication, the best two promoters were then compared to clone 880-8, a derivative from clone 880 that was created by random introduction of point mutations and was shown to have an improved response to X-ray irradiation. We then took clone 880-8 for further analyses since it showed the highest enhancement to sonication, though not statistically significant from the others. Next, we employed a retrovirus vector and stably introduced the luciferase gene under control of clone 880-8 into LNCap cells to establish a cell line. When the cell line was sonicated with 1 MHz ultrasound at 0.5 W/cm2, 10% DF for 60 s, luciferase expression was enhanced up to 14.8-fold 12 h after sonication. We then established another cell line by replacing the luciferase gene with the fcy::fur gene, a suicide gene, and when the cell line was sonicated with 1 MHz ultrasound at 0.5 W/cm2, 10% DF for 60 s, expression of the gene was enhanced, showing the maximum expression 12–24 h after sonication. When the cells were incubated in medium containing 5-fluorocytosine, cell survival ratio decreased dose dependently with 5-fluorocytosine only after sonication treatment, suggesting this promoter could be utilized for gene expression control with ultrasound.


► A promoter responsive to sonication was obtained of an artificially constructed promoter library.
► Expressions of genes connected downstream of the promoter were regulated with ultrasound.
► When a suicide gene was connected, in vitro suicide gene therapy could be controlled by ultrasound.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Ultrasonics Sonochemistry - Volume 20, Issue 1, January 2013, Pages 460–467
نویسندگان
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