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纸质出版:2012
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谢鑫,姜庆,梁培禾,王志刚,郑元义,谭小军,乔娟.携HSV1-TK自杀基因超声微泡造影剂联合前药更昔洛韦对前列腺癌细胞的抑制[J].陆军军医大学学报(原第三军医大学学报),2012,34(13):1285-1288.
Xie Xin,Jiang Qing,Liang Peihe,Wang Zhigang,Zheng Yuanyi,Tan Xiaojun,Qiao Juan.Inhibitory effect of ultrasound microbubbles carrying HSV1-TK gene combined with ganciclovir on prostate cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(13):1285-1288.
谢鑫,姜庆,梁培禾,王志刚,郑元义,谭小军,乔娟.携HSV1-TK自杀基因超声微泡造影剂联合前药更昔洛韦对前列腺癌细胞的抑制[J].陆军军医大学学报(原第三军医大学学报),2012,34(13):1285-1288. DOI:
Xie Xin,Jiang Qing,Liang Peihe,Wang Zhigang,Zheng Yuanyi,Tan Xiaojun,Qiao Juan.Inhibitory effect of ultrasound microbubbles carrying HSV1-TK gene combined with ganciclovir on prostate cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(13):1285-1288. DOI:
目的 观察超声微泡造影剂携单纯疱疹病毒Ⅰ型胸昔激酶(HSV
1
-TK)基因体外转染及联合前药更昔洛韦(GCV)对前列腺癌细胞株PC-3的抑制效应。 方法 超声辐照PC-3细胞,MTT筛选出最适辐照时间;将含有HSV
1
-TK基因的质粒通过静电吸附在脂质微泡表面,采用最适超声辐照转染,并用荧光显微镜及Western blot观察TK基因的转入及表达;MTT法观察使用不同浓度的前药更昔洛韦(GCV)后细胞增殖抑制率;流式细胞仪检测最适浓度的前药GCV对转染后的前列腺癌细胞PC-3的周期的影响;MTT法观察HSV
1
-TK/GCV自杀基因系统对前列腺癌PC-3细胞杀伤效应。 结果 MTT检测显示超声强度为1.0 W/cm
2
频率为1 MHz,辐照时间30 s时超声辐照对细胞无明显的抑制作用;通过超声辐照后HSV
1
-TK基因可以顺利转入PC-3细胞中并有稳定的表达。与对照组相比,当前药GCV浓度在100 μg/ml时,超声+微泡+TK组细胞杀伤率明显高于其他各组,细胞存活率约32%,低于其他各组(P0.05);流式细胞术检测显示大多数细胞被阻断在S期,细胞生长受到明显抑制。 结论 以超声辐照微泡造影剂为载体转染自杀基因联合前药GCV,对人前列腺癌细胞有明显杀伤作用。
Objective To determine the effect of transfection of ultrasonic microbubble with herpes simplex virus thymidine kinase (HSV
1
-TK) gene on prostate cancer cell line PC-3 combined with ganciclovir (GCV). Methods MTT assay was used to test the optimal irradiation time of PC-3 cells by ultrasonic irradiation. Ultrasound microbubble surface was adsorbed with HSV
1
-TK gene
and then tran
sfected into PC-3 cells under optimal ultrasound irradiation. Fluorescence microscopy and Western blot analysis were used to detect the transfection and expression of TK. The cell proliferation was measured by MTT assay after the treatment of different concentrations of GCV. Flow cytometry (FCM) was used to measure the cell cycle of the transfected PC-3 cells after the treatment of the optimal concentration of GCV. The killing effect of HSV
1
-TK/GCV suicide gene system on prostate cancer cell PC-3 was observed by MTT assay. Results MTT assay showed that ultrasonic radiation had no significant inhibition on cell viability under the conditions of the magnitude of 1.0 W/cm
2
frequency of 1 MHz
and every 30 s ultrasound exposure time. HSV
1
-TK was transfected into PC-3 cells by ultrasound wave irradiation and expressed stably. Compared with the control
the killing activity of ultrasound radiation+ microbubble + TK treatment with the GCV concentration of 100 μg/ml was obviously higher
with cell survival rate of about 32%
significantly lower than the other group (P0.05). FCM showed that most cells were blocked in S phase
and cell growth was obviously inhibited. Conclusion Transfection of suicide gene by ultrasound microbubble and the application of GCV has an obvious effect on the apoptosis of human prostate cancer cells.
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