[1]唐艳,熊正爱,李攀,等.超声联合微泡造影剂携p53基因转染宫颈癌HeLa细胞的实验研究[J].第三军医大学学报,2009,31(23):2323-2326.
 TANG Yan,XIONG Zheng-ai,LI Pan,et al.Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier[J].J Third Mil Med Univ,2009,31(23):2323-2326.
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超声联合微泡造影剂携p53基因转染宫颈癌HeLa细胞的实验研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第23期
页码:
2323-2326
栏目:
论著
出版日期:
2009-12-15

文章信息/Info

Title:
Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier
作者:
唐艳熊正爱李攀王志刚董培婷
重庆医科大学附属第二医院:妇产科,超声影像研究所
Author(s):
TANG YanXIONG Zheng-aiLI Pan WANG Zhi-gangDONG Pei-ting
Department of Obstetrics and Gynecology, Institute of Ultrasonography, Second Affiliated Hospital,Chongqing Medical University,Chongqing 400010,China
关键词:
超声微泡造影剂宫颈癌基因治疗
Keywords:
ultrasound microbubble contrast agent cervical cancer gene therapy
分类号:
R454.3;R73-36;R737.33
文献标志码:
A
摘要:
目的    探讨超声联合微泡造影剂介导野生型p53(wtp53)基因转染宫颈癌HeLa细胞的可行性、效率性及作用效果。    方法    以前期实验所筛选的超声辐照参数(300 kHz ,0.5 W/cm2 , 30 s)为实验条件,将HeLa细胞分为质粒组(A组)、质粒+超声组(B组)、质粒+超声+微泡组(C组)、质粒+脂质体组(D组)、空白对照组(E组),分别进行处理。转染24~48 h后,收集各组细胞,荧光显微镜观察细胞基因转染效率、RT-PCR检测p53 mRNA表达,流式细胞仪检测细胞周期,MTT检测细胞增殖抑制情况。    结果    荧光显微镜下见C、D组均有较多绿色荧光蛋白表达的细胞(转染率分别为14.15%和10.86%),B组仅有极少的荧光细胞(0.81%);A组和E组无荧光表达;RT-PCR结果显示,C组和D组均可见特异性p53电泳条带,C组的电泳条带的光密度比值高于D组(P<0.05);流式细胞仪测细胞周期显示,转染野生型p53基因能使细胞周期出现明显的G1期阻滞,C、D组与E组相比差异有显著性(P<0.05);MTT测定结果显示C组细胞生长明显受抑,且随时间的延长,抑制程度逐渐增强。    结论    适当浓度的微泡和优化的超声辐照条件可增强基因转染效率,野生型p53基因能使HeLa细胞产生G1期阻滞,抑制宫颈癌细胞生长。
Abstract:
Objective    To explore the feasibility, efficiency and effect of wide-type p53 gene transfected into cervical cancer HeLa cells by ultrasound microbubble intensifier.     Methods    Based on the  optimum parameter of ultrasound wave irradiation (0.5 W/cm2 , 30 s) selected in preexperiment, HeLa cells were divided into 5 groups: (A)simple plasmid group, (B)plasmid-ultrasound group, (C)plasmid-ultrasound-microbubble group, (D)plasmid-liposome group, (E)blank control group. At 24-48h after transfection, the transfection efficiency was detected by fluorescence microscope, the mRNA expression of p53 was detected by RT-PCR, the cell cycle was detected by flow cytometry, and the inhibition rate of cells growth was detected by MTT assay.     Results    Fluorescence microscopy showed that the expression of green fluorescent protein in group C was higher than that in group Band group D (P<0.05), and there were no green fluorescent cells in group A and group E. RT-PCR confirmed that the specific p53 electrophoresis strips were seen in group C and group D. The optical density of the strip from group C was higher than that from group D (P<0.05). The cell cycles of transfected p53 cells arrested from G1 to S phase and the growth of cells transfected with wtp53 gene were dramatically inhibited. The characteristic of cells in group C and D was quite different from that of group E (P<0.05).     Conclusion    The transfection efficiency of foreign p53 gene into HeLa cells can be enhanced by optimum ultrasound intensity and microbubble dosage. Wtp53 gene can get the cell cycle arrested in G1 phase and inhibit the growth of  HeLa cells.

参考文献/References:

唐艳,熊正爱,李攀,等.超声联合微泡造影剂携p53基因转染宫颈癌HeLa细胞的实验研究[J]. 第三军医大学学报,2009,31(23):2323-2326.

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更新日期/Last Update: 2009-11-23