Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier

唐艳; 熊正爱; 李攀; 王志刚; 董培婷

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Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier

更新时间：2026-01-09

- Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier
- Journal of Army Medical University Vol. 31, Issue 23, Pages: 2323-2326(2009)
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  CLC： R454.3;R73-36;R737.33
- Published：2009
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唐艳,熊正爱,李攀,王志刚,董培婷.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].陆军军医大学学报(原第三军医大学学报),2009,31(23):2323-2326.

　TANG Yan,XIONG Zheng-ai,LI Pan,WANG Zhi-gang,DONG Pei-ting.Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(23):2323-2326.
唐艳,熊正爱,李攀,王志刚,董培婷.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].陆军军医大学学报(原第三军医大学学报),2009,31(23):2323-2326. DOI：

　TANG Yan,XIONG Zheng-ai,LI Pan,WANG Zhi-gang,DONG Pei-ting.Experimental study of p53 plasmid transfected into HeLa cells by ultrasound microbubble intensifier[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(23):2323-2326. DOI：

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/cm

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（P0.05）；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G

期阻滞，C、D组与E组相比差异有显著性（P0.05）；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G

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/cm

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 gr

oup

(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 (P0.05)

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 (P0.05). The cell cycles of transfected p53 cells arrested from G

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 (P0.05). 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 G

phase and inhibit the growth of HeLa cells.

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