CAJ | 학술논문

目的研究低频低能量超声联合微泡增强脂质体介导的野生型p53质粒转染促进人前列腺癌DU145细胞的早期细胞凋亡,及与p53-MDM2形成的负反馈环关系.方法以频率20 kHz、声功率为80 mW的超声,采用连续波辐照,实验分4组:空白对照组(A组)、单纯超声联合微泡组(B组)、脂质体转染组(C组)和超声加微泡联合脂质体转染组(D组),其中B、D组每毫升细胞悬液加微泡200μl C、D组每毫升细胞悬液加质粒8μg、脂质体20 μg,处理后细胞继续培养24 h,流式细胞仪检测细胞早期凋亡情况,36 h后Western blot检测细胞中p53蛋白及MDM2蛋白的表达量.结果流式结果显示D、C组的早期细胞凋亡率高于A、B组,差异有统计学意义(P＜0.01),以D组与A、B组的差异最明显,D组的早期细胞凋亡率亦高于C组,差异有统计学意义(P ＜0.01);Western blot显示D、C组p53表达较A、B组表达增加,同时MDM2的表达较A、B组表达减少,差异有统计学意义(P＜0.01),以D组变化最明显,D组与C组比较差异也有统计学意义(P ＜0.01).结论超声联合微泡促进脂质体介导的野生型p53质粒转染人雄激素非依赖型前列腺癌DU145细胞,可明显提高癌细胞的早期细胞凋亡率,这种重新表达野生型p53蛋白的DU145细胞早期细胞凋亡率的提高可能与上调p53蛋白、下调MDM2蛋白的表达有关.
목적 연구저빈저능량초성연합미포증강지질체개도적야생형p53질립전염촉진인전렬선암DU145세포적조기세포조망,급여p53-MDM2형성적부반궤배관계.방법 이빈솔20 kHz、성공솔위80 mW적초성,채용련속파복조,실험분4조:공백대조조(A조)、단순초성연합미포조(B조)、지질체전염조(C조)화초성가미포연합지질체전염조(D조),기중B、D조매호승세포현액가미포200μl C、D조매호승세포현액가질립8μg、지질체20 μg,처리후세포계속배양24 h,류식세포의검측세포조기조망정황,36 h후Western blot검측세포중p53단백급MDM2단백적표체량.결과 류식결과현시D、C조적조기세포조망솔고우A、B조,차이유통계학의의(P＜0.01),이D조여A、B조적차이최명현,D조적조기세포조망솔역고우C조,차이유통계학의의(P ＜0.01);Western blot현시D、C조p53표체교A、B조표체증가,동시MDM2적표체교A、B조표체감소,차이유통계학의의(P＜0.01),이D조변화최명현,D조여C조비교차이야유통계학의의(P ＜0.01).결론 초성연합미포촉진지질체개도적야생형p53질립전염인웅격소비의뢰형전렬선암DU145세포,가명현제고암세포적조기세포조망솔,저충중신표체야생형p53단백적DU145세포조기세포조망솔적제고가능여상조p53단백、하조MDM2단백적표체유관.
Objective To investigate whether the transfection of wild-type p53 gene into DU145 cells through 20 kHz ultrasound combined with microbubbles and liposome can induce cell apoptosis,and to explore the probable mechanism.Methods Ultrasound with a frequency of 20 kHz and power of 80 mW in continuous wave mode was used to transfect wild-type p53 gene into DU145 cells,and microbubble and liposome were added.DU145 cells were divided into four groups:control group (A),ultrasound combined with microbubbles group(B),liposome group(C),and ultrasound combined with microbubbles and liposome group(D).Twenty-four hours after treatment,the early apoptosis was detected by flow cytometry,and western blotting was used to evaluate the expression of the apoptosis-related proteins,p53 and MDM2,after treatment for thirty-six hours.Results The early apoptosis rate of group D was much higher than that of group A,B and C (P ＜0.01).Western blotting showed that the treatment of group D increase the expression of p53 protein and decreased the expression of MDM2 protein,which has a significance difference when compared with group A,B and C(P ＜0.01).Conclusions The treatment of transfection wild-type p53 gene into DU145 cells through 20 kHz ultrasound combined with microbubbles and liposome induce cell apoptosis may through up-regulation of p53 and down-regulation MDM2.