摘要
目的探讨超声微泡介导自杀基因对卵巢癌SKOV3细胞抗瘤效应的机制。方法采用最适超声转染参数,以超声介导微泡转染pORF-HSV1TK质粒,将SKOV3细胞分成6组:阴性对照组(C组)、超声辐照组(U组)、脂质体组(L组,阳性对照组)、脂质体+超声辐照组(L+U组)、脂质体+微泡+超声辐照组(L+M+U组)和微泡+超声辐照组(M+U组)。以RT-PCR检测HSV1TK的表达,以MTT测定各组细胞增殖抑制率。光镜下观察各组转染细胞数量、形态变化,以流式细胞术(FCM)检测各组细胞凋亡率及细胞周期。结果L+M+U组HSV1TK基因表达最强,细胞增殖抑制率明显高于其他组(P均<0.05),且光镜下细胞数量减少最多,形态明显异常;其细胞凋亡率为(49.13±0.82)%,且大多数细胞周期被阻断于G1期(P<0.05)。结论超声介导微泡能增强HSV1TK/GCV系统抑制SKOV3增殖和诱导SKOV3凋亡的作用,且将SKOV3细胞周期阻断在G1期,从而发挥抗瘤效应。
Objective To explore the anti-tumor effect of ultrasound microbubbles mediated suicide gene on ovarian carcinoma SKOV3 cells.Methods Under the conditions of optimal ultrasound transfection parameters,pORF-HSV1TK plasmid was transfected into SKOV3 cells by ultrasound mediated microbubbles.Then the cells were divided into negative control group（group C）,ultrasound wave irradiation group（group U）,liposome group（group L,positive control group）,liposome and ultrasound wave irradiation group（group L＋U）,liposome,microbubbles and ultrasound wave irradiation group（group L＋M＋U） as well as microbubbles and ultrasound wave irradiation group（group M＋U）.The expression of HSV1TK was detected by using RT-PCR,the inhibition rates of proliferation of SKOV3 cells were detected by using MTT,the changes of quantity and morphology of SKOV3 cells was observed by using light microscope,and the rate of early apoptosis and cell cycle was detected by using flow cytometry（FCM）.Results The expression of HSV1TK mRNA was the most obvious in group L＋M＋U.The inhibition rate of proliferation induced by HSV1TK/GCV in SKOV3 was stronger than those in other groups（all P0.05）.Compared with other groups,SKOV3 cells in group L＋M＋U decreased significantly with the most abnormal morphology by light microscope.The rate of early apoptosis of SKOV3 cells in group L＋M＋U was（49.13±0.82）%,most of SKOV3 cell cycles were blocked in G1 phase（P0.05）.Conclusion Ultrasound-mediated microbubbles can enhance the anti-tumor effect of HSV1TK/GCV system by inhibiting the proliferation of SKOV3 cells and inducing the apoptosis of SKOV3 cells,and the cell cycle is blocked in G1 phase.
出处
《中国医学影像技术》
CSCD
北大核心
2012年第6期1041-1046,共6页
Chinese Journal of Medical Imaging Technology
基金
国家自然科学基金面上项目(30973169)
重庆市教委科研基金(KJ070311)
关键词
造影剂
微泡
自杀基因
卵巢肿瘤
旁观者效应
基因治疗
Contrast agents
Microbubbles
Suicide gene
Ovarian neoplasms
Bystander effect
Gene therapy
