超声微泡介导自杀基因对卵巢癌细胞的抗瘤效应

摘要

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 plas-mid 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), li-posome 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 HSV1 TK 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 apop-tosis and cell cycle was detected by using flow cytometry (FCM). Results The expression of HSV1 TK 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 P<0. 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 (P<0. 05). Conclusion Ultrasound-mediated microbubbles can enhance the anti-tumor effect of HSV1 TK/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.%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-HSV1 TK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HSV1 TK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.