超声辐照联合造影微泡和盐酸阿霉素对不同卵巢癌细胞的凋亡效果评价

目的观察超声辐照联合造影微泡（SonoVue）和盐酸阿霉素（DOX）对不同卵巢癌细胞的凋亡效果,并比较同一超声波辐照剂量、超声微泡和DOX浓度杀伤两株卵巢癌细胞之间的差异。方法 MTT法检测DOX对卵巢癌A2780s细胞的毒性作用,并计算出24 h的50%抑制浓度（IC50）值。将A2780s和SKOV3两株卵巢癌细胞均分为对照组（C组）、单纯DOX组（D组）、超声＋DOX组（U＋D组）、超声＋微泡＋DOX组（U＋M＋D组）。用相同参数（超声辐照声强为0.5 W/cm2、照射时间为30 s、超声脉冲波频率为1.0 MHz）的超声波辐照,并加入相同剂量的DOX（终浓度为1.0μg/ml）及微泡（W/V为10%）,以MTT法检测两株卵巢癌细胞的各组细胞存活率;倒置显微镜观察两株卵巢癌细胞的形态学变化;并用Hoechst染色法观察A2780s卵巢癌细胞中各组细胞的凋亡情况。结果 DOX对卵巢癌A2780s细胞的IC50值约为1.0μg/ml。MTT检测发现,各处理组A2780s细胞的存活率分别为（58.2±2.5）%、（54.4±3.2）%、（52.4±1.0）%,SKOV3细胞的存活率分别为（71.4±5.9）%、（60.1±3.6）%、（58.0±4.6）%,各处理组A2780s与SKOV3细胞与各自对照组相比均有明显的增殖抑制（对照组细胞存活率均假设为100%）,但同一细胞株的各处理组间细胞存活率比较,差异无统计学意义;各处理组A2780s细胞存活率略低于对应组SKOV3细胞存活率,但差异无统计学意义;倒置显微镜观察发现各处理组A2780s与SKOV3卵巢癌细胞均出现形态学变化;Hoechst染色法进一步发现各处理组A2780s细胞核存在凋亡现象,凋亡的细胞核数量有如下趋势：对照组〈D组〈U＋D组〈U＋M＋D组,但差异无统计学意义。结论 DOX（终浓度为1.0μg/ml）体外能诱导卵巢癌细胞凋亡,但超声辐照（辐照声强为0.5 W/cm2、辐照时间为30 s、脉冲频率为1.0 MHz）与10%微泡（W/V）联合DOX并未导致该凋亡率显著增加,同时两株不同的卵巢癌

The effects of Sono Vue plus ultrasound on apoptosis of different human ovarian cancer cells induced by Doxorubicin

Objective To investigate the antitumor effect of SonoVue combined with Doxorubicin（DOX） and ultrasound on human ovarian cancer cells, and compare the differences between A2780s and SKOV3 cells treated with the same concentration of DOX and microbubble with same ultrasound parameters. Methods MTT was used to detect the growth inhibition of A2780s cells treated with DOX for 24 h. The IC50 of DOX was calculated. Both A2780s and SKOV3 cells were randomly divided into 4 groups： control group（group C）, DOX group（group D）, ultrasound ＋ DOX group（group U＋D） and ultrasound ＋ microbubble ＋ DOX group（group U＋M＋D）. The cell viability of A2780s and SKOV3 cells in each group were examined with MTT after using the same ultrasonic irradiation parameters Upon exposure to 1 MHz pulsed ultrasound beam（0.5 W/cm2） for 30 s] and the same concentration of DOX（final concentration of 1.0 μg/ml） and 10%（W/V） microbubble. The apoptotic morphology of A2780s and SKOV3 cells were observed with inverted microscope. The apoptotic nuclei morphology of A2780s cells in each group were further evaluated by Hoechst 33258 staining. Results The IC50 of DOX in A2780s cells was about 1.0 μg/ml. The survival rates of A2780s cells in DOX group, ultrasound plus DOX group and ultrasound plus microbuble combined with DOX group were（58.2±2.5）%,（54.4±3.2）% and（52.4±1.0）%, and the survival rates of SKOV3 cells were（71.4±5.9）%,（60.1±3.6）% and（58.0±4.6）%, respectively. Compared to the control group in which the control survival rate is considered as 100%, the proliferation of A2780s and SKOV3 cells in all treatment groups was significantly inhibited. However, no significant differences（P0.05） in survival rate were observed among the treatment groups of both cells. Furthermore, the survival rate of A2780s cells in each group was slightly but not statistically significant lower than that of SKOV3 cells（P 0.05）. The morphological changes of apoptosis of A2780s and SKOV3 cells were