| 摘要: |
| 目的 探讨超声辅助卡铂抑制体外A549细胞活性的适宜卡铂剂量及超声能量参数。方法 96孔板接种A549细胞,加入浓度梯度0-400>μg/ml的卡铂培养24小时后,每孔加入 10μlCCK8 溶液,孵育 1.5 小时,酶标仪测定在450nm处的吸光度值,计算细胞抑制率,应用Graphpad Prism 5软件的“药物剂量-效应模型”计算卡铂24小时IC50。设置空白组、US组、USMB组、CBP组、CBP+US组、CBP+USMB组,卡铂浓度为50 μg/ml,声强梯度范围为0.2-2.2W/cm2,频率1MHz,占空比10%,辐照时间1分钟,培养24小时,CCK8法测细胞存活率,应用SPSS24.0分析存活率差异,筛选有效剂量。光学倒置显微镜观察各组细胞形态学变化。结果 卡铂24小时IC50为65.72μg/ml。抑制A549细胞活性的强弱排序为CBP+USMB组>CBP+US组>CBP组>USMB组>US组,CBP+USMB组与其余4组之间差异有统计学意义(P<0.001),CBP(50μg/ml)+USMB(0.2-1.0W/cm2)5组之间差异无统计学意义(P>0.05)。处理组均可见细胞形态不规则、贴壁性降低,以CBP+USMB组为著。结论 卡铂50μg/ml、声强0.6W/cm2为24小时联合疗法适宜剂量。超声联合微泡可以与卡铂协同抑制A549细胞活性,成为肿瘤联合疗法的新策略。 |
| 关键词: 超声空化 超声联合微泡 肿瘤 卡铂 药物递送 |
| DOI： |
| 投稿时间：2018-05-04修订日期：2018-06-12 |
| 基金项目:国家自然科学基金(81371553) |
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| Study on the Appropriate Dose of Ultrasound Combined with Microbubble Assisting Carboplatin in Inhibiting A549 Cell line Activity |
| ZHUO Zhongxiong,LI Yue |
| (Department of Ultrasound,202 Hospital of PLA;Department of Ultrasound，the Second Affiliated Hospital of Army Medical University) |
| Abstract: |
| Objective To investigate the appropriate dose of carboplatin and ultrasonic energy parameters for ultrasound assisting carboplatin on A549 cell line in vitro. Methods A549 cells were plated in 96-well plates, and carboplatin was added in a concentration gradient of 0-400μg/ml for 24 hours. Then 10μl CCK-8 was added to each well and cells were incubated for 1.5 hours. The absorbance at 450 nm was measured to calculate the cell inhibition rate. The IC50 of carboplatin for 24-hour was calculated using the Graphpad Prism 5 software with drug dose-effect model. Blank group, US group, USMB group, CBP group, CBP+US group, CBP+USMB group were set under condition of 50 μg/ml carboplatin, low intensity gradient range of untrasound from 0.2 to 2.2W/cm2, US frequency =1MHz; duty cycle =10%, sonication time=1 minute. After 24 hours, cell viability was measured by CCK8 assay. Differences of survival rates were analyzed using SPSS24.0, and appropriate dose range was discovered. Optical inverted microscope was used to observe the morphological changes of cells in each group. Results The IC50 of carboplatin for 24 hours was 65.72μg/ml. The inhibitory effect was ranked as CBP+USMB group>CBP+US group>CBP group>USMB group>US group, the difference was significant (P<0.001). However, no significant difference was found among the five CBP(50μg/ml)+USMB(0.2-1.0W/cm2) groups(P>0.05). In the treatment groups, some cells were in irregular shape and adherence was reduced, which was most significant in the CBP+USMB group. Conclusion 50μg/ml carboplatin and intensity 0.6W/cm2 are the appropriate dose for CBP+USMB therapy for A549 Cell line in vitro. Ultrasound combined with microbubbles can assist carboplatin and inhibit the viability of A549 cells. It may become a new strategy for cancer therapy. |
| Key words: Ultrasound cavitation Ultrasound combined with microbubbles Tumors Carboplatin Drug delivery |
