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本研究观察低频低功率超声联合阿霉素(adriamycin,A02)对白血病耐药细胞株K562/A02的影响,寻找合适的干预参数,探讨耐药细胞耐药逆转的可能机制。取对数生长期的白血病K562/A02细胞株,将研究对象分为空白组、单纯阿霉素组(A02)、单纯超声组(US)、阿霉素加超声组(A02+US)共4组。细胞在24孔培养板上受低频低功率超声作用,用台盼蓝拒染法、MTT法检测细胞活性,应用瑞氏染色法和流式细胞术分别检测细胞凋亡和药物浓度,扫描电子显微镜观察细胞表面变化。结果显示,频率20 kHz、声强0.25 W/cm2、作用时间60秒的超声,可显著降低阿霉素的LC50;当声强0.50 W/cm2、作用时间大于30秒的超声,对细胞有急性杀伤作用。细胞经低频超声作用后细胞膜表面出现直径大约1-2μm的小孔,细胞内阿霉素的药物浓度增加,细胞凋亡增强。结论:适当参数的超声辐射通过超声空化导致肿瘤细胞产生声孔效应,使阿霉素对耐药细胞株的抑制作用增强,从而逆转耐药细胞株的耐药性。
This study was to observe the effect of low frequency low power ultrasound combined with adriamycin (A02) on the leukemia cell line K562 / A02, find suitable intervention parameters and explore the possible mechanism of drug resistance reversal. The logarithmic growth phase leukemia K562 / A02 cell lines were divided into four groups: blank group, A02 group, US group, A02 + US group group. The cells were exposed to low frequency and low power ultrasound on 24-well plates. Cell viability was detected by MTT assay with trypan blue exclusion method. Cell apoptosis and drug concentration were detected by Wright staining and flow cytometry, respectively. Scanning electron microscopy Observation of cell surface changes. The results showed that the frequency of 20 kHz, the sound intensity of 0.25 W / cm2, the action time of 60 s ultrasound, can significantly reduce the doxorubicin LC50; when the sound intensity of 0.50 W / cm2, the role of time greater than 30 seconds of ultrasound, acute cells Killing effect. After the cells were treated by low frequency ultrasound, small holes with a diameter of about 1-2μm appeared on the surface of the cell membrane. The concentration of doxorubicin in the cells increased and the apoptosis of the cells increased. Conclusion: Ultrasound radiation with appropriate parameters can induce sonic hole in tumor cells by ultrasonic cavitation, and enhance the inhibitory effect of doxorubicin on drug-resistant cell lines, thus reversing the drug resistance of drug-resistant cell lines.