利用外泌体评估卵巢癌患者对顺铂敏感性

目的:利用外泌体评估卵巢癌患者对顺铂敏感性。方法:超速离心法从卵巢癌细胞A2780、SKOV3、CAOV3培养上清及卵巢癌患者血清中提取外泌体。MTT法检测顺铂对卵巢癌细胞株的24h时半数抑制浓度(IC50),检测出顺铂敏感细胞及耐药细胞。MTT法检测敏感细胞A2780在细胞外泌体与顺铂共培养条件下的增殖。Caspase 3/7活性凋亡检测A2780细胞在细胞外泌体与顺铂共培养条件下的凋亡。MTT法检测A2780细胞在卵巢癌患者血清外泌体与顺铂共培养条件下的增殖,并随访卵巢癌患者病情进展情况。结果:通过超速离心法能从卵巢癌细胞系培养上清及卵巢癌患者外周血清中分离出外泌体。MTT分析证实A2780为顺铂敏感细胞,SKOV3、CAOV3为顺铂不敏感细胞。细胞系来源的外泌体能显著降低A2780对顺铂的敏感性(P<0.05),同时削弱顺铂对A2780中Caspase 3/7的活性(P<0.05),其中SKOV3和CAOV3外泌体的抑制作用显著高于A2780。基于A2780细胞+顺铂模型,将30例卵巢癌患者分为顺铂敏感组和不敏感组,其中不敏感组的13例患者的血清外泌体降低A2780对顺铂敏感性(P<0.05),术后1年已有5例患者有复发情况,而敏感组无复发。结论:利用A2780细胞模型分析外泌体对顺铂杀伤作用的影响,能在一定程度上评估卵巢癌患者对顺铂的敏感性。

Utilization of exosomes in evaluating cisplatin sensitivity of patients with ovarian cancer

Objective:To evaluate cisplatin sensitivity of ovarian cancer patients by using exosomes.Methods:Exosomes were extracted from the ovarian cancer cell A2780,SKOV3,CAOV3 culture supernatant and ovarian cancer patient serum by ultracentrifugation.The 24-hour half-inhibitory concentration(IC50) of cisplatin against each ovarian cancer cell line was detected by MTT assay,and cisplatin-sensitive cells and resistant cells were detected.MTT assay was used to detect the proliferation of sensitive cells A2780 under co-culture conditions of exosomes and cisplatin.Caspase 3/7 activity was tested for apoptosis in A2780 under co-culture conditions of exosumes and cisplatin.MTT assay was used to detect the proliferation of A2780 in co-culture with serum exosomes and cisplatin from ovarian cancer patients,and progression of ovarian cancer patients was followed up.Results:Exosomes could be isolated from the culture supernatant of ovarian cancer cells and the serum of ovarian cancer patients by ultracentrifugation.MTT analysis confirmed that A2780 was a cisplatin-sensitive cell,and SKOV3 and CAOV3 were cisplatin-insensitive cells.Cell-derived exosomes significantly reduced the sensitivity of A2780 to cisplatin(P<0.05),and attenuated the activation of caspase 3/7 in A2780 by cisplatin(P<0.05),in which the inhibition of SKOV3 and CAOV3 exosomes was significant higher than A2780.Based on the A2780 cell + cisplatin model,30 patients with ovarian cancer were divided into cisplatin-sensitive and insensitive groups,and exosomes of 13 patients decreased A2780 sensitivity to cisplatin(P<0.05).Five patients had recurrence 1 year after surgery,while the sensitive group had no recurrence.Conclusion:The effect of exosomes from patients on the cisplatin sensitivity of A2780 could be utilized as a model to evaluate the cisplatin treatment result on ovarian cancer patients.