人舌癌耐药细胞株OSC-19/CDDP的多药耐药性检测及差异性表达microRNAs的筛选

目的:检测人舌鳞癌耐药细胞株OSC-19/CDDP是否具有多药耐药的生物特性,并筛选出与亲本细胞(OSC-19)显著差异表达的microRNAs(miRNAs),探讨其在舌癌耐药中的作用。方法:对前期实验建立的耐顺铂(cisplatin,CDDP)细胞株OSC-19/CDDP,采用CCK-8法检测其对不同化疗药物的耐药性;RT-PCR检测亲代与耐药株的多药耐药基因1(multidrug resistance 1,MDR1)表达。通过微阵列芯片筛查在OSC-19/CDDP与亲本细胞OSC-19中差异性表达的miRNAs,并通过RT-PCR进一步验证基因芯片的结果。结果:人舌癌多药耐药细胞株OSC-19/CDDP对CDDP的耐药指数为16.7,交叉耐药的紫杉醇(PTX)、5-氟尿嘧啶(5-Fu)、平阳霉素(PYM)耐药指数分别为7.93、5.31和3.01;RT-PCR结果显示OSC-19/CDDP细胞中MDR1的表达是OSC-19的15.23倍。微阵列芯片结果显示:OSC-19/CDDP与OSC-19比较,分别有17个miRNAs的表达增高程度和12个miRNAs的表达降低程度超过10倍。结论:人舌鳞癌耐药细胞株OSC-19/CDDP具有多药耐药的生物学特性,可作为舌鳞癌的顺铂耐药机制及多药耐药机制研究的理想细胞模型。显著差异表达的miRNAs可能在舌鳞癌耐药的发生、发展中发挥重要作用。

Verification of the Multidrug Resistance of Human Tongue Squamous Carcinoma Cell Line OSC-19/CDDP and Its Differently Expressed MicroRNAs

Objective:To verify the multidrug resistance of OSC-19/CDDP cell line.Screening differently expressed microRNAs(miRNAs)between OSC-19/CDDP and its parental cell(OSC-19)and discussing their effect in drug resistance.Method:The drug-resistance index of cell line OSC-19/CDDP was detected to different chemotherapeutics with CCK-8,and the MDR1(multidrug resistance 1)expression in both OSC-19/CDDP and parental cell was detected with RT-PCR.Screening differently expressed miRNAs between them was detected with micro-array and confirmed RT-PCR.Results:The resistance index of OSC-19/CDDP to CDDP was 16.7 and its cross resistance index to PTX,5-FU,and PYM were 7.93,5.31 and 3.01 respectively.MDR1 mRNA expression level was 15.23 fold higher in OSC-19/CDDP than in parental cell.Micro-array revealed 17 miRNAs increased and 12 declined over 10-fold between OSC-19/CDDP and OSC-19.Conclusion:The multidrug resistance of OSC-19/CDDP was verified,making it an ideal model for mechanism research on human tongue squamous cell carcinoma cisplatin or multidrug resistance.Those differently expressed miRNAs may play vital roles in this resistance progress.