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目的探讨不同转移能力涎腺腺样囊性癌(ACC)表达微小RNA(miRNA)的差异,以期筛选与ACC侵袭转移相关的miRNA及其所调控的靶基因。方法以ACC高转移细胞株(ACC-M)为实验组,低转移细胞株(ACC-2)为对照组,采用高通量miRNA微阵列初步筛选两株细胞中差异表达的内源性miRNA,然后挑选几个显著差异表达的miRNA,采用实时荧光定量RT-PCR(qRT-PCR)进一步加以验证,同时运用miRNA靶基因预测软件预测其可能调控的靶基因。结果芯片初筛显示,与ACC-2相比,ACC-M中表达差异具有统计学意义的miRNA分子有23个,其中上调表达的有14个(miR-15a、miR-16、miR-509-3p等),下调表达的有9个(miR-24、miR-98、miR-320等)。qRT-PCR验证与miRNAs芯片结果相符合。靶基因软件预测显示,miR-98的靶基因为Ras和高迁移率蛋白A2,miR-320的靶基因为整合素β3,miR-509-3p的靶基因为CD164,它们均与恶性肿瘤侵袭转移有关。结论不同转移能力的ACC存在差异表达的miRNA、miR-98、miR-320和miR-509-3p可能与ACC侵袭转移相关,可能通过调控相关靶基因参与ACC的侵袭转移进程。
Objective To investigate the differential expression of microRNAs (miRNAs) in different metastatic salivary adenoid cystic carcinomas (ACCs), so as to screen miRNAs involved in invasion and metastasis of ACC and its target genes. Methods ACC-2 cell line (ACC-M) was used as experimental group and ACC-2 cell line was used as control group. High-throughput miRNA microarray was used to screen differentially expressed endogenous miRNAs. Then, several miRNAs that were significantly differentially expressed were selected and further verified by real-time qRT-PCR. Meanwhile, miRNA target prediction software was used to predict the possible target genes. Results The results of microarray analysis showed that compared with ACC-2, there were 23 miRNAs with statistically significant differences in ACC-M expression among which 14 were up-regulated (miR-15a, miR-16 and miR- 3p, etc.) and down-regulated 9 (miR-24, miR-98, miR-320, etc.). qRT-PCR validation is consistent with miRNAs chip results. Prediction of target gene software showed that the target genes of miR-98 are Ras and high-mobility protein A2, the target gene of miR-320 is integrin β3, and the target gene of miR-509-3p is CD164, both of which are associated with the invasion and metastasis of malignant tumors related. Conclusions There are differentially expressed miRNAs in ACC with different metastatic potential. MiR-98, miR-320 and miR-509-3p may be involved in the invasion and metastasis of ACC, and may be involved in the invasion and metastasis of ACC by regulating related target genes.