联合靶向阻断miR-29b/92b/106b抑制胃癌细胞生长及迁移

目的:本研究旨在寻找胃癌转移相关微小RNA(microRNA,miRNA,miR),揭示其对胃癌细胞生物学功能的影响,并探讨联合阻断候选miRNA在胃癌治疗中的临床应用价值。方法:收集发生淋巴结转移及无转移患者的胃癌标本各3例,采用miRNA表达芯片筛选转移相关的候选miRNA;将锁核酸修饰的候选miRNA的反义寡核苷酸转染BGC-823胃癌细胞株,运用CCK-8法、流式细胞术、划痕实验及Transwell迁移实验分析候选miRNA抑制前后胃癌细胞生物学功能的变化;构建多西环素诱导多重靶向候选miRNA的裸鼠移植瘤模型,观察联合靶向阻断候选miRNA后,肿瘤细胞体内生长情况。结果:miRNA表达芯片发现miR-29b、miR-92b及miR-106b在发生淋巴结转移患者的胃癌组织最高,并将它们确定为胃癌转移相关候选miRNA;分别在BGC-823细胞中抑制上述miRNA,可导致细胞活力减弱,凋亡诱导增加,迁移能力显著下降(P 0. 05);同时,体内联合阻断miR-29b/92b/106b,裸鼠移植瘤的形成较对照组显著减少。结论:miR-29b、miR-92b及miR-106b与胃癌细胞的迁移有关,联合阻断上述miRNA可明显抑制胃癌细胞体内外生长。这3个miRNA可能作为潜在的治疗靶点。

Combined targeting blockage of miR-29b/92b/106b inhibits gastric can-cer cell growth and migration

AIM: To investigate the metastasis-associated microRNAs (miRNAs, miR) in gastric cancer, and to determine their biological and therapeutic roles in this malignancy. METHODS: The tumor tissue samples from gastric cancer patients with or without lymph node metastasis were collected (n=3 each). miRNA microarray was used to determine the metastasis-associated miRNAs. Gastic cancer cell line BGC-823 was transfected with locked nucleic acidmodified antisense oligonucleotides of candidate miRNAs and subsequently used for functional assays including CCK-8 assay, flow cytometry, wound healing assay and Transwell migration assay. Furthermore, the in vivo xenograft mice were used to evaluate the tumor suppressive effect of collaborative inhibition of the candidate miRNAs. RESULTS: miR-29b, miR-92b and miR-106b were up-regulated in the tumor tissues from the gastric cancer patients with lymph node metastasis. The functional assays showed that blockage of miR-29b, miR-92b and miR-106b by antisense oligonucleotides in the BGC-823 cells significantly inhibited cell growth and migration, and induced apoptosis. Furthermore, the collaborative inhibition of these triple miRNAs remarkably suppressed tumor growth in vivo. CONCLUSION: miR-29b, miR-92b and miR-106b are metastasis-associated miRNAs. These miRNAs may provide promising therapeutic targets in gastric cancer.