摘要
目的检测miR-196a在人胃癌组织及细胞系中的表达,探讨抑制或过表达miR-196a对胃癌细胞侵袭转移能力的影响,以及其可能作用的靶基因。方法通过实时定量PCR技术检测胃癌组织及细胞系中miR-196a的表达水平,通过转染miR-196a inhibitors或mimics抑制或上调其表达,并通过定量PCR检测转染效率。利用划痕迁移实验、Transwell侵袭实验和MTT实验检测上调或下调miR-196a水平对MGC-803细胞的迁移、侵袭和增殖能力的影响。采用生物信息学及Western blotting方法验证miR-196a对靶基因HOXA5的调控机制。结果相对于正常胃黏膜组织及细胞,胃癌组织和细胞系中miR-196a的表达水平显著上调(上调约28倍,P<0.01),MGC-803细胞中转染miR-196a inhibitors或mimics能显著抑制(下降了53%,P<0.01)或上调(上调约8倍,P<0.01)miR-196a表达水平。抑制miR-196a表达能降低MGC-803细胞的迁移、侵袭和增殖能力,而上调其表达则相反。miR-196a能够负性调控HOXA5的表达。结论胃癌组织及细胞系中miR-196a的表达上调可能通过抑制HOXA5的表达显著提高胃癌细胞的侵袭转移能力,促进胃癌的发生、发展。
Objective To investigate the expression level of miR-196a in gastric cancer（ GC） tissues and cell lines, to study the effect of inhibition or up-regulation of miR-196a expression on GC cells migration and invasion, and to find out the possible target gene. Methods Real-time quantitative PCR was performed to detect the relative expression of miR-196a in GC cell lines and tissues. MiR-196a inhibitors or mimics were transfected into MGC-803 cells to down-regulate or up-regulate miR-196a expression, and Real-time quantitative PCR was used to test the transfection efficiency. Wound healing, transwell and MTT assays were performed to detect the effect of miR-196a on GC cells migration, invasion and proliferation. Bioinformatics and Western blotting analysis were performed to investigate the target genes that could be regulated by miR-196a. Results This study showed that miR-196a was highly expressed both in GC samples and cell lines compared with their corresponding normal tissues and cell lines （approximately 28 folds, P＆lt;0？01）. Transfection of miR-196a inhibitors or mimics could significantly decrease （ 53%, P＆lt;0？01） or increase （ approximately 8 folds, P＆lt;0？01） its expression in MGC-803 cells, respectively. And down-regulation of miR-196a expression could inhibit MGC-803 cells migra-tion, invasion and proliferation by down-regulating HOXA5 expression. Conclusion Up-regulated miR-196a could promote gastric cancer cells migration and invasion by inhibiting the expression of HOXA5, which may affect the development of gastric cancer.
出处
《临床肿瘤学杂志》
CAS
2014年第3期193-198,共6页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(81070620)
关键词
胃癌
迁移
侵袭
HOXA5
miRNA
miR-196a
Gastric cancer
miRNA
miR-196a
Migration
Invasion
