RAB5A反义寡脱氧核苷酸抑制大肠癌细胞转移侵袭的实验研究

RAB5A antisense oligonucleotides inhibit infiltration and metastasis of colorectal cancer

目的应用RAB5A反义寡脱氧核苷酸(ASODN)体外转染高转移度的大肠癌细胞LS174T,观察其对肿瘤细胞的增殖、侵袭和运动的影响,初步探讨RAB5A在大肠癌转移中的作用与机制。方法用脂质体将RAB5A转染人大肠癌高转移细胞LS174T,细胞增殖抑制实验(MTT法)检测RAB5A反义寡核苷酸对细胞的生长和增殖的影响;Transwell小室法检测对细胞的侵袭和运动能力的影响,明胶酶谱法检测细胞基质金属蛋白酶2(MMP2)的分泌与活性的变化;RT-PCR法检测大肠癌细胞的RAB5A的mRNA表达。结果RAB5A-ASODN能抑制大肠癌高转移细胞LS174T的增殖和高转移大肠癌细胞的浸润和运动能力,并能抑制基质金属蛋白酶的分泌和活性。RAB5A-ASODN组RAB5A表达量比对照组明显减少。结论在体外抑制RAB5A的表达可抑制大肠癌的转移。

Objective To explore the effects and mechanism of RAB5A antisense oligonucleotides （RAB5A-ASODN） in colorectal cancer metastasis. Methods RAB5A-ASODN were transferred into human colorectal cancer cell lines LS174T. MTT assay was used to detect the effects of RAB5A antisense oligonucleotides on LS174T cell muhiplification. Transwell cabin was used to evaluate the changes of activities of adhesion, invasion, motion. Gelatinase assay was used to examine the secretion and activity change of matrix metalloprotease MMP2. RT-PCR assay was used to test RAB5A mRNA expression. Results MTI＇ results showed that RAB5A- ASODN could inhibit the proliferation of high metastasis colorectal cell line LS174T. RAB5A-ASODN could al- so inhibit the invasion and motion activities of LS174T cells. Gelatinase assay showed that RAB5A-ASODN can inhibit the secretion and activity of MMP2. Conclusion Metastasis of colorectal cancer can be suppressed when RAB5A expression is inhibited by RAB5A-ASODN.