长链非编码RNA LINC00483抑制miR-204-3p对结直肠癌细胞侵袭转移的实验研究

Experimental study of the inhibitory effects of Long non-coding RNA LINC00483 on miR-204-3p expression promoting colorectal cancer cells metastasis

目的探讨长链非编码RNA LINC00483能否通过抑制miR-204-3p表达的方式促进结直肠癌细胞侵袭转移。方法qRT-PCR法检测长链非编码RNA LINC00483及miR-204-3p在结直肠癌组织及细胞系HT29和LOVO中的表达;考察不同LINC00483及miR-204-3p表达水平分别与结直肠癌患者临床病理参数的相关性;在HT29及LOVO中分别转染特异性LINC00483干扰寡核苷酸(siLINC00483)以下调其表达;qRT-PCR法检测下调LINC00483后HT29及LOVO细胞内miR-204-3p表达变化,Transwell实验检测HT29及LOVO细胞侵袭转移能力的变化;共转染siLINC00483及miR-204-3p inhibitor,Transwell实验再次检测HT29及LOVO细胞侵袭转移能力的变化。结果相比于癌旁组织和正常肠上皮细胞系NCM460,LINC00483在结直肠癌组织及细胞系中表达增高而miR-204-3p表达降低(P<0.01);高表达的LINC00483和低表达的miR-204-3p均与结直肠癌患者的不良病理参数密切相关(P<0.01);下调LINC00483可明显抑制HT29和LOVO细胞中miR-204-3p的表达并抑制其侵袭转移(P<0.01);相比于单独转染siLINC00483,共转染siLINC00483及miR-204-3p inhibitor后,HT29及LOVO细胞的侵袭转移能力明显增加(P<0.01)。结论LINC00483在结直肠癌中表达增高且可通过抑制miR-204-3p的方式促进结直肠癌细胞侵袭转移。

Objective To investigate whether LINC00483 could promote colorectal cancer cells metastasis via suppressing of miR-204-3p expression.Methods A qRT-PCR assay was used to detect the expression of long noncoding RNA LINC00483 and miR-204-3p in colorectal cancer(CRC)tissue specimens and in CRC cell lines HT29 and LOVO.The correlation between LINC00483 or mIR-204-3p and clinicopathological features of patients with CRC was analyzed by means of Pearson Chi-Square test.Specific small interfering RNAs of LINC00483(siLINC00483)was transfected into HT29 and LOVO cells to knockdown the expression of LINC00483.A qRT-PCR assay was performed to measure the changes of miR-204-3p in HT29 and LOVO cells.A transwell assay was used to check the metastatic ability of HT29 and LOVO cells.After a cotransfection of siLINC00483 and miR-204-3p inhibitor,the metastatic changes of HT29 and LOVO were detected by Transwell assay.Results As compared with para-tumor tissue and normal colon epithelial cell line NCM460,the expression levels of LINC00483 were increased in CRC tissues and cell lines,however,the expression levels of miR-204-3p were decreased.The high expression of LINC00483 and low expression of miR-204-3p were closely correlated with poor clinicopathological features of patients with CRC.Downregulation of LINC00483 could remarkably suppress miR-204-3p expression and inhibit the metastasis in HT29 and LOVO cells.Conclusion The expression levels of LINC00483 are increased in CRC tissues,which can promote colorectal cancer cells metastasis via suppressing of miR-204-3p expression.