长链非编码RNA AFAP1-AS1促进结肠癌细胞侵袭的作用和机制

The role and mechanism of long non-coding RNA AFAP1-AS1 in promoting colon cancer cells invasion

目的探讨长链非编码RNA AFAP1-AS1对结肠癌细胞侵袭的调控作用及分子机制。方法采用实时定量PCR的方法检测56例结肠癌组织和36例癌旁组织中AFAP1-AS1的表达水平,分析AFAP1-AS1表达与结肠癌患者临床病理特征及预后的相关性;通过腺病毒感染的方法在结肠癌细胞系中过表达AFAP1-AS1,细胞穿孔实验确定AFAP1-AS1对结肠癌细胞侵袭能力的影响,通过实时定量PCR和蛋白质免疫印迹的方法检测AFAP1-AS1对AFAP1表达的影响。结果结肠癌组织中AFAP1-AS1表达明显高于癌旁组织(P<0.05),其高表达与患者年龄、性别和肿瘤分化均无相关性(均P>0.05),而与淋巴结转移、临床分期相关(均P<0.05);AFAP1-AS1高表达患者的总生存期和无瘤生存率较低表达患者明显降低,差异有统计学意义(P<0.05);感染腺病毒后,结肠癌细胞AFAP1-AS1水平明显增加,穿膜细胞数目也明显增多(P<0.05);AFAP1-AS1过表达能抑制AFAP1mRNA和蛋白的表达,信息学预测发现AFAP1-AS1序列上存在3个AFAP1的互补结合区域。结论 AFAP1-AS1在结肠癌组织高表达,可能通过抑制AFAP1表达促进结肠癌细胞侵袭能力,提示AFAP1-AS1可能是结肠癌诊断治疗和预后的潜在靶点和重要指标。

Objectlve To investigate the role and molecular mechanism of long non-coding RNA AFAP1-AS1 in colon cancer cells invasion. Methods The expression level of AFAP1-AS1 in colon cancer tissues （n=56） and ad- jacent tissues （n=36） was detected by Real-time PCR assay, and the relationship between AFAP1-AS1 levels and cli- nical pathological features and prognosis was evaluated. AFAP1-AS1 overexpression was implemented by adenovi- rus, and the invasion ability of colon cancer cells was determined by trans-well assay. Real-time PCR and western blot assays were applied to confirm the effect ofAFAP1-AS 1 on AFAP1 expression. Results The expression of AFAP1- AS 1 in colon cancer tissues was significantly higher than that in the adjacent tissues （19 〈 0.05）. The high expression of AFAP 1-AS 1 was significantly correlated with lymph node metastasis and clinical stage （allP 〈 0.05）, while it was not correlated with age, gender and tumor differentiation （allP 〉 0.05）. The overall survival and disease-free survival in colon cancer patients with high expression of AFAP1-AS1 were significantly lower than those in colon cancer pa- tients with low expression ofAFAP1-AS1 （allP 〈 0.05）. After adenovirus infection, the expression of AFAP1-AS1 in colon cancer cells were significantly increased, and the number oftransmembrane cells were also significantly in- creased （all P 〈 0.05）. The mRNA and protein expression of AFAP1 was significantly inhibited by overexpression of AFAP1-AS1. Bioinformatics prediction showed that there were three complementary regions in AFAP1-AS1 to bind with AFAP1 mRNA. Conclusions AFAP1-AS1 was highly expressed in colon cancer and could promote the invasion ability of colon cancer cells by inhibiting AFAP 1 expression, suggesting that AFAP 1-AS 1 might be a poten- tial target and important index for diagnosis and prognosis of colon cancer.