LncRNA AC008871.1的鉴定及在肝癌中的表达

Identification of LncRNA AC008871.1 and its expression in liver cancer

目的探讨长链非编码RNA AC008871.1的鉴定及在肝癌中的表达,并预测其靶基因。方法应用USCS数据库检索其位置、CPC数据库分析其编码能力、Ensembl数据库预测其靶基因;再通过实时荧光定量PCR技术分析AC008871.1在4种肝癌细胞系(SK-Hep1、HepG2、Huh7、97L)与正常细胞系(L-02)中的差异表达,以及在肝癌组织与癌旁组织中的差异表达,并分析其与临床数据的关系。结果经数据库检索,AC008871.1位于5号染色体长臂,酪氨酸激酶FER基因3号至4号外显子之间的3号内含子的反义链上,其编码能力很弱,为非编码RNA,预测其靶基因可能为FER;经荧光定量分析,AC008871.1在不同肝癌细胞系中相对表达量均低于正常细胞,与L-02的(1.00±0.00)相比,在SK-Hep1、HepG2、Huh7、97L中的相对表达量依次是(0.32±0.22)、(0.49±0.08)、(0.83±0.03)、(0.42±0.03),差异均有统计学意义(P<0.05);肝癌组织中AC008871.1的相对表达量均低于癌旁组织[(0.29±0.21) vs1.00],差异有统计学意义(P<0.05);有吸烟史的肝癌组织中,AC008871.1的基因表达水平为(0.40±0.22),明显高于无吸烟的肝癌组织的(0.17±0.10),差异有统计学意义(P<0.05);HBEAb正常的肝癌组织中,AC008871.1的基因表达水平为(0.41±0.20),明显高于不正常的肝癌组织的(0.16±0.12),差异有统计学意义(P<0.05)。结论 AC008871.1差异表达可能与肝癌的发生、发展有密切联系,有望作为新的肝癌预后标志物和治疗靶点。

Objective To identify the long-chain non-coding RNA AC008871.1 and its expression in liver cancer, and to predict its target gene. Methods The USCS database was used to search its location, the CPC database was used to analyze its coding ability, and the Ensembl database was used to predict its target gene. Then, real-time quantitative PCR was used to evaluate the differential expression of AC008871.1 in four hepatocellular carcinoma cell lines(SKHep1, HepG2, Huh7, 97L) and in normal cell lines(L-02), as well as the differential expression in liver cancer tissues and adjacent tissues, and to analyze their relationship with clinical data. Results According to the database search,AC008871.1 is located on the antisense strand of the 3rd intron between the long arm of chromosome 5 and the exon 3to 4 of the tyrosine kinase FER gene, which is non-coding RNA and has weak coding ability, and the target gene may be predicted to be FER. By fluorescence quantitative analysis, the relative expression of AC008871.1 in different liver cancer cell lines was lower than that of normal cells: compared with L-02(1.00±0.00), the relative expression levels in SKHep1, HepG2, Huh7, and 97 L were 0.32±0.22, 0.49±0.08, 0.83±0.03, and 0.42±0.03, respectively;the difference was statistically significant(P<0.05). The relative expression of AC008871.1 in liver cancer tissues was lower than that in adjacent tissues(0.29±0.21 vs 1.00), and the difference was statistically significant(P<0.05). In the liver cancer tissues with smoking history, the gene expression level of AC008871.1(0.40±0.22) was significantly higher than that of non-smoking liver cancer tissues(0.17±0.10), and the difference was statistically significant(P<0.05). In HBEAb normal liver cancer tissues, the gene expression level of AC008871.1(0.41±0.20) was significantly higher than that of abnormal liver cancer tissues(0.16±0.12), and the difference was statistically significant(P<0.05). Conclusion The differential expression of AC008871.1 may be closely related to the occurrence and development of liver cancer, and it is expected to be a new prognostic marker and therapeutic target for liver cancer.