Gao Ya-Wen, Ma Fang, Xie Yang-Chun, Ding Meng-Ge, Luo Li-Hua, Jiang Shun, Rao Le, Liu Xian-Ling
Department of Oncology, The Second Xiangya Hospital of Central South University Changsha 410011, Hunan, China.
Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430022, Hubei, China.
Am J Transl Res. 2019 Aug 15;11(8):4761-4775. eCollection 2019.
Long non-coding RNA tissue differentiation-inducing non-protein coding (TINCR) is associated with the carcinogenesis of several cancers. However, little is known about the function and mechanism of TINCR in lung adenocarcinoma (LUAD). Here, we aimed to analyze expression of TINCR and elucidate its mechanistic involvement in the progression of LUAD. The expression of TINCR was investigated according to Gene Expression Profiling Interactive Analysis at first and then detected in 29 LUAD tissues and paired adjacent normal tissues using qRT-PCR. Results indicated that TINCR was evidently downregulated in LUAD. The association between TINCR and clinicopathological parameters was analyzed by Pearson's chi-square test, suggesting TINCR was closely correlated with TNM stage and lymph mode metastasis. Subsequently, the function role of TINCR was examined by gain- and loss-of-function studies in LUAD (A549 and NCI-H292) cells. As analyzed by the scratch wound-healing and transwell assays, results revealed that TINCR suppressed the migration and invasion of A549 and NCI-H292 cells. However, TINCR exerted no effects on the cell proliferation as determined by CCK8 assay. Furthermore, we reported that loss of Sp1 could inhibit TINCR expression. Expressions of miR-107/miR-1286 were detected by qRT-PCR assay in A549 and NCI-H292 cells after TINCR knockdown or overexpression. In addition, the direct binding ability of the predicted miR-107 or miR-1286 binding site on TINCR was validated by luciferase activity assay. Results indicated TINCR could constrain the expression of miR-107/miR-1286, and was a target of them in LUAD cells. Bioinformatics analyses showed that BTRC and RAB14 was the potential target gene of miR-107 and miR-1286, respectively. These data revealed a possible regulatory mechanism in which upregulation of TINCR induced by Sp1 could constrain the migration and invasion through regulating miR-107 or miR-1286 in LUAD cells. Conjointly, our findings provide a valuable insight into the regulatory mechanism of TINCR in LUAD, supportive to its potential of therapeutic target for LUAD patients.
长链非编码RNA组织分化诱导非蛋白质编码基因（TINCR）与多种癌症的发生相关。然而，关于TINCR在肺腺癌（LUAD）中的功能和机制知之甚少。在此，我们旨在分析TINCR的表达，并阐明其在LUAD进展中的作用机制。首先根据基因表达谱交互分析研究TINCR的表达，然后使用qRT-PCR在29例LUAD组织及配对的癌旁正常组织中进行检测。结果表明，TINCR在LUAD中明显下调。通过Pearson卡方检验分析TINCR与临床病理参数之间的关联，提示TINCR与TNM分期和淋巴结转移密切相关。随后，通过在LUAD（A549和NCI-H292）细胞中进行功能获得和缺失研究来检测TINCR的功能作用。通过划痕伤口愈合和Transwell实验分析，结果显示TINCR抑制A549和NCI-H292细胞的迁移和侵袭。然而，CCK8实验表明TINCR对细胞增殖没有影响。此外，我们报道Sp1的缺失可抑制TINCR表达。在TINCR敲低或过表达后，通过qRT-PCR实验检测A549和NCI-H292细胞中miR-107/miR-1286的表达。此外，通过荧光素酶活性实验验证了TINCR上预测的miR-107或miR-1286结合位点的直接结合能力。结果表明TINCR可抑制miR-107/miR-1286的表达，并且在LUAD细胞中是它们的靶标。生物信息学分析表明，BTRC和RAB14分别是miR-107和miR-1286的潜在靶基因。这些数据揭示了一种可能的调控机制，即Sp1诱导的TINCR上调可通过调节LUAD细胞中的miR-107或miR-1286来抑制迁移和侵袭。总之，我们的研究结果为TINCR在LUAD中的调控机制提供了有价值的见解，支持其作为LUAD患者治疗靶点的潜力。
