Department of Thoracic Surgery, Affiliated Hospital, Qingdao University, Qingdao, China.
Department of Infectious Disease, Affiliated Hospital, Qingdao University, Qingdao, China.
Thorac Cancer. 2018 Aug;9(8):939-949. doi: 10.1111/1759-7714.12767. Epub 2018 Jun 13.
The antisense of the OIP5-AS1 gene is a long non-coding RNA (lncRNA) that is reported to be upregulated and promotes cell proliferation in multiple human cancers; however, its function in lung cancer is unknown. We investigated the regulatory function and underlying mechanisms of OIP5-AS1 in lung cancer.
OIP5-AS1 and microRNA (miR)-378a-3p expression were assayed by quantitative real-time PCR, and proliferation-related protein expression was measured by Western blotting. Cell viability was detected using methyl thiazolyl tetrazolium assay. Luciferase reporter assay and RNA immunoprecipitation were used to detect the direct regulation of miR-378a-3p by OIP5-AS1. Nude mice were used to test the function of OIP5-AS1 in vivo.
OIP5-AS1 was highly expressed in lung cancer tissues and was correlated with tumor size and tumor growth speed. OIP5-AS1 overexpression increased lung cancer cell proliferation in vitro. Further investigation revealed that OIP5-AS1 functions as a competing endogenous RNA of miR-378a-3p. MiR-378a-3p overexpression inhibited cell proliferation and caused proliferation-associated proteins CDK4 and CDK6 to decrease in A549 cells. Overexpression of wild type OIP5-AS1 led to strong CDK4 and CDK6 expression; however, these two proteins did not change when mutated OIP5-AS1 was upregulated. Finally, in vivo assay showed that the speed of tumor growth was increased and decreased when OIP5-AS1 was upregulated and downregulated, respectively.
Our results revealed that OIP5-AS1 acts as a growth-promoting lncRNA in lung cancer by suppressing miR-378a-3p function. OIP5-AS1 and miR-378a-3p interaction may provide a potential target for lung cancer treatment.
OIP5-AS1 基因的反义是一种长链非编码 RNA（lncRNA），据报道在多种人类癌症中上调并促进细胞增殖；然而，其在肺癌中的功能尚不清楚。我们研究了 OIP5-AS1 在肺癌中的调节功能和潜在机制。
通过实时定量 PCR 测定 OIP5-AS1 和 microRNA（miR）-378a-3p 的表达，通过 Western blot 测定增殖相关蛋白的表达。使用甲基噻唑基四唑测定法检测细胞活力。使用荧光素酶报告基因测定和 RNA 免疫沉淀检测 miR-378a-3p 对 OIP5-AS1 的直接调节。使用裸鼠检测 OIP5-AS1 在体内的功能。
OIP5-AS1 在肺癌组织中高表达，与肿瘤大小和肿瘤生长速度相关。OIP5-AS1 过表达增加体外肺癌细胞增殖。进一步研究表明，OIP5-AS1 作为 miR-378a-3p 的竞争内源性 RNA 发挥作用。miR-378a-3p 过表达抑制细胞增殖并导致 A549 细胞中增殖相关蛋白 CDK4 和 CDK6 减少。野生型 OIP5-AS1 的过表达导致 CDK4 和 CDK6 表达强烈增加；然而，当突变型 OIP5-AS1 上调时，这两种蛋白质没有变化。最后，体内实验表明，当 OIP5-AS1 上调和下调时，肿瘤生长速度分别增加和减少。
我们的结果表明，OIP5-AS1 通过抑制 miR-378a-3p 功能在肺癌中充当促生长 lncRNA。OIP5-AS1 和 miR-378a-3p 相互作用可能为肺癌治疗提供潜在靶点。
