长链非编码 RNA SNHG7 通过调控 miR-449a/TGIF2 轴促进非小细胞肺癌细胞增殖、迁移、侵袭及上皮间质转化。
Long noncoding RNA SNHG7 contributes to cell proliferation, migration, invasion and epithelial to mesenchymal transition in non-small cell lung cancer by regulating miR-449a/TGIF2 axis.
机构信息
Department of Respiratory Medicine, Yantai Yuhuangding Hospital, Yantai, China.
Department of Respiratory Medicine, Yantai Muping District Traditional Chinese Medical Hospital, Yantai, China.
出版信息
Thorac Cancer. 2020 Feb;11(2):264-276. doi: 10.1111/1759-7714.13245. Epub 2019 Dec 3.
BACKGROUND
Non-small cell lung cancer (NSCLC) is an intractable malignant lung cancer with high rates of metastasis and mortality. Currently, long noncoding RNA nuclear RNA host gene 7 (SNHG7) is recognized as a biomarker of multiple cancers. However, the role of SNHG7 in NSCLC requires further understanding.
METHODS
The expression of SNHG7, miR-449a and TGIF2 in NSCLC tumors and cells was examined by quantitative real time polymerase chain reaction (qRT-PCR). Cell viability was measured by MTT assay. Cell migration and invasion was conducted using transwell assay. Protein expression of TGIF2, vimentin, N-cadherin and E-cadherin was detected by western blot. The interaction between miR-449a and SNHG7 or TGIF2 was determined by luciferase reporter system, RIP and RNA pull-down assay, respectively. Xenograft mice models were established by subcutaneously injecting A549 cells transfected with sh-SNHG7 and sh-control.
RESULTS
SNHG7 expression was upregulated in NSCLC tumors and cells compared with normal tissues and cells. SNHG7 silencing repressed cell proliferation, migration, invasion and epithelial to mesenchymal transition (EMT) in NSCLC. Consistently, SNHG7 knockdown hindered tumor growth in vivo. The subsequent luciferase reporter system, RIP and RNA pull-down assay validated the interaction between miR-449a and SNHG7 or TGIF2. The rescue experiments displayed that miR-449a inhibitor counteracted SNHG7 silencing induced inhibition on proliferation, migration, invasion and EMT. Similarly, restoration of TGIF2 reversed miR-449a mediated inhibition on cell progression. In addition, the results indicated that SNHG7 could regulate cell progression by targeting miR-449a/TGIF2 axis.
CONCLUSION
SNHG7 contributed to cell proliferation, migration, invasion and EMT in NSCLC by upregulating TGIF2 via sponging miR-449a, representing a novel targeted therapy method for NSCLC.
背景
非小细胞肺癌（NSCLC）是一种难以治疗的恶性肺癌，其转移率和死亡率都很高。目前，长链非编码 RNA 核 RNA 宿主基因 7（SNHG7）被认为是多种癌症的生物标志物。然而，SNHG7 在 NSCLC 中的作用需要进一步了解。
方法
通过实时定量聚合酶链反应（qRT-PCR）检测 NSCLC 肿瘤和细胞中 SNHG7、miR-449a 和 TGIF2 的表达。通过 MTT 测定法测量细胞活力。使用 Transwell 测定法进行细胞迁移和侵袭实验。通过 Western blot 检测 TGIF2、波形蛋白、N-钙粘蛋白和 E-钙粘蛋白的蛋白表达。通过荧光素酶报告系统、RIP 和 RNA 下拉实验分别确定 miR-449a 与 SNHG7 或 TGIF2 的相互作用。通过皮下注射转染 sh-SNHG7 和 sh-control 的 A549 细胞建立异种移植小鼠模型。
结果
与正常组织和细胞相比，SNHG7 在 NSCLC 肿瘤和细胞中表达上调。SNHG7 沉默抑制 NSCLC 细胞的增殖、迁移、侵袭和上皮间质转化（EMT）。同样，SNHG7 敲低抑制体内肿瘤生长。随后的荧光素酶报告系统、RIP 和 RNA 下拉实验验证了 miR-449a 与 SNHG7 或 TGIF2 的相互作用。挽救实验表明，miR-449a 抑制剂抵消了 SNHG7 沉默对增殖、迁移、侵袭和 EMT 的抑制作用。同样，恢复 TGIF2 逆转了 miR-449a 对细胞进展的抑制作用。此外，结果表明，SNHG7 通过靶向 miR-449a/TGIF2 轴调节细胞进展。
结论
SNHG7 通过上调 TGIF2 来促进 NSCLC 细胞的增殖、迁移、侵袭和 EMT，通过海绵吸附 miR-449a，为 NSCLC 提供了一种新的靶向治疗方法。
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