| 李 凌,陈思翰,朱德东.Linc00176靶向miR-338-3/SMO轴对肝癌细胞增殖和迁移的影响[J].肿瘤学杂志,2019,25(10):858-863. |
| Linc00176靶向miR-338-3/SMO轴对肝癌细胞增殖和迁移的影响 |
| Effect of Linc00176 Targeting MiR-338-3p/SMO Axis on Proliferation and Migration of Hepatoma Cells |
| 投稿时间：2019-05-16 |
| DOI：10.11735/j.issn.1671-170X.2019.10.B002 |
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| 中文关键词: Linc00176 肝肿瘤 miR-338-3p SMO 增殖 迁移 |
| 英文关键词:Linc00176 hepatocellular carcinoma miR-338-3p SMO proliferation migration |
| 基金项目: |
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| 摘要点击次数: 1296 |
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| 中文摘要: |
| 摘 要：［目的］ 探讨基因间长链非编码RNA Linc00176靶向miR-338-3p/SMO对肝癌（HCC）细胞增殖和迁移的影响。［方法］ 收集手术切除的肝癌患者的HCC组织和癌旁组织标本各96例，采用荧光定量PCR检测Linc00176和miR-338-3p在HCC组织和癌旁组织中的表达水平；构建shRNA-Control、 shRNA-Linc00176、miRNA-Control和miR-338-3p慢病毒肝癌细胞系，采用生物信息学和双荧光素酶报告基因检测分析Linc00176和miR-338-3p的靶向关系；采用CCK-8法和异种移植瘤实验检测各组细胞的增殖能力；采用Transwell分析各组细胞的迁移能力；采用Western blot检测各组细胞中SMO蛋白的表达水平。［结果］ Linc00176在HCC组织中的mRNA相对表达水平（1.94±0.21）显著高于癌旁组织（0.57±0.10）；miR-338-3p在HCC组织中的mRNA相对表达水平（0.35±0.08）显著低于癌旁组织（1.62±0.15），差异均有统计学意义（P<0.05）。生物信息学分析显示Linc00176可能通过靶向调控miR-338-3p的表达，进一步影响SMO蛋白的表达。双荧光素酶报告基因检测显示转染Linc00176-WT的miR-338-3p细胞（0.33±0.08）的相对荧光素酶活性显著低于miRNA-Control组细胞（1.25±0.16），差异有统计学意义（P<0.05）。CCK-8和异种移植瘤实验显示shRNA-Linc00176和miR-338-3p组细胞的增殖活力和成瘤能力与其各自的对照组相比显著下降，差异均有统计学意义（P<0.05）。Transwell结果表明与shRNA-Control组（134.47±12.29）和miRNA-Control组（129.87±13.06）比较，shRNA-Linc00176组（53.17±8.74）和miR-338-3p组（44.61±8.02）细胞的迁移个数均显著下降，差异有统计学意义（P<0.05）。Western blot结果显示，shRNA-Linc00176和miR-338-3p组细胞中SMO的蛋白表达水平较shRNA-Control和miRNA-Control组均显著降低，差异有统计学意义（P<0.05）。［结论］ Linc00176可能通过靶向调控miR-338-3p/SMO的表达促进HCC细胞的增殖和迁移。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effect of long-chain non-coding RNA Linc00176 targeting miR-338-3p/SMO on the proliferation and migration of hepatocellular carcinoma(HCC) cells. ［Methods］ Ninety-six specimens of HCC and adjacent tissues from patients with hepatocellular carcinoma were collected. Fluorescence quantitative PCR was used to detect the expression of Linc00176 and miR-338-3p in HCC and adjacent tissues. The shRNA-Control，shRNA-Linc00176，miRNA-Control and miR-338-3p lentiviral hepatoma cell lines were constructed. The targeting relationship between Linc00176 and miR-338-3p was analyzed by bioinformatics and dual luciferase reporter assay. The proliferation，migration，and the expression of SMO in each group were detected by CCK-8，xenograft tumor experiment and Western blot，respectively. ［Results］ The mRNA relative expression of Linc00176 in HCC tissues(1.94±0.21) was significantly higher than that in adjacent tissues(0.57±0.10)，and the mRNA relative expression of miR-338-3p in HCC tissues(0.35±0.08) was significantly lower than that in adjacent tissues(1.62±0.15)(P<0.05). Bioinformatics showed that Linc00176 may further affect the expression of SMO by targeting miR-338-3p. Dual luciferase reporter assay showed that the relative luciferase activity of miR-338-3p cells(0.33±0.08) transfected with Linc00176-WT was significantly lower than that of miRNA-Control cells(1.25±0.16)(P<0.05). CCK-8 and xenograft tumor experiments showed that the proliferation and tumorigenic ability of shRNA-Linc00176 and miR-338-3p cells were significantly decreased compared with their respective control groups(P<0.05). Transwell showed that compared with shRNA-Control(134.47±12.29) group and miRNA-Control(129.87±13.06) group，the number of migration in shRNA-Linc00176(53.17±8.74) group and miR-338-3p(44.61±8.02) group were decreased significantly(P<0.05). Western blot showed that the expressions of SMO in shRNA-Linc00176 group and miR-338-3p group were significantly lower than those in shRNA-Control group and miRNA-Control group(P<0.05). ［Conclusion］ Linc00176 may promote proliferation and migration of HCC cells by targeting the miR-338-3p/SMO axis. |
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