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[长链非编码RNA-LINC01410在胰腺癌中的表达及其对胰腺癌细胞增殖和迁移的影响]

[The expression of long non-coding RNA-LINC01410 in pancreatic cancer and its effect on proliferation and migration of pancreatic cancer cells].

作者信息

Cai M, Xu L, Shen L, Zhang J

机构信息

Department of General Surgery, the First Hospital of Jiaxing, Jiaxing 314001, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2019 May 14;99(18):1406-1411. doi: 10.3760/cma.j.issn.0376-2491.2019.18.010.

Abstract

To explore the expression of long non-coding RNA-LINC01410 in the pancreatic cancer tissues, corresponding paracancerous tissues, human pancreatic cancer cell lines and normal pancreatic ductal epithelial cell line, and analyze the effect of LINC01410 on pancreatic cancer cell proliferation and migration. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression level of LINC01410 in 16 cases of pancreatic cancer tissue and its adjacent tissues. RT-qPCR was performed to analyze LINC01410 expression in the pancreatic cancer cell lines AsPC-1, CAPAN-1, SW1990, BxPC-3 and CFPAC-1, and human normal pancreatic ductal epithelial cell line HPDE6-C7. Transfection of interference plasmid (shRNA) in the pancreatic cancer cell line with the highest expression level of LINC01410 were used to knock-down the expression of LINC01410. CCK-8 assay, colony formation assay, and transwell chamber assay were performed to detect the proliferation and migration of pancreatic cancer cells. The complementary paired miRNAs and downstream genes of LINC01410 were predicted by bioinformatics. The expression of miRNA and downstream genes was detected by RT-qPCR, and the protein expression of downstream genes was determined by Western blot. The expression of LINC01410 in the pancreatic cancer tissues was significantly higher than that in the adjacent tissues [(3.46±0.32) vs (0.65±0.08), 0.01]. The expression levels of LINC01410 in the pancreatic cancer cell lines were significantly higher than that in the normal human pancreatic ductal epithelial cells (0.05). The expression of LINC01410 was highest in BxPC-3 cells (0.01). After knock-down of the LINC01410 expression in the pancreatic cancer cell line BxPC-3, the cell proliferation was significantly inhibited (0.05), and the cell migration ability was decreased (0.05). LINC01410 complementarily paired with miR-497-5p, and miR-497-5p complementarily paired with IFITM3. After inhibiting the expression of LINC01410, the expression of miR-497-5p was increased [(1.04±0.17) vs (5.79±0.43), 0.01], the mRNA expression of IFITM3 was decreased [(0.39±0.05) vs (1.00±0.03), 0.01], and the protein expression of IFITM3, CDK6, Cyclin D2, PCNA, Vimentin, and N-cadherin was decreased. The expression of LINC01410 was increased in pancreatic cancer tissues and cell lines. Down-regulation of LINC01410 expression inhibits the proliferation and migration of pancreatic cancer BxPC-3 cells, and its mechanism may be closely related to regulating the miR-497-5p and IFITM3 gene expression.

摘要

为探讨长链非编码RNA-LINC01410在胰腺癌组织、相应癌旁组织、人胰腺癌细胞系及正常胰腺导管上皮细胞系中的表达情况,并分析LINC01410对胰腺癌细胞增殖和迁移的影响。采用实时荧光定量PCR(RT-qPCR)检测16例胰腺癌组织及其癌旁组织中LINC01410的表达水平。运用RT-qPCR分析LINC01410在胰腺癌细胞系AsPC-1、CAPAN-1、SW1990、BxPC-3和CFPAC-1以及人正常胰腺导管上皮细胞系HPDE6-C7中的表达。在LINC01410表达水平最高的胰腺癌细胞系中,转染干扰质粒(shRNA)以敲低LINC01410的表达。采用CCK-8法、集落形成试验和Transwell小室试验检测胰腺癌细胞的增殖和迁移情况。通过生物信息学预测LINC01410的互补配对miRNA及其下游基因。采用RT-qPCR检测miRNA和下游基因的表达,通过蛋白质免疫印迹法检测下游基因的蛋白表达。LINC01410在胰腺癌组织中的表达显著高于癌旁组织[(3.46±0.32)对(0.65±0.08),P<0.01]。LINC01410在胰腺癌细胞系中的表达水平显著高于人正常胰腺导管上皮细胞(P<0.05)。LINC01410在BxPC-3细胞中的表达最高(P<0.01)。敲低胰腺癌BxPC-3细胞系中LINC01410的表达后,细胞增殖受到显著抑制(P<0.05),细胞迁移能力下降(P<0.05)。LINC01410与miR-497-5p互补配对,miR-497-5p与IFITM3互补配对。抑制LINC01410的表达后,miR-497-5p的表达升高[(1.04±0.17)对(5.79±0.43),P<0.01],IFITM3的mRNA表达降低[(0.39±0.05)对(1.00±0.03),P<0.01],IFITM3、CDK6、Cyclin D2、PCNA、波形蛋白和N-钙黏蛋白的蛋白表达均降低。LINC01410在胰腺癌组织和细胞系中表达上调。下调LINC01410的表达可抑制胰腺癌BxPC-3细胞的增殖和迁移,其机制可能与调控miR-497-5p和IFITM3基因表达密切相关。

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