长链非编码RNA LINC00922促进胰腺导管腺癌的增殖、迁移和侵袭
The long non-coding RNA LINC00922 promotes the proliferation, migration and invasion of pancreatic ductal adenocarcinoma cell line
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摘要:目的 探讨长链非编码RNA (long non-coding RNA, lncRNA) LINC00922在胰腺导管腺癌(pancreatic ductal adenocarcinoma, PDAC)组织及PDAC细胞系中的表达, 及对PDAC细胞系增殖、迁移和侵袭的影响。方法 利用illumina HiSeq X Ten高通量测序仪, 对2015年至2016年在复旦大学附属中山医院普通外科接受手术治疗的4例PDAC患者肿瘤标本及癌旁组织进行高通量测序, 建立lncRNA及mRNA表达谱, 并挑选其中在PDAC组织中显著高表达的lncRNA LINC00922做进一步研究。另选取2015年至2017年在复旦大学附属中山医院普通外科接受手术治疗的10例PDAC患者肿瘤标本及癌旁组织作为验证组, 采用实时定量反转录聚合酶链反应(RT-qPCR)技术验证LINC00922在PDAC中表达情况。通过CCK-8增殖实验、划痕实验及Transwell实验, 探讨LINC00922对PDAC细胞系BXPC-3增殖、迁移及侵袭的调控作用。采用配对t检验和独立样本t检验进行统计学分析。结果 高通量测序分析结果显示, PDAC组织中有470个差异表达lncRNA, 4 373个差异表达mRNA, 其中LINC00922在PDAC组织中显著高表达。在验证组10例PDAC及癌旁组织样本中, LINC00922在PDAC组织中显著高表达(P < 0.05)。CCK-8增殖实验、划痕实验以及Transwell侵袭实验分别提示敲低LINC00922表达的BxPC-3 PDAC细胞增殖活力、迁移能力以及侵袭能力均显著低于阴性对照组(P < 0.05)。结论 LINC00922在PDAC组织中显著高表达, 具有促进PDAC细胞增殖、迁移和侵袭的能力。Abstract:Objective To explore the expression of long non-coding RNA LINC00922 in pancreatic ductal adenocarcinoma (PDAC) tissues and PDAC cell lines and its function on proliferation and invasion of PDAC cell lines.Methods Appling the illumina HiSeq X Ten to high-throughput sequence four pairs of PDAC tissues and corresponding adjacent normal tissues during 2015-2016.Establishing the lncRNA and mRNA expression profile with the sequencing results.LINC00922 was highly expressed in PDAC tissues and was selected for further study.The expression of LINC00922 was verified by the RT-qPCR with another 10 pairs of pancreatic tissues and adjacent normal tissues.The function of LINC00922 on proliferation and invasion of pancreatic cancer cell lines was explored by CCK-8 cell proliferation assay, wound-healing assay and Transwell assay.Paired samples were analyzed by paired sample t test, or independent sample t test.Results Through high-throughput sequencing, 470 differentially expressed lncRNA and 4 373 differentially expressed mRNA were discovered in PDAC tissues.The lncRNA LINC00922 was over-expressed in PDAC.With another 10 paired pancreatic cancer tissues and adjacent normal tissues, the expression of LINC00922 was over-expressed in PDAC tissues (P < 0.05).CCK-8 cell proliferation assay showed that the proliferation activity of BxPC-3 was suppressed by down-expressing of LINC00922(P < 0.05).The wound healing assay showed that pancreatic cancer cells in LINC00922 knocking-down group was significantly more slowly closing to the scratch area than the negative control group (P < 0.05).The transwell assay showed that the number of migrating cells in LINC00922 knocking-down group was significantly lower than the negative control group (P < 0.05).Conclusions LINC00922 is over-expressed in PDAC and promotes the proliferation, migration and invasion of PDAC cells.
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