长链非编码RNA TSIX通过靶向miR-384调控人胰腺癌细胞增殖的研究

doi:10.12092/j.issn.1009-2501.2018.06.004

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (6): 621-626.doi: 10.12092/j.issn.1009-2501.2018.06.004

长链非编码RNA TSIX通过靶向miR-384调控人胰腺癌细胞增殖的研究

沈彬彬¹，周俊¹，黎亮¹，陆宁¹，姚明²

¹嘉兴市第一医院普外科，嘉兴 314001，浙江；²嘉兴市第一医院疼痛科，嘉兴 314001，浙江

收稿日期:2018-03-29 修回日期:2018-04-24 出版日期:2018-06-26 发布日期:2018-06-19
通讯作者: 周俊，男，研究生学历，主任医师，研究方向：lncRNA在胰腺癌中的作用机制研究。 Tel：13705739702Email：jxyyshenbinbin@163.com
作者简介:沈彬彬，女，本科，副主任医师，研究方向：lncRNA在胰腺癌中的作用机制研究。 Tel：13738268100Email：1425126923@qq.com
基金资助:
浙江省自然科学基金（LY16H090016）

Long non-coding RNA TSIX regulates the proliferation of human pancreatic cancer cells by targeting miR-384

SHEN Binbin¹, ZHOU Jun ¹, LI Liang ¹, LU Ning¹, YAO Ming ²

¹Department of General Surgery, the First Hospital of Jiaxing, Jiaxing 314001, Zhejiang, China; ²Department of Pain, the First Hospital of Jiaxing, Jiaxing 314001, Zhejiang, China

Received:2018-03-29 Revised:2018-04-24 Online:2018-06-26 Published:2018-06-19

摘要/Abstract

摘要：

目的：观察长链非编码RNA（lncRNA）TSIX在胰腺癌组织及细胞中的表达，并探讨其对细胞增殖的影响及可能的机制。方法：采用qPCR 法检测 TSIX 在5种胰腺癌细胞株（BxPC-3、CAPAN-1、AsPC-1、 CFPAC-1和SW1990）、人正常胰腺导管上皮细胞HPDE6-C7和7例胰腺癌组织及其癌旁组织中的差异性表达。生物信息学预测并分析与TSIX互补配对的miRNA及下游基因。用携带TSIX siRNA基因的重组慢病毒颗粒（LV-siRNA）感染SW1990细胞作为实验组，以阴性对照慢病毒颗粒（LV-NC）作为对照组。通过qPCR检测低表达TSIX对miR-384和Gab2 mRNA表达的影响，通过Western blot检测Gab2和PI3K/AKT通路蛋白的表达，通过流式细胞术、MTT比色法、集落形成实验检测低表达TSIX对SW1990细胞周期、活力及增殖的影响。结果：与胰腺导管上皮细胞HPDE6-C7相比，TSIX在胰腺癌细胞株中均呈高表达（P<0.01），其中在SW1990细胞中表达水平最高（P<0.01）。TSIX在胰腺癌组织中的表达水平高于其癌旁组织，差异有统计学意义（P<0.05）。SW1990细胞转染TSIX siRNA后，qPCR检测细胞中TSIX表达量明显降低（P<0.01），miR-384表达升高（P<0.01），Gab2的mRNA表达降低（P<0.01）；低表达TSIX能显著阻滞细胞周期、抑制细胞活力和集落形成能力（P<0.01）。结论：lncRNA TSIX在胰腺癌组织及细胞中高表达，低表达TSIX可抑制SW1990细胞的增殖，其机制可能为TSIX通过与miR384互补配对调控Gab2基因的表达。

关键词: 长链非编码RNA, 微小RNA, 胰腺肿瘤, 细胞增殖

Abstract:

AIM: To observe the expression levels of long non-coding RNA (LncRNA-TSIX) in the pancreatic cancer tissues and cell lines, and to explore its effect on cell proliferation and its possible mechanism. METHODS: qPCR was used to detect the expression of TSIX in 5 pancreatic cancer cell lines (BxPC-3, CAPAN-1, AsPC-1, CFPAC-1 and SW1990) and human normal pancreatic ductal epithelial cells HPDE6-C7, and 7 pancreatic cancer tissues with para-cancer tissues were also collected. Bioinformatics prediction and analysis of miRNAs and downstream genes paired with TSIX were performed. SW1990 cells were infected with recombinant lentiviral vector carrying TSIX siRNA gene (LV-siRNA) as experimental group and negative control lentivirus particles (LV-NC) as control group. The effect of TSIX low-expression on miR-384 and Gab2 mRNA expression was detected by qPCR, and the expression of Gab2 and PI3K/AKT pathway related protein was determined by Western blot. The effects of TSIX low-expression on the cell cycle, viability and colony formation ability of SW1990 cells were detected by flow cytometry, MTT assay and colony formation assay. RESULTS:Compared with HPDE6-C7 cells, TSIX was highly expressed in the pancreatic cancer cell lines (P<0.01). The expression of TSIX was the highest in the SW1990 cells (P<0.01). The expression level of TSIX in pancreatic cancer tissues was higher than that in its adjacent tissues (P<0.05). The expression of TSIX in the SW1990 cells transfected with TSIX siRNA was significantly decreased (P<0.01), the expression of miR384 was increased (P<0.01), and the mRNA expression of Gab2 was decreased (P<0.01). Low expression of TSIX significantly inhibited the cell cycle, and inhibited the cell viability and colony formation ability (P<0.01). CONCLUSION: LncRNA TSIX is overexpressed in the pancreatic cancer tissues and cell lines. Knockdown of TSIX expression inhibits the proliferation of SW1990 cells. The possible mechanism is that TSIX regulates the expression of Gab2 gene by combining with miR-384.

Key words: long non-coding RNA, microRNA, pancreatic tumor, cell proliferation

中图分类号:

R965.2

沈彬彬，周俊，黎亮，陆宁，姚明. 长链非编码RNA TSIX通过靶向miR-384调控人胰腺癌细胞增殖的研究[J]. 中国临床药理学与治疗学, 2018, 23(6): 621-626.

SHEN Binbin, ZHOU Jun, LI Liang, LU Ning, YAO Ming. Long non-coding RNA TSIX regulates the proliferation of human pancreatic cancer cells by targeting miR-384[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(6): 621-626.

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长链非编码RNA TSIX通过靶向miR-384调控人胰腺癌细胞增殖的研究

Long non-coding RNA TSIX regulates the proliferation of human pancreatic cancer cells by targeting miR-384

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