[1]魏 英,王小林.肝癌组织中lncRNA-PRR34-AS1的表达特性及其对肝癌细胞增殖、迁移的影响和潜在分子机制[J].现代检验医学杂志,2021,36(06):41-46.[doi:10.3969/j.issn.1671-7414.2021.06.008]
 WEI Ying,WANG Xiao-lin.Expression Characteristics of lncRNA-PRR34-AS1 in Liver Cancer Tissues and Its Effects on Proliferation and Migration of Liver Cancer Cells and Potential Molecular Mechanism[J].Journal of Modern Laboratory Medicine,2021,36(06):41-46.[doi:10.3969/j.issn.1671-7414.2021.06.008]
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肝癌组织中lncRNA-PRR34-AS1的表达特性及其对肝癌细胞增殖、迁移的影响和潜在分子机制()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第36卷
期数:
2021年06期
页码:
41-46
栏目:
论 著
出版日期:
2021-12-15

文章信息/Info

Title:
Expression Characteristics of lncRNA-PRR34-AS1 in Liver Cancer Tissues and Its Effects on Proliferation and Migration of Liver Cancer Cells and Potential Molecular Mechanism
文章编号:
1671-7414(2021)06-041-07
作者:
魏 英王小林
(榆林市第二医院a.肿瘤放疗科;b.普外科,陕西榆林 719000)
Author(s):
WEI Ying WANG Xiao-lin
(a. Department of Oncology Radiotherapy; b. Department of General Surgery, the Second Hospital of Yulin City, Shaanxi Yulin 719000,China)
关键词:
肝癌长链非编码RNA-PRR34-AS1JAK1增殖迁移
分类号:
R735.7;R730.43
DOI:
10.3969/j.issn.1671-7414.2021.06.008
文献标志码:
A
摘要:
目的 分析长链非编码RNA-PRR34-AS1在肝癌组织中的表达特性,探究其对肝癌细胞增殖、迁移的影响及潜在分子作用机制。方法 收集2020年6月~12月于榆林市第二医院行手术治疗的30例肝癌患者癌组织及其对应癌旁正常组织标本,采用实时荧光定量PCR实验(RT-qPCR)检测组织中LncRNA-PRR34-AS1相对表达水平。通过转染siRNA介导敲低PRR34-AS1基因表达,利用细胞增殖实验和细胞迁移实验验证PRR34-AS1对肝癌细胞增殖、迁移的影响;通过生物信息学数据库网站预测PRR34-AS1与JAK1结合的基因位点,双荧光素酶报告基因实验验证两者靶向关系,探究其在肝癌发生发展中的调控作用。结果 30例临床肝癌组织中PRR34-AS1表达显著高于癌旁正常组织(5.714±0.612 vs 2.981±0.572),差异有统计学意义(t=17.870,P=0.000),PRR34-AS1具有高表达预后差的临床特征。敲低PRR34-AS1后,转染24,48和72 h时siRNA-PRR34-AS1#1组和siRNA-PRR34-AS1#2组细胞增殖率较对照组明显降低,差异均有统计学意义(F=83.440~89.297,均P<0.001);siRNA-PRR34-AS1#1组(38.451±4.263)和siRNA-PRR34-AS1#2组(42.106±3.512)细胞愈合迁移速率较对照组(83.247±6.205)显著减慢,差异均有统计学意义(F=83.357,P<0.001)。JAK1是PRR34-AS1的靶基因,siRNA-PRR34-AS1#1组(0.453±0.019)和siRNA-PRR34-AS1#2组(0.476±0.022)细胞中JAK1相对表达较对照组相比(1.002±0.003)明显降低,差异均有统计学意义(F=716.287,P<0.001)。转染敲低JAK1表达后,各转染时间点siRNA-JAK1#1组和siRNA-JAK1#2组细胞增殖速率较对照组明显减缓,差异均有统计学意义(F=45.465~76.548,均P<0.05)。30组临床肝癌组织中JAK1表达显著高于癌旁正常组织(5.963±1.214 vs 4.052±0.876),差异有统计学意义(t=6.992,P=0.000),PRR34-AS1与JAK1表达呈显著正相关(r=0.907,P<0.05)。在敲低PRR34-AS1表达的肝癌细胞中回补JAK1后,肝癌细胞增殖、迁移速率又回归到正常水平。结论 肝癌中PRR34-AS1显著高表达,其可能通过正向调控JAK1表达影响JAK/STAT信号通路,进而调控促进肝癌细胞的增殖和迁移。
Abstract:
Objective  To explore its effect on the proliferation and migration of hepatocallular carcinoma cell and its potential molecular mechanism. by analyzing the expression characteristics of long non-coding RNA-PRR34-AS1 in hepatocellular carcinoma. Methods The relative expression levels of lncRNA-PRR34-AS1 in the tissues of 30 patients with liver cancer who underwent surgical treatment in the Second Hospital of Yulin City from June 2020 to December 2020 were detected by real-time fluorescence quantitative PCR (RT-qPCR). The expression of PRR34-AS1 gene was knocked down by siRNA transfection, and the effect of PRR34-AS1 on the proliferation and migration of HCC cells was verified by cell proliferation assay and cell migration assay. The binding loci of PRR34-AS1 and JAK1 were predicted through bioinformatics database website, and the targeting relationship between PRR34-AS1 and JAK1 was verified by double luciferase reporter gene assay to explore the regulatory role of PRR34-AS1 in the occurrence and development of liver cancer. Results The expression of PRR34-AS1 in 30 clinical HCC tissues was significantly higher than that in adjacent normal tissues (5.714±0.612 vs 2.981±0.572), the difference was statistically significant(t=17.870, P=0.000), and PRR34-AS1 was characterized by high expression and poor prognosis. After knockdown of PRR34-AS1, the cell proliferation rate of SiRNA-PRR34-AS1#1 group and SiRNA -PRR34-AS1#2 group was significantly lower than that of control group at 24, 48 and 72 h after transfection, the differences were statistically significant(F=83.440~89.297, P<0.001). Cell healing migration rate in SiRNA-PRR34-AS1#1 group (38.451±4.263) and SiRNA-PRR34-AS1#2 group (42.106±3.512) was significantly slower than that in control group (83.247±6.205),the difference was statistically significant (F=83.357, P< 0.001). JAK1 was a target gene for PRR34-AS1, and the relative expression of JAK1 in siRNA-PRR34-AS1#1 group (0.453±0.019) and siRNA-PRR34-AS1#2 group (0.476±0.022) was significantly lower than that in the control group (1.002±0.003), the difference was statistically significant (F=716.287, P < 0.001).After knockdown of JAK1 expression, the cell proliferation rate in SiRNA-JAK1#1 and SiRNA-JAK1#2 groups was significantly slower than that in the control group at each transfection time point, the difference were statistically significant(F=45.465~76.548, P<0.05). The expression of JAK1 in the 30 groups was significantly higher than that in the adjacent normal tissues (5.963±1.214 vs 4.052±0.876), the difference was statistically significant(t=6.992, P=0.000), and there was a significant positive correlation between PRR34-AS1 and JAK1 (r=0.907, P<0.05). After JAK1 was supplemented in HCC cells with PRR34-AS1 knockdown, the proliferation and migration rate of HCC cells returned to normal level. Conclusion PRR34-AS1 was significantly overexpressed in HCC, which may affect the JAK/STAT signaling pathway through the positive regulation of JAK1 expression, thereby regulating and promoting the proliferation and migration of HCC cells.

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备注/Memo

备注/Memo:
基金项目:陕西省卫生健康委员会科研项目(2019D106)。作者简介: 魏英(1978-),女,硕士,副主任医师,研究方向:消化道肿瘤规范化、个体化多学科综合治疗及相关转化研究,E-mail:ddcsfge@163.com。通讯作者:王小林(1979-),男,硕士,副主任医师,研究方向:擅长胃肠道肿瘤的传统及微创手术以及综合治疗。
更新日期/Last Update: 1900-01-01