UHRF1基因沉默对胰腺癌细胞增殖和凋亡的影响

严苹; 王清; 刘宇; 吴艳; 何帅; 陈霞

doi:10.3969/j.issn.1001-5256.2019.11.028

UHRF1基因沉默对胰腺癌细胞增殖和凋亡的影响

DOI: 10.3969/j.issn.1001-5256.2019.11.028

1. 西南医科大学附属医院消化内科
2. 乐山市人民医院肝胆外科

基金项目:

国家自然科学基金项目(090300021617)；

详细信息

中图分类号: R735.9
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出版历程
- 收稿日期: 2019-06-25
- 出版日期: 2019-11-20

Effect of UHRF1 gene silencing on the progression of pancreatic adenocarcinoma cells

Department of Gastroenterology,The Affiliated Hospital of Southwest Medical University

Research funding:

摘要

摘要: 目的探讨沉默UHRF1基因在胰腺癌细胞进展中的影响。方法选择2018年7月-2018年12月于乐山市人民医院肝胆外科行胰十二指肠切除术的30例胰腺癌患者,切取患者非坏死胰腺癌组织及相邻的癌旁组织为研究标本。采用半定量RT-PCR和Western Blot检测标本中UHRF1的mRNA和蛋白表达水平;随后构建靶向沉默UHRF1基因的短发夹RNA（sh-RNA）质粒表达载体,并采用慢病毒感染法转染胰腺癌细胞,MTT法检测细胞增殖; Western Blot法检测UHRF1、Bax和Bcl-2蛋白水平。计量资料两组间比较采用独立样本t检验;多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 UHRF1基因在胰腺癌组织中的表达水平显著高于癌旁组织（t=18. 131,P <0. 001）。与UHRF1组相比,UHRF1-sh RNA组癌细胞中UHRF1蛋白水平明显下调（t=3. 882,P=0. 023）,转染12 h及36 h后细胞增殖受到抑制（t值分别为4. 365、19. 042,P值分别为0. 005、<0. 001）; UHRF1-sh RNA组与癌旁组...
- 胰腺肿瘤 /
- 基因沉默 /
- UHRF1基因
Abstract: Objective To investigate the effect of UHRF1 gene silencing on the progression of pancreatic adenocarcinoma( PAAD) cells.Methods A total of 30 patients with PAAD who underwent pancreaticoduodenectomy in Department of Hepatobiliary Surgery,Leshan People's Hospital,from July to December,2018,were enrolled,and the non-necrotic PAAD tissue and the adjacent tissue were resected as samples.Semi-quantitative RT-PCR and Western blot were used to measure the mRNA and protein expression of UHRF1 in these samples. After the short-hairpin RNA( sh RNA) plasmid vectors were constructed for the targeted silencing of the UHRF1 gene and were transfected into PAAD cells by the lentivirus infection method,MTT assay was used to evaluate cell proliferation,and Western blot was used to measure the expression of UHRF1,Bax,and Bcl-2. The independent samples t-test was used for comparison of continuous data between two groups; a one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups. Results The expression of UHRF1 in PAAD tissue was significantly higher than that in adjacent tissue( t =18. 131,P <0. 001). Compared with the UHRF1 group,the UHRF1-sh RNA group had a significant reduction in the protein expression of UHRF1 in PAAD cells( t = 3. 882,P = 0. 023),with inhibited cell proliferation at 12 and 36 hours after transfection( t = 4. 365 and19. 042,P = 0. 005 and P < 0. 001). Compared with the adjacent tissue group and the UHRF1 group,the UHRF1-sh RNA group had a significant increase in the expression of the pro-apoptotic protein Bax( F = 862. 366,P < 0. 001) and a significant reduction in the expression of the anti-apoptotic protein Bcl-2( F = 170. 167,P < 0. 001),suggesting that UHRF1 gene silencing had a pro-apoptotic effect. Conclusion The expression of the UHRF1 gene is significantly upregulated in PAAD tissue,and UHRF1 gene silencing can inhibit the growth of PAAD tissue,possibly by inducing tumor cell apoptosis. UHRF1 may be a new molecular target for the treatment of PAAD.
- pancreatic neoplasms /
- gene silencing /
- UHRF1 gene

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参考文献(11)

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