·论著·

LncRNA-PVT1对人胰腺癌细胞株HPAF-Ⅱ增殖和凋亡的影响*

彭娟菲#黄凤婷庄燕妍陈文颖张世能&

中山大学孙逸仙纪念医院消化内科(510120)

#Email: pengjfei0305@163.com

&本文通信作者，Email: shinengz@163.net

背景：近年来，长非编码RNA(lncRNAs)在肿瘤发生、发展中的作用日益受到重视。研究发现lncRNA-PVT1在多种人类恶性肿瘤中表达上调并发挥促癌作用。目的：探讨PVT1在人胰腺癌细胞中的表达及其对HPAF-Ⅱ细胞增殖和凋亡的影响。方法：以脂质体转染技术将siRNA-PVT1转染入HPAF-Ⅱ细胞；以real-time PCR检测PVT1 mRNA表达，MTS实验和平板克隆形成实验检测细胞增殖能力，流式细胞分析检测细胞周期和细胞凋亡，蛋白质印迹法检测凋亡相关蛋白和原癌基因蛋白c-Myc表达。结果：以人永生化正常胰腺导管上皮细胞株H6c7作为对照，各人胰腺癌细胞株中以HPAF-Ⅱ细胞PVT1 mRNA表达升高最为明显。与转染阴性对照siRNA和未予转染siRNA的细胞相比，转染siRNA-PVT1的HPAF-Ⅱ细胞PVT1 mRNA表达显著降低，细胞增殖能力减弱，发生细胞周期G1期阻滞并出现凋亡峰，细胞中凋亡相关蛋白cleaved-caspase-3、cleaved-PARP表达上调，Bcl-2/Bax比值降低，c-Myc蛋白表达下调。结论：LncRNA-PVT1在人胰腺癌细胞株HPAF-Ⅱ中呈高表达，其可能通过调控c-Myc表达影响HPAF-Ⅱ细胞的增殖和凋亡。

关键词胰腺肿瘤；长非编码RNA；PVT1；原癌基因蛋白质c-myc；细胞增殖；细胞凋亡

Effect of LncRNA-PVT1 on Proliferation and Apoptosis of Human Pancreatic Cancer Cell Line HPAF-Ⅱ

PENGJuanfei,HUANGFengting,ZHUANGYanyan,CHENWenying,ZHANGShineng.

DepartmentofGastroenterology,SunYat-senMemorialHospital,SunYat-senUniversity,Guangzhou(510120)

Correspondence to: ZHANG Shineng, Email: shinengz@163.net

Background: Recent studies have shown that long non-coding RNAs (lncRNAs) play important roles in carcinogenesis and cancer biology and the related context has attracted more and more attentions. PVT1, which encodes a lncRNA, is reported to be up-regulated and exhibit pro-oncogenic activity in a wide variety of human cancers. Aims: To investigate the expression of PVT1 in human pancreatic cancer cells and its effect on proliferation and apoptosis of HPAF-Ⅱ cells. Methods: One target siRNA against PVT1 was synthesized and transfected into HPAF-Ⅱ cells by using lipofactamine technique. PVT1 mRNA expression was detected by real-time PCR; capability of cell proliferation was examined by MTS and colony formation assays; cell cycle progression and apoptosis were measured by flow cytometry; and Western blotting was performed to determine the expressions of apoptosis-related proteins and proto-oncogene protein c-Myc. Results: The mRNA expression of PVT1 in several human pancreatic cancer cell lines, especially HPAF-Ⅱ cells was significantly higher than that in H6c7, a human immortalization normal pancreatic ductal epithelial cell line. Compared with HPAF-Ⅱ cells transfected with negative control siRNA or without transfection, silencing of PVT1 by siRNA-PVT1 resulted in remarkable reduction in cell proliferation, cell cycle G1 phase arrest, and notable apoptosis; meanwhile, the expressions of apoptosis-related proteins (cleaved-caspase-3 and cleaved-PARP) were up-regulated, the ratio for Bcl-2/Bax was decreased, and the expression of c-Myc protein was down-regulated. Conclusions: LncRNA-PVT1 is highly expressed in human pancreatic cancer cell line HPAF-Ⅱ. It may affect the proliferation and apoptosis of HPAF-Ⅱ cells partially through regulating c-Myc expression.

Key wordsPancreatic Neoplasms;Long Non-Coding RNA;PVT1;Proto-Oncogene Proteins c-myc;

Cell Proliferation;Apoptosis

胰腺癌早期诊断率低，根治性手术率低，预后极差，死亡率高[1]，探讨胰腺癌的发生、发展机制有利于提高其临床诊治水平。近年来，长非编码RNA(long non-coding RNAs, lncRNAs)在肿瘤发生、发展中的作用日益受到重视。LncRNAs是一类转录本长度超过200 nt 的RNA分子，不编码蛋白质，但可通过表观遗传学以及转录和转录后调控层面调控基因表达[2]。……