摘要
目的研究下调长链非编码RNA PVT1的表达对胰腺癌细胞增殖、凋亡及侵袭转移能力的影响。方法体外转染PVT1干扰序列si PVT1-1和si PVT1-2降低胰腺癌细胞系Bx PC-3 PVT1表达,同时转染阴性对照si PVT1-NC作为对照组。CCK-8实验检测细胞增殖,流式细胞术检测细胞凋亡,细胞划痕实验和Transwell实验检测细胞侵袭转移;qRT-PCR检测E-cadherin、N-cadherin、β-catenin、vimentin和PVT1 mRNA表达;Western blot检测凋亡相关蛋白(Bax、Bcl-2、caspase-3)和上皮间质转化相关蛋白(E-cadherin、N-cadherin、β-catenin、vimentin)表达。结果下调胰腺癌细胞Bx PC-3 PVT1表达能明显抑制细胞增殖、促进细胞凋亡和减少细胞侵袭转移数目(P<0.05);E-cadherin蛋白和分子水平升高,N-cadherin、β-catenin、vimentin蛋白和分子水平降低(P<0.05);Bax和caspase-3蛋白水平升高,而Bcl-2蛋白水平降低(P<0.05)。结论下调长链非编码RNA PVT1表达能抑制胰腺癌细胞增殖和侵袭转移能力,促进胰腺癌细胞凋亡,且能够逆转胰腺癌细胞上皮间质转化。
Objective To study the effects of down-regulated long non-coding RNA PVT1 expression on proliferation,apoptosis,migration and invasion of pancreatic cancer cells. Methods PVT1 interference sequences siPVT1-1 and siPVT1-2 were transfected into pancreatic cancer cell line BxPC-3 in vitro,and the siPVT1-NC was taken as negative control. The proliferation and apoptosis of BxPC-3 were detected by CCK-8 assay and Flow cytometry,respectively. Wound healing assay and Transwell assay were used to check ability of migration and invasion. The mRNA expression of E-cadherin,N-cadherin,β-catenin,vimentin and PVT1 were detected by qRT-PCR. Apoptosis related protein（ Bax,Bcl-2,caspase-3） and epithelial-mesenchymal transition related protein（ E-cadherin,N-cadherin,β-catenin,vimentin） were detected by Western blot. Results Down-regulated expression of pancreatic cancer cell line BxPC-3 reduced the ability of proliferation and promoted cell apoptosis（ P〈0. 05）. Thenumber of migrated and invased cells were significantly reduced when down-regulated the expression of PVT1（ P〈0. 05）. After transfection with siPVT1-1 and siPVT1-2,the molecular and protein level of E-cadherin increased,while N-cadherin,β-catenin and vimentin were reduced（ P〈0. 05）. The protein level of Bax and caspase-3 were increased,Bcl-2 was reduced（ P〈0. 05）. Conclusions Down-regulated expression of long non-coding RNA PVT1 can reduce cell proliferation and promote cell apoptosis,meanwhile,migration and invasion are also inhibited. Down-regulated expression of PVT1 can reverse epithelial-mesenchymal transition of pancreatic cancer cells.
出处
《基础医学与临床》
CSCD
2016年第1期73-79,共7页
Basic and Clinical Medicine
基金
镇江市科技计划(SH2014024)
江苏省卫生厅面上项目(H201434)
关键词
长链非编码RNA
PVT1
胰腺癌
上皮间质转化
long non-cading RNA
PVT1
pancreatic cancer
epithelial-mesenchymal transition
