Effects of silencing PLCεgene by RNA interference on crocin-induced proliferation regulation of human bladder cancer cell line BIU-87

JI Hui-ying

doi:10.13406/j.cnki.cyxb.2008.05.018

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Effects of silencing PLCεgene by RNA interference on crocin-induced proliferation regulation of human bladder cancer cell line BIU-87

更新时间：2024-01-17

- Effects of silencing PLCεgene by RNA interference on crocin-induced proliferation regulation of human bladder cancer cell line BIU-87
- Journal of Chongqing Medical University Issue 5, Pages: 536-540(2008)
- 作者机构：
  
  1. 重庆医科大学检验系临床检验诊断学省部共建重点实验室
  2. 重庆医科大学基础医学院七年制
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2008.05.018
  CLC： R737.14
- Published：2008，
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[1]冀慧莹,吴绮思,郭永灿,吕纯芳,赵培,罗春丽.RNAi沉默PLCε对Crocin抑制人膀胱癌BIU-87细胞增殖作用调控的研究[J].重庆医科大学学报,2008(05):536-540.

JI Hui-ying. Effects of silencing PLCεgene by RNA interference on crocin-induced proliferation regulation of human bladder cancer cell line BIU-87. [J]. Journal of Chongqing Medical University (5):536-540(2008)
[1]冀慧莹,吴绮思,郭永灿,吕纯芳,赵培,罗春丽.RNAi沉默PLCε对Crocin抑制人膀胱癌BIU-87细胞增殖作用调控的研究[J].重庆医科大学学报,2008(05):536-540. DOI： 10.13406/j.cnki.cyxb.2008.05.018.

JI Hui-ying. Effects of silencing PLCεgene by RNA interference on crocin-induced proliferation regulation of human bladder cancer cell line BIU-87. [J]. Journal of Chongqing Medical University (5):536-540(2008) DOI： 10.13406/j.cnki.cyxb.2008.05.018.

摘要

目的:观察RNAi沉默膀胱癌BIU-87细胞磷脂酶C-ε（Phospholipase C epsilon

PLCε）表达后对藏花素（Crocin）抑制人膀胱癌BIU-87细胞增殖作用的影响。方法:将携有siRNA基因的真核表达质粒PGenesil-PLCε转染BIU-87细胞后

分为5组:未转染组、pGenesil-NP阴性质粒组、pGenesil-PLCε质粒组、Crocin组、pGenesil-PLCε联合Crocin组。MTT法观察不同处理组对BIU-87细胞的生长抑制作用。RT-PCR法检测转染后对PLCεmRNA表达的影响及各处理组PCNA（Proliferating cell nuclear antigen）、CyclinD1 mRNA表达。Western-blot检测PCNA、CyclinD1蛋白表达。结果:RNAi可抑制BIU-87细胞78.06%的PLCεmRNA表达。pGenesil-PLCε联合Crocin组可增强Crocin对BIU-87细胞的生长抑制作用

48h抑制率达45.30%

且与pGenesil-PLCε质粒组和Crocin组相比较有显著性差异（P<0.01）。RT-PCR和Western-blot结果显示与RNAi沉默PLCε组比较

RNAi PLCε联合Crocin组明显下调了PCNA、Cyclin D1基因和蛋白的表达（P<0.05）;较单独Crocin组比较

RNAi PLCε联合Crocin组显著降低了PCNA基因和蛋白的表达（P<0.05）

降低了Cyclin D1蛋白的表达（P<0.05）

Cyclin D1 mRNA表达变化两组间无显著性差异（P>0.05）。结论:沉默PLCε基因可增强Crocin对BIU-87细胞的抑制作用

其机制与进一步下调PCNA、CyclinD1的表达有关。

Abstract

Objective:To investigate the effects of Crocin on proliferation regulation after silencing PLCεgene expression by RNAi interference in human bladder cancer cells BIU-87.Methods:BIU-87 cells were transfected with plasmid pGenesil-PLCε.The experiment was designed as five groups to compared with each other

they were non-transfected cells as control group

pGenesil-NP negative plasmid transfected group

pGenesil-PLCεplasmid transfected group

Crocin group

pGenesil-PLCεplasmid transfected together with Crocin group.The influence on proliferation of every group was determined by methyl thiazolyl tetrazolium(MTT).The expression of PLCεmRNA of BIU-87 cells after transfection with the plasmid pGenesil-PLCεwas observed by RT-PCR.RT-PCR and Western-blot were used to detect cyclin D1 and PCNA expression. Results:78.06% mRMA of PLCεwas reduced clearly after transfected with plasmid pGenesil-PLCεin BIU-87 cells. pGenesil-PLCεplasmid transfected together with crocin group could enhance the inhibitive effect of Crocin on proliferation up to 45.30% at 48h

which was strikingly compared with the pGenesil-PLCεplasmid transfected group and the Crocin group (P<0.01).Both RT-PCR and western-blot indicated that the expression of PCNA

CyclinD1 both in mRNA and protein were markedly decreased in the RNAi together with Crocin group compared with the RNAi group(P<0.05).Compared with the Crocin group

The expression of PCNA both in mRNA and protein were reduced remarkably(P<0.05)

the expression of CyclinD1 protein was decreased clearly(P<0.05)in the RNAi together with Crocin group

but the expression of CyclinD1 mRNA had no significant difference between the two groups(P>0.05).Conclution:RNAi PLCεtogether with Crocin can enhance the inhibit effect of Crocin

they can further reduce PC NA

CyclinD1 expressionthe was the mechanism.

关键词

RNA干扰膀胱癌Crocin增殖

Keywords

RNAibladder cancerCrocinproliferation

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