Promoting effect of RNA-interfered pyruvate dehydrogenase kinase isozyme-1 on 5-fluorouracil-induced apoptosis of colon cancer cells

谢贛丰; 梁后杰; 童晶涛; 何金霞

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Promoting effect of RNA-interfered pyruvate dehydrogenase kinase isozyme-1 on 5-fluorouracil-induced apoptosis of colon cancer cells

更新时间：2026-01-09

- Promoting effect of RNA-interfered pyruvate dehydrogenase kinase isozyme-1 on 5-fluorouracil-induced apoptosis of colon cancer cells
- Journal of Army Medical University Vol. 34, Issue 1, Pages: 9-12(2012)
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  CLC： R394-33;R730.23;R735.35
- Published：2012
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谢贛丰,梁后杰,童晶涛,何金霞.RNA干扰PDK-1对5-FU诱导结肠癌细胞凋亡的促进作用[J].陆军军医大学学报(原第三军医大学学报),2012,34(01):9-12.

　Xie Ganfeng,Liang Houjie,Tong Jingtao,He Jinxia.Promoting effect of RNA-interfered pyruvate dehydrogenase kinase isozyme-1 on 5-fluorouracil-induced apoptosis of colon cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(01):9-12.
谢贛丰,梁后杰,童晶涛,何金霞.RNA干扰PDK-1对5-FU诱导结肠癌细胞凋亡的促进作用[J].陆军军医大学学报(原第三军医大学学报),2012,34(01):9-12. DOI：

　Xie Ganfeng,Liang Houjie,Tong Jingtao,He Jinxia.Promoting effect of RNA-interfered pyruvate dehydrogenase kinase isozyme-1 on 5-fluorouracil-induced apoptosis of colon cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(01):9-12. DOI：

摘要

目的探讨以丙酮酸脱氢酶激酶-1（pyruvate dehydrogenase kinase isozyme 1

PDK-1）为靶标的短发夹RNA（shRNA）与化疗药5-氟尿嘧啶（5-FU）联合应用对人结肠癌LS174T细胞的促凋亡作用。方法针对PDK-1序列设计3段干扰序列及1个阴性对照，分别构建pGenesil.1-PDK-1重组质粒表达载体及pGenesil.1-NC阴性对照载体，利用脂质体分别将质粒转染入LS174T，并用G418对细胞进行药物筛选，并通过Real-time PCR和Western blot检测有干扰序列的有效性。MTT实验检测5-FU对干扰组和对照组细胞生长、增殖的影响，流式细胞仪检测细胞凋亡率。结果 Real-time PCR和Western blot证实3个干扰组均抑制PDK-1表达，其中最大敲减效率超过70%。MTT实验结果显示，经5-FU同样处理48 h后，PDK-1干扰的LS174T细胞生长抑制率较对照组显著增高（P0.05）；流式细胞仪结果显示，干扰组细胞凋亡率较对照组增高（P0.05）。结论 PDK-1 shRNA能特异性下调LS174T细胞中PDK-1的表达，PDK-1干扰联合5-FU可促进结肠癌细胞化疗诱导凋亡。

Abstract

Objective To study the effect of pyruvate dehydrogenase kinase isozyme 1(PDK-1) on 5-fluorouracil (5-FU)-induced apoptosis of colon cancer cells. Methods Three interference sequences (interference group) and a negative control sequence of PDK-1 shRNA (control group) were designed to construct pGenesil-1 recombinant plasmid expression vector and pGenesil.1 negative control (pGenesil.1-NC) plasmid vector

respectively. The plasmid was transfected into LS174T cells using liposome and drugs were screened with G418. Effect of RNA interference on 5-FU-induced apoptosis of colon cancer cells was detected by real-time PCR and Western blotting

respectively. Effect of 5-FU on growth and apoptosis of colon cancer cells was detected by MTT assay and flow cytometry

respectively. Results Real-time PCR and Western blotting showed that the 3 interference sequences of PDK-1 shRNA inhibited the expression of PDK-1 in LS174T cells

with a maximum inhibitory rate of over 70%. MTT assay demonstrated that the growth inhibiting rate of LS174T cells interfered with PDK-1 was significantly higher in interference group than in control group 48 h after 5-FU was used (P0.05). Flow cytometry displayed that the apoptosis rate of colon cancer cells was significantly higher in interference group than in control group (P0.05). Conclusion PDK-1 shRNA can down-regulate the expression of PDK-1 in LS174T cells and PDK-1 interference in combination with 5-FU can promote the apoptosis of colon cancer cells.

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