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Molecular and Cellular Biochemistry

, Volume 370, Issue 1–2, pp 127–139 | Cite as

Silencing expression of ribosomal protein L26 and L29 by RNA interfering inhibits proliferation of human pancreatic cancer PANC-1 cells

  • Chaodong Li
  • Mei Ge
  • Yu Yin
  • Minyu Luo
  • Daijie ChenEmail author
Article

Abstract

Oncogenic KRAS, an important target for antitumor therapy, contributes to pancreatic cancer tumorigenesis, progression and maintenance. However, intracellular compensation regulation can help cells to resist the targeted therapy. Gene knockdown method such as RNAi may help to understand this intracellular regulatory system and discover novel therapeutic approach. In this study, two stable transfected cell lines were constructed through lentivirus-mediated shRNA targeting KRAS of PANC-1 cells, to investigate cell response to the knockdown of KRAS. Human whole genome microarray was then used to compare the gene expression profile. As a result, ribosomal proteins L26 and L29 (RPL26 and RPL29) were dramatically upregulated by KRAS-shRNA specifically. To identify whether RPL26 or RPL29 was critical for PANC-1 cells, siRNAs against RPL26 and RPL29 were designed and transfected in vitro. The results showed that knockdown of RPL26 or RPL29 expression significantly suppressed cell proliferation, induced cell arrest at G0/G1 phase and enhanced cell apoptosis. Reactive oxygen species (ROS) assay indicated that silencing of RPL26 or RPL29 markedly decreased the intracellular ROS generation. Our findings imply that siRNA interference against RPL26 and RPL29 is of potential value for intervention of pancreatic cancer.

Keywords

KRAS Lentivirus Microarray Pancreatic cancer RNA interference RPL26/RPL29 

Abbreviations

RNAi

RNA interference

shRNA

Short hairpin RNA

siRNA

Small interfering RNA

ROS

Reactive oxygen species

PC

Pancreatic cancer

FBS

Fetal bovine serum

ACDU

Automated cell deposition unit

FCM

Flow cytometry

NC

Negative control

qPCR

Quantitative real-time PCR

FITC

Fluorescein isothiocyanate

PI

Propidium iodide

PBS

Phosphate-buffered saline

DCFH-DA

2,7-Dichlorfluorescein-diacetate

DCF

2,7-Dichlorfluorescein

MTT

3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

DMSO

Dimethyl sulfoxide

GO

Gene Ontology

PVDF

Polyvinylidene fluoride

Notes

Acknowledgments

We are grateful to Dr Jianzhong Xi (Peking University, Department of Biomedical Engineering College of Engineering, Beijing, China) for lentiviruses production and critical reading of the manuscript. National New Drug Research and Development Project (Grant 2010ZX09401-403) supported this work.

Supplementary material

11010_2012_1404_MOESM1_ESM.xls (44 kb)
Supplementary material 1 (XLS 44 kb)
11010_2012_1404_MOESM2_ESM.doc (310 kb)
Supplementary material 2 (DOC 311 kb)

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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Chaodong Li
    • 1
    • 2
  • Mei Ge
    • 2
    • 3
  • Yu Yin
    • 2
    • 3
  • Minyu Luo
    • 2
  • Daijie Chen
    • 1
    Email author
  1. 1.School of BiotechnologyEast China University of Science and TechnologyShanghaiPeople’s Republic of China
  2. 2.Shanghai Laiyi Center for Biopharmaceutical R&DShanghaiPeople’s Republic of China
  3. 3.Shanghai Jiao Tong UniversityShanghaiPeople’s Republic of China

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