Knockdown of MED19 by short hairpin RNA-mediated gene silencing inhibits pancreatic cancer cell proliferation.

Abstract

Abnormal gene transcription plays an important role in oncogenesis. In cancer cells, the improper activation of specific genes is usually ascribed to aberrant transcription machinery including transcription factors, RNA polymerase II, and Mediator complex. This study reports on short hairpin RNA (shRNA)-mediated gene silencing of MED19, a subunit of Mediator complex, and its effect on the growth of pancreatic cancer cells. RNA interference was performed by lentivirus shRNA system to specifically knockdown MED19 expression in Aspc-1 and Panc-1 cells. The knockdown efficiency of MED19 was confirmed by quantitative RT-PCR and western blot. The effect of MED19 shRNA on Aspc-1 and Panc-1 cell proliferation was evaluated by methylthiazoletetrazolium assay, BrdU incorporation assay, colony formation assay, and flow cytometry assay. This study shows that downregulation of MED19 remarkably reduced cancer cell proliferation and colony formation capacity in two pancreatic cancer cell lines. In addition, downregulated MED19 induced G1-phase cell cycle arrest and apoptosis. This study provides a potent role of MED19 in promoting pancreatic cancer growth and a possible drug target for cancer therapy.

• Authors: Xing-Hua Li, De-Ning Fang, Chai-Ming Zeng
• Journal: Cancer biotherapy & radiopharmaceuticals (Cancer Biother Radiopharm) Vol. 26 Issue 4 Pg. 495-501 (Aug 2011) ISSN: 1557-8852 [Electronic] United States
• PMID: 21834715 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• MED19 protein, human
• Mediator Complex
• RNA, Small Interfering

Topics

• Cell Growth Processes (genetics)
• Cell Line, Tumor
• Gene Expression
• Gene Knockdown Techniques
• Gene Silencing
• Genetic Therapy (methods)
• Humans
• Lentivirus (genetics, metabolism)
• Mediator Complex (antagonists & inhibitors, biosynthesis, genetics)
• Pancreatic Neoplasms (genetics, therapy)
• RNA, Small Interfering (administration & dosage, genetics)